Adeyemi 2009

11th Anniversary Annual International
Meeting of the Institute of Human Virology

Foreword from Robert C. Gallo, Founder and Director
For more than a decade, the Institute of Human Virology (IHV) at the
University of Maryland School of Medicine has pursued a mission of combined
research, treatment and prevention, in addition to a commitment and sense of
compassion towards a broader community that is not limited to the Baltimore
region.
IHV’s work knows no artificially drawn national boundaries. We are active at
laboratories and clinics, in the United States and abroad, via teamwork and in
partnership with scientists, clinicians, governments, and patients, among others.
Our measure of success continues to be an ability to take the results of never-
ending scientific research directly to the patient and in a manner that allows those
advances to have an effective global impact.
The Annual International Meeting of IHV has been a centerpiece of our efforts
to share with and learn from the broader scientific and clinical communities. The
Meeting is one of the world’s leading HIV/AIDS conferences, undoubtedly because
of the unparalleled quality of presentations and the level of expertise possessed by
presenters and participants. It is also because of the vision and dedication of
individuals like Hilary Koprowski and Stanley Prusiner who were instrumental in
organizing IHV’s inaugural meeting. I also want to acknowledge Jeff Meshulam of Profectus BioSciences, Inc., who has
contributed so much to so many of our meetings through the years, as well as Beth Peterson, whose tireless efforts now
reach fruition.
2008 marks an especially significant year as the anniversary of the finding of the HIV virus, while next year marks the
25th anniversary of the recognition of HIV as the cause of AIDS and the development of the first HIV blood test. These
significant milestones give us much reason to reflect upon what we did right, what has gone wrong and where we need to
go in the future.
Just recently, the field of HIV research suffered a major setback which in my view has too often been minimized. Interim
data from a large, expensive vaccine trial, the STEP trial, co-sponsored by the National Institute of Allergy and Infectious Disease
(NIAID) and Merck, showed that the vaccine employing an adenovirus vector with HIV genes had failed. Not only did the
vaccine offer no protection from HIV, it apparently increased the risk of infection in recipients who had previously been
exposed to adenoviruses similar to the vaccine vector. While failures are an unavoidable reality of grand scientific endeavors,
the fallout from the STEP trial presents an opportunity to re-evaluate the entire HIV vaccine development process.
In celebrating our 11th anniversary, we are honored to present the IHV Lifetime Achievement Award for Scientific
Contributions to Dr. Isaac P. Witz of Tel Aviv University. We will also be presenting an unprecedented two IHV Lifetime
Achievement Awards for Public Service to The Honorable Robert K. Gray, a Cabinet member in the Eisenhower Administration
and former worldwide Chairman of Hill & Knowlton, and Mr. John Evans, co-founder of C-SPAN and an internationally
recognized expert in the telecommunications industry.
We will also pause to commemorate the life of a good friend, Dr. Bob Ting – a research scientist, entrepreneur and
pioneer originating and helping to popularize the term ‘‘biotechnology.’’ Bob developed and produced the first FDA-
approved diagnostic test kits for HIV antibody confirmation and was the founding president and original chief executive of
Profectus Biosciences Inc.
I am grateful to all who have chosen to join us this year and am confident that this Meeting will write another
important chapter in the book of human retrovirology.
2008 International Meeting of the Institute of Human Virology SEPTEMBER 11-13, 2008 3
Copyright © 2009 Lippincott Williams & Wilkins. Unauthorized reproduction of this article is prohibited.
The Institute of Human Virology
www.ihv.org
Mission Statement
The Institute of Human Virology (IHV) is a world-class center of excellence focusing on chronic viral diseases and virally
linked cancers. IHV is dedicated to biomedical research leading to improved treatment and prevention of these diseases.
Our unique structure connects cohesive, multidisciplinary research and clinical programs so that new treatments are
streamlined from discovery to patients. IHV serves patients locally and the scientific community globally.
Robert C. Gallo
Director
Co-Director, Division of Basic Science and Vaccine Research
William A. Blattner
Associate Director
Director, Division of Epidemiology and Prevention
Robert R. Redfield
Associate Director
Director, Division of Clinical Care and Research
George Lewis
Co-Director, Division of Basic Science and Vaccine Research
Joseph L. Bryant
Director, Division of Animal Models
David Pauza
Assistant Director
Dave Wilkins
Chief Operating Officer
4 2008 International Meeting of the Institute of Human Virology SEPTEMBER 11-13, 2008
Board of Advisors
The Hon. Kathleen The Honorable Arthur Gajarsa Her Royal Highness Princess
Kennedy-Townsend United States Court of Appeals for Chulabhorn Mahidol
Chairperson the Federal Circuit Mahidol University
Washington, DC Bangkok, Thailand
Former Lieutenant Governor of
Maryland The Honorable Robert K. Gray Timothy Moynahan
Baltimore, Maryland Chairman, Gray and Company II Moynahan & Minella, LLC
Miami Beach, Florida Waterbury, Connecticut
The Honorable Sue Bailey
Assistant Secretary of Defense for Stewart Greenebaum Franco Nuschese
Health Affairs Greenebaum & Rose Associates, Inc. Georgetown Entertainment Group
Bethesda, MD Baltimore, MD Washington, DC
Fred Cannon William Hall Thomas Paese, Esquire
Senior Vice President for Government University College Dublin Buchanan Ingersoll and Rooney
Relations, BMI, Inc. Dublin, Ireland Harrisburg, Pennsylvania
New York, NY
William A. Haseltine Chris YH Tan
Robert Charrow Haseltine Foundation for Medical Asian Institute of Molecular and Cell
Greenberg-Traurig LLP Sciences and the Arts Biology
Washington, DC Washington, DC Vancouver, British Columbia
John P. Coale The Honorable Ernest F. Lenny Wilkens
Attorney at Law Hollings National Basketball Association Hall
Washington, DC Medical University of South Carolina of Fame Coach and Player
Charleston, SC Medina, Washington
Barbara J. Culliton
Science Journalist and Policy Richard E. Hug James Wyngaarden
Consultant HUG ENTERPRISES, INC. Former Director, National Institutes of
Washington, DC Baltimore, MD Health
Durham, North Carolina
The Honorable Elijah Toshiaki Inoue
Cummings Sanyo E&E America Company
United States House of Bensenville, IL
Representatives Ex-Officio Members
Baltimore, Maryland Mark Kaplan Robert C. Gallo
University of Michigan Medical Director, Institute of Human Virology
Lynda M. Dee Center
Attorney at Law, AIDS Activist Ann Arbor, Michigan William E. Kirwan
Baltimore, Maryland Chancellor
The Honorable Nancy Kopp University System of Maryland
Martin Delaney State Treasurer, Maryland State
Founding Director, Project Inform Government E. Albert Reece
San Francisco, California Annapolis, Maryland Dean
University of Maryland School of
The Honorable Sheila Dixon Hilary Koprowski Medicine
Mayor of Baltimore Thomas Jefferson University
Baltimore, Maryland Philadelphia, PA
John Evans Thomas Lynch
Evans Telecommunications Amarin Pharmaceutical, Ltd.
Key West, Florida Dublin, Ireland
Scientific Advisory Board
Hilary Koprowski, M.D. William Hall, M.D., Ph.D. Joseph Pagano, M.D.
Chair University College Dublin University of North Carolina School
Thomas Jefferson University Dublin, Ireland of Medicine
Philadelphia, Pennsylvania Mark Kaplan, M.D., FACP Chapel Hill, North Carolina
Edward A. Berger, Ph.D. University of Michigan
Kathleen Squires, M.D.
National Institutes of Health Medical Center
Ann Arbor, Michigan Thomas Jefferson University
Bethesda, Maryland
Philadelphia, Pennsylvania
Farley Cleghorn, M.D. Michel Klein, Ph.D.
Constella Group Canadian Network for Vaccines Mario Stevenson, Ph.D.
Washington, DC and Immunotherapeutics University of Massachusetts Medical
Myron S. Cohen, M.D. Toronto, Canada School
University of North Carolina School of Worcester, Massachusetts
Medicine
Myron Levine, M.D., D.T.P.H.
University of Maryland Sten Vermund, M.D., Ph.D.
Chapel Hill, North Carolina
School of Medicine Vanderbilt University Institute for
Max Essex, D.V.M., Ph.D. Baltimore, Maryland Global Health
Harvard School of Public Health Nashville, Tennessee
Cambridge, Massachusetts Erling C. J. Norrby, M.D., Ph.D.
Warner Greene, M.D., Ph.D. The Royal Swedish Academy of
Gladstone Institute of Virology and Sciences
Immunology Stockholm, Sweden
San Francisco, California
Communications and Press Policy

11th Annual International Meeting of the Institute of Human Virology
To enhance the exchange of information and communication among attendees of the Institute of Human Virology
Annual International Meeting, the following must be adhered to by all participants:
All comments at sessions are off-the-record and are not for attribution.
No coverage, reporting or publication of scientific data or presentations at the Institute of Human Virology Annual
Meeting is permitted without the consent of the presenter(s) and the program organizers.
One-on-one interviews with scientists and media may be arranged by contacting Nora Grannell, Director of Public
Relations and Marketing, Institute of Human Virology, ngrannell@ihv.umaryland.edu or (410) 706-1954.
Special Acknowledgements
The University of Maryland School of Medicine Institute of Human Virology expresses its gratitude to
the corporations, foundations, and US government institutions whose support makes possible the
11th Annual International Meeting of the Institute of Human Virology.
Benefactor
Fogarty International Center
The Bill & Melinda Gates Foundation
Underwriter
Office of AIDS Research
National Institutes of Health
Supporters
Gilead Sciences, Inc.
National Cancer Institute
OrthoClinicalDiagnostics, Inc.
Tibotec, Inc.
Contributors
Advanced Bioscience Laboratories, Inc.
Bristol-Myers Squibb
LabNow, Inc.
Partec Essential Healthcare
Sanofi-Aventis
Sanyo
Wyeth
Profectus Biosciences, Inc
11th Anniversary Annual International Meeting of the Institute of
Human Virology
ONLINE ABSTRACTS TITLE INDEX

To navigate to an abstract, simply click on the title of the abstract.
Abstracts will be available online at www.jaids.com
Thursday, September 11, 2008
HIV Entry
100 3D structure of native HIV-1 gp120 trimers and mechanisms of cellular entry
Sriram Subramaniam, National Cancer Institute, Bethesda, MD
101 Chemokine Receptor CCR5 Mediates Resistance to West Nile Virus Infection in Mouse and Man
Carole Bewley, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD
102 Visualization of HIV-1 entry pathways in cells

Gregory B. Melikian, Institute of Human Virology, University of Maryland School of Medicine
103 Tracking Dynamic HIV-1 Escape from CCR5 Antagonist Therapy in vivo by Deep Sequencing
Athe Tsibris, Massachusetts General Hospital, Boston, MA
104 A V3-independent pathway to resistance to small molecule CCR5 inhibitors

Kleio Anastasopolou, Weill Cornell Medical College, New York, NY
105 sCD4-17b, an Engineered Bifunctional HIV-1 Neutralizing Protein: Potent Cross Clade Activity and Potential
Microbicide Use
Laurel Lagenaur, National Institute of Allergy and Infectious Diseases, Bethesda, MD
HIV Infection Early Events

106 Early events in vaginal HIV transmission
Ronald Veazey, Tulane University
107 Escape from Neutralizing Antibody in Early Subtype C HIV-1 Infection

Cynthia A. Derdeyn, Emory University Vaccine Center, Altanta, GA
108 Indentification of Full-Length Transmitted HIV-1 Genomes Revelas Extraordinary Dynamics and Precise
Molecular Pathways of Early Virus Diversification, Adaptation and Immune Evasion
George Shaw, University of Alabama at Birmingham School of Medicine
109 Complexity of the Transmitted Virus and Compartmentalization In Its Seminal Source
Ronald Swanstrom, Center for AIDS Research University of North Carolina Lineberger Cancer Center
*Abstracts appear as provided by authors.
110 HIV permeation efficiency in human cervicovaginal mucus
Justin Hanes, Johns Hopkins University, Baltimore, MD
111 Reconstruction and Characterization of a Human Endogenous Retrovirus with the Original Sequence at the
Time of Integration
Reinhardt Kurth, Robert Koch-Institute, Berlin, Germany
112 hBD2 inhibits HIV via CCR6, a receptor expressed on memory, Dendritic and Th17 cells
Alfredo Garzino-Demo, Instittute of Human Virology, University of Maryland School of Medicine
113 The Anti-Herpetic Drug Acyclovir Supresses HIV-1 in Herpesvirus-Infected Human Tissues After Conversion
into a Nucleoside Reverse Transcriptase Inhibitor
Andrea Lisco, National Institute of Child Health and Human Development, Bethesda, MD
114 NK Cells Contribute to the Constitution of HIV Reservoirs in Dendritic Cells, Involvement of HMGB-1
Marie-Lise Gougeon, Institut Pasteur, Paris, France
115 HIV Vpu Complexes with bTrCP to Direct the Degradation of the Virus Release Inhibitor BST-2 (Tetherin)
Janet Douglas, Oregon Health and Science University
116 Molecular Studies of HIV-1 Rev Function

Lili Gu, Centre for Research in Infectious Diseases, Dublin, Ireland
117 CA-Dependent HIV-1 Nuclear Import Relies on Transportin-SR2

Vineet KewalRamani, National Cancer Institute, Frederick, MD
118 Self-Inactivation of HIV by its own RT/Rnase H

Karin Moellling, University of Zurich
119 HIV-1 Envelope gp120 Indicues a Stop Signal and Virological Synapse Formation in Non-infected CD4+
T Cells
Gaia Vasiliver-Shamis, New York University School of Medicine
120 Promoter-Targeted shRNA Driven by Retroviral Vector Achieves Long-term Repression of HIV-1 Replication
Makoto Yagamashi, University of Tokyo Institute of Medical Science
121 Plasmid DNA- and live viral vector-based vaccine approaches for the treatment and prevention of HIV
Infection
John Eldridge, Profectus Biosciences, Inc.
Progression/Anti-Progression Factors
122 Differential CD4 T cell subset depletion at mucosal sites in HIV infection
Daniel Douek, National Institute of Allergy and Infectious Diseases, Bethesda, MD
123 AIDS Restriction Genes-Genome Wide Association Study

Stephen O’Brien, National Cancer Institute, Frederick, MD
124 NK cells: good guys or bad guys?

Galit Alter, Massachusetts General Hospital, Boston, MA
125 Immunogenetic polymorphisms and their effects on AIDS progression
Xiaojiang Gao, National Cancer Institute, Frederick, MD
126 Host genetic determinants of response to HIV-1: A genome-wide perspective

Abstract not available at time of printing
David B. Goldstein, Duke Institute for Genome Sciences and Policy
127 HTLV-1 Infection of WE17/10 CD4+ Cell Line Leads to Progressive Alteration of Ca2+ Influx that Eventually
Results in Loss of CD7 Expression and Activation of an Antiapoptopic Pathway Involving AKT and BAD
Which Paves the Way for Malignant Transformation
Bassan Badram, University of Brussels, Belgium
128 Human Domain Antibodies Against HIV-1 as Exceptionally Potent Cross-Reactive Neutralizers
Weizao Chen, National Cancer Institute, Frederick, MD
129 Human Endogenous Retrovirus-K (HML-2) in the Plasma of People with Lymphoma and Breast Cancer
Rafael Contreras-Galindo, University of Michigan Medical Center, Ann Arbor, Michigan
130 Inhibition of HIV-1 Release by Cell Permeable Peptides

Sarah Daniels, National Institutes of Health, Bethesda, MD
131 The gp41-derived immunosuppressive (isu) peptide of HIV-1 modulates cytokine release and gene expression in
human immune cells
Joachim Denner, Robert Koch Institute, Berlin, Germany
132 Argotom-VAX: A Proposed Investigational HIV DNA Vaccine Composed of Selected Sequences of Clade B
Gag, Pol, Nef, Tat, Vif, and Env Linked to Lysosomal Membrane Associated Protein (LAMP)
William Hearl, Immunomic Therapeutics, Inc., Gaithersburg, MD
133 O-linked N-Acetylglucosaminylation Represses HIV-1 Replication and Sp-1 Mediated Trans- Activation of the
HIV-1 LTR
Ramona Jochmann, University of Erlangen, Germany
134 Cell-cycle Dependent HIV-1 Killing is Mediated Through the Viral Protease
Kyeongeun Lee, National Cancer Institute, Frederick, MD
135 Dramatic Enhancement of Uptake and Trafficking of HIV-1 Tat Protein via Modulation of Endocytosis
Pathways
Guan-Han Li, Johns Hopkins University School of Medicine, Baltimore, MD
136 Increased IL-15 Production is Associated with Higher Infection of Memory CD4 T Cells During Acute SIV
Infection
Joseph Mattapallil, Uniformed Services University of the Health Sciences, Bethesda, MD
137 Presence and Role of HLA-C in HIV-1 Infection

Andrea Matucci, University of Verona, Italy
138 Prevalence of Human Papillomavirus (HPV) Infection among HIV-Positive Women in the Pre- HAART and
HAART Era in a Nigeria Clinic
Olanrewaju Onigbogi, University College Hospital, Ibadan, Nigeria
139 HIV-2 Capsids Distinguishing High and Low Virus Load Patients in a West African Community Cohort
Clayton Onyango, MRC - Fajara Laboratories, Banjul, Gambia
140 A Retrospective Study of HIV-Exposed Infacts/Children in Kitwe, Zambia

Gilbert Siame, Zambia Rural Healthcare Outreach Services(ZARHOS), Kitwe, Zambia
141 Type C Coping and Alexithymia are Associated Differentially with Specific Immune Mechanisms (Interleukin-6
and Beta-Chemokine Production) Linked to HIV Progression
Lydia Temoshok, Institute of Human Virology, Baltimore, MD
142 HIV Frequently Elicits Mucosl and Plasma Env-Specific IgA with a Rapid Initial Decline in Acute Infection
Nicole Yates, Duke University Medical Center, Durham, NC
143 Broad Spectrum Neutralizing Antibodies Against HIV-1 Elicited by Immunizing with Fusion Complexes
Donato Zipeto, University of Verona, Italy
Friday, September 12, 2008

144 In vivo and ex-vivo proteomics for target discovery in cancer
Guiliano Elia, University College Dublin, Ireland
145 The V1/V2 Loop Region of Simian-Human Immunodeficiency Virus Envelope gp120 is the Major Determinant
of the Strain Specificity of the Neutralizing Antibody Response
Ronald C. Desrosiers, Harvard University, Cambridge, MA
146 a4b7: A Newly Discovered Receptor for HIV

Anthony Fauci, M.D., National Institute of Allergy and Infectious Diseases, Bethesda, MD
147 Hepatitis C and cancer

Michael Houghton, Epiphany Biosciences, San Francisco, CA
148 Towards a genetics of cancer resistance

George Klein, Karolinska Institutet, Stockholm, Sweden - In Honor of Lifetime Achievement Awardee Isaac Witz
HIV Malignancies
149 The Role of the Environment in EBV-Associated Lymphoid Malignancies
Eva Klein, Karolinska Institutet, Stockholm, Sweden
150 HIV and cancer: current trends and insights

Eric Engels, National Cancer Institute, Bethesda, MD
151 MicroRNAs and the biology of KSHV infection

Don Ganem, University of California, San Francisco
152 HIV-Associated Lymphomas. Focus on unusual lymphomas occurring specifically in HIV-infected patients
Antonino Carbone, Istituto Nazionale Tumori, Milano, Italy
Controlling Replication of Human Retroviruses
153 Insight into the molecular mechanisms of CIITA-mediated inhibition of HIV-1 and HTLV replication
Roberto Accolla, University of Insubria, Varese, Italy
154 Apobec3: A Modern Twist to A Classic Retroviral Mystery

Warner Greene, Gladstone Institute of Virology and Immunology, University of California San Francisco
155 Tetherin, An Interferon-alpha Induced Inhibitor of Retrovirus Release that is Antagonized by Vpu
Paul Bienasz, Aaron Diamond AIDS Research Center, New York, NY
156 Humanized Mouse Models for the In-Vivo Analysis of HIV Infection
Victor Garcia-Martinez, Southwestern Medical School, Dallas, TX
157 Vpr as a mediator of proteasomal degradation and other functions

Carlos de Noronha, Albany Medical College, Albany, NY
158 The SET complex acts as a barrier to autointegration of HIV-1

Judy Lieberman, Immune Disease Institute, Harvard Medical College
159 Finding Host Proteins Required for HIV Replication
Abraham Brass, Massachusetts General Hospital, Boston, MA
160 Polarization of M1 and M2 macrophages and HIV infection

Guido Poli, San Raffaele Scientific Institute, Milan, Italy
161 Peptide stabilization of gp120 confirmation: a novel vaccine candidate

Jonathan Gershoni, Tel Aviv University, Israel
Saturday, September 13, 2008

Clinical HIV Session
162 Transmission Networks of Drug Resistance Acquired in Primary/Early Stage HIV Infection
Mark Wainberg, McGill AIDS Centre, Lady Davis Institute, Jewish General Hospital
163 HIV Persistence in Patients on HAART: Re-evaluating Prospects for Eradication
John Bartlett, Center for Civilian Biodefense Strategies, Johns Hopkins University Bloomberg School of Public
Health
164 An update on raltegravir (Isentress)
Charles F. Farthing, Merck
165 Evaluation of Efficacy and Immune Recovery of Optimized Background Therapy (OBT) Plus Maraviroc
(MVC) vs Placebo (PBO) in Treatment Experienced (TE) Patients with only R5 HIV-1, Combined Analysis of
MOTIVATE 1 and 2
Randall Tressler, Pfizer
166 HIV Therapeutics in Resource Limited Settings – Status and Future Directions
Anthony Amoroso, Institute of Human Virology, University of Maryland School of Medicine
167 Update on Gilead Sciences Anti-HIV Development Programs
Tomas Cihlar, Gilead Sciences
168 Rapamycin enhances the anti-HIV activity of CCR5 antagonist Vicriviroc
Olga Latinovic, Institute of Human Virology
Vaccines, Immune Response and Innate Immunity
169 Why do so few Antibodies Neutralize HIV-1? Tests of the ‘‘Tolerance Hypothesis’’
Garnett Kelsoe, Department of Immunology, Duke University
170 Getting the Right Immune Response to HIV: Evaluation of Protective Immune Responses After Vaccination of
Rhesus Macaques
George Pavlakis, National Cancer Institute, Frederick, MD
171 Induction of Cross-clade Neutralizing Antibodies in Rabbits Using a DNA Prime/Protein Boost Immunization
Regimen
Susan Zolla-Pazner, New York University Medical Center
172 Prospects for an AIDS Vaccine: Can Effector-Memory T Cell Responses Contribute?
Louis T. Picker, Oregon Health and Science University
173 Human defensins - Small in Size, but Big in Functionality
Wuyuan Lu, Institute of Human Virology, University of Maryland School of Medicine
174 The C5 region of gp120; a therapeutic vaccine target?

Angus Dalgleish, St. George’s University of London
175 Mechanisms by which Synergistic Combinations of TLR Ligands Enhance T-Cell Responses to Vaccines
Jay Berzofsky, National Cancer Institute, Bethesda, MD
176 Vector-based Vaccines for Cancer Therapy

Jeffrey Schlom, National Cancer Institute, Bethesda, MD
177 Recruitment of High Avidity Antigen-Specific T Cells in Pancreatic Cancer Patients
Elizabeth M. Jaffee, M.D., Sidney Kimmel Cancer Center at Johns Hopkins
Preventative Vaccines
178 Development of Preventive HIV Vaccines in the US Military HIV Research Program
Nelson Michael, United States Military HIV Research Program, Rockville, MD
179 Rational Vaccine Design and the Development of an AIDS Vaccine

Gary Nabel, Vaccine Research Center, National Institutes of Health, Bethesda, MD
180 Continuing Clinical Trials for HIV Vaccine Research
Glenda Gray, Perinatal HIV Research Unit, University of the Witwatersrand
181 HIV-specific immune responses in healthy volunteers immunized with a multigene, multiclade HIV-1 DNA
vaccine and boosted with HIV-1 MVA in Sweden and Tanzania
Abstract not available at time of printing
Gunnel Biberfeld, Karolinska Institutet, Stockholm, Sweden
182 A Novel Model for In Vivo SIV Neutralization

Philip Johnson, Children’s Hospital of Philadelphia
183 Morphogenomic immune responsiveness to preventive/therapeutic vaccines

Luigi Buonaguro, Istituto Nazionale Tumori ‘‘Fond. G. Pascale,’’ Naples, Italy
184 Development of the broadly neutralizing human antibody m9 for anti-HIV prophylactics
Antony Dimitrov, Profectus Biosciences, Inc.
POSTER SESSION INDEX
185 Interaction between a domain in the fusion peptide proximal region (FPPR) of gp41 and the epitope domain in
the membrane proximal external region (MPER) increases binding of 2F5 to its epitope
Uwe Fiebig, Magdalena Eschricht, Mirco Schmolke, Reinhard Kurth, Joachim Denner, Robert Koch- Institute
186 Immunosuppressive human endogenous retrovirus K (HERV-K) is expressed in human villous and extravillous
cytotrophoblasts
Ulrike Kämmerer1, Ariane Germeyer2, Michaela Kapp1, Sven Stengel3, Kristina Büscher3, Reinhard Kurth3, and
Joachim Denner3; 1University Women’s Hospital, Wuerzburg, Germany; 2University of Heidelberg, Dept. of
Gynecological Endorinology and Reproductive Medicine, Heidelberg, Germany; and 3Robert Koch Institute, Berlin,
Germany
187 Autoresistance to X4 HIV Infection by soluble suppressor factors secreted from CD4+ T cells
F. Cocchi, A. DeVico, A. Garzino Demo and R. C. Gallo; Institute of Human Virology, University of aryland School
of Medicine
188 Targeted delivery of antiHIV siRNAs to T cells in vivo

Premlata Shankar, M.D., Department of Biomedical Sciences, Texas Tech University Health Sciences enter,
Paul L. Foster School of Medicine
189 Defective HIV-1 Proviral Genomes in Natural Viral Suppressors

L. M. Eyzaguirre, M. M. Sajadi, R. R. Redfield, W. A. Blattner and J. K. Carr; Institute of Human Virology, University
of Maryland School of Medicine
190 The Immune Response to HIV: Friend or Foe

M. Karen Newell1, Elizabeth Connick2, Evan Newell3, Haig Keledjian3, Monica Ord3, Robert Berliner3, Joshua
Cabrera1, Richard Tobin1, Cassie Pleasant1 and Lisa Villalobos-Menuey1; 1University of Colorado, Colorado
Springs; 2University of Colorado Health Sciences Center; and 3Viral Genetics, Inc., Asuza, California
191 Anti-FasL treatment preserves SIV-specific memory and slows progression to AIDS in rhesus macaques
Bhawna Poonia, Maria S. Salvato, C. David Pauza, Institute of Human Virology
192 Significant Relationship between INNO-LIAä HIV I/II Positivity Bands and the Immunological Status of HIV
Patients
Fernanda Leite, Fátima Oliveira and Luciana Pinho; Clinical Haematology Dept. - Hospital Geral Santo Antonio
193 The HIV-Positive Inpatient: Psychosocial Risks and Adherence Implications

Rebecca L. Wald, Stephen J. Synowski and Lydia R. Temoshok, Institute of Human Virology and Department of
Medicine, University of Maryland School of Medicine
193a Psychosocial Contributors to Antiretroviral Adherence: Stability and Change

Rebecca L. Wald, Stephen J. Synowski and Lydia R. Temoshok; Institute of Human Virology and Department of
Medicine, University of Maryland School of Medicine
194 Systolic Blood Pressure recovery following mental stress predicts immune dysregulation in persons with HIV
Stephen J. Synowski Ph.D., Institute of Human Virology, University of Maryland School of Medicine
195 Stimulation of reverse cholesterol transport potently suppresses HIV-1 replication

Michael Bukrinsky, The George Washington University
196 Monocyte-Dependent and –Independent Modulatory Effects of Vitamin D3 on X4 and R5 HIV-1 Replication in
IL-2 Stimulated PBMC
Massimo Alfano1, Chiara Rizzi1, Giuseppe Penna2, Luciano Adorini2 and Guido Poli1,3; 1San Raffaele Scientific
Institute; 2BioXell SpA; and 3Vita-Salute San Raffaele University, School of Medicine
197 Naturally Occurring C-terminally Truncated Isoform of STAT5 (STAT5D) Is a Transcriptional Repressor of
HIV-1 Expression. Characterization of the Platinum-Based STAT Inhibitor CPA-7
Giulia Della Chiara1, Andrea Crotti1, Heidi Kay2 and Guido Poli1 ; 1AIDS Immunopathogenesis Unit - San Raffaele
Scientific Institute, Milano, Italy; and 2College of Public Health, University of South Florida, Tampa, Florida
198 Modulation of CCR5 density with low doses of the transplant drug Rapamycin sensitizes Vicriviroc-resistant
R5 HIV-1
Alonso Heredia, Institute of Human Virology, Baltimore, MD
199 Risk factors for Virologic Failure and Adverse Reactions among Patients on Triple Antiretroviral therapy
Adedayo Adeyemi, Oluseyi Adesola and Oluyemisi Olaogun, Healthmatch International, Lagos, Nigeria
200 Design, Synthesis, Anti-HIV and Cytotoxicity of Novel Heterocyclic Compounds

Periyasamy Selvam, Amrita School of Pharmacy, Elmakkara, Kerala, India
200a Inhibition of HIV replication and integrase activity by isatin derivatives

Periyasamy Selvam, Amrita School of Pharmacy, Elmakkara, Kerala, India
201 Pathogenesis induced acidosis and hpercalceamia evident in HIV/AIDS disease

Abdulrazak Hamza Yahaya, HIV/Immunology Laboratory, Pathology Department Murtala Muhammad
Specialist Hospital, Kano, Nigeria
202 Retinopathy and enteropathy in HIV patients, a hypothetical concept beyond Virus and CD4 levels- a study
from Varanasi, North India
Dr. V. Satya Suresh Attili, Prof. Shyam Sundra, Prof. V. P. Singh and Prof. A. K. Gulati, Yashoda Hospitals,
Somajiguda, Hyderabad
203 Immunosuppression Level In HIV-1 –Infected Patients Doesn’t Affect the Serological Diagnosis of Hepatitis
C Virus
Fernanda Leite, Luciana Pinho; Clinical Haematology Dept- Hospital Geral Santo Antonio- Porto- Portugal
204 Serum albumin: Could it be an inexpensive and simple marker of immunosupression in HIV-infected
individuals?
Muthu Sundaram, YRG Centre for AIDS Research and Education (YRG CARE), Voluntary Health Services Campus,
Chennai, India
205 Increased IFN-alpha expression in circulating plasmacytoid dendritic cells of HIV-1 infected patients despite
selective loss
Clara Lehmann, M.D.1,3, Dirk Taubert, Ph.D.2, Jill M. Harper, Ph.D.3, Norma Jung, M.D.1, Pia Hartmann, M.D.1, Gerd
Fätkenheuer, M.D.1 and Fabio Romerio, PhD.3; 1First Department of Internal Medicine, University of Cologne,
Cologne, Germany; 2Department of Pharmacology, University of Cologne, Cologne, Germany; and 3Institute of
Human Virology, University of Maryland School of Medicine, Baltimore, MD
206 HIV-Induced Alterations in Vg2Vd2 T-cell Phenotype and Function Impact Mechanisms for Tumor Immunity
in AIDS
Jean-Saville Cummings, Institute of Human Virology, Baltimore, MD
207 Mechanisms regulating Vg2Vd2 tumor cell cytotoxicity: Roles for CD56 and T cell receptor
Kun Luo, Institute of Human Virology University of Maryland, Baltimore, MD
208 Altered cord blood gaT cell repertoire in Nigeria: possible impacts of environmental factors on neonatal
immunity
C. Cairo, Institute Of Human Virology, Baltimore, MD
209 TNF-a is an autocrine factor for Va2Va2 T cell

Haishan Li, Institute of Human Virology, Baltimore, MD
210 Peripheral blood mononuclear cells activated by HIV-VLPs in correlation with HIV-1 seropositivity status
L. Buonaguro1, M. L. Tornesello1, R. C. Gallo2, F. M. Marincola3, G. K. Lewis2 and F. M. Buonaguro1; 1Lab. of Viral
Oncogenesis and Immunotherapies and AIDS Reference Ceneter, Istituto Nazionale Tumori ‘‘Fond. G. Pascale’’,
Naples - Italy; 2Institute of Human Virology, Univ. of Maryland School of Medicine, Baltimore, MD, USA; and
3
Immunogenetics Section, Department of Transfusion Medicine, Clinical Center, NIH, Bethesda
211 Potent Orally Bioavailable HIV-1 Fusion Inhibitors Alter Env Conformation and Expose Conserved
Neutralization Epitopes
C. Finnegan, V. Dettmer, M. Bramah-Lawani, T. Nitz, P. Bullock, I. Burimski, M. Reddick, C. Matallana, C. Beaubien,
D. Stanley, J. Pettitt, G. Allaway and K. Salzwedel; Panacos Pharmaceuticals, Gaithersburg, MD
212 In vivo alteration of humoral responses to HIV-1 envelope gp120 by antibodies to the CD4-binding site of gp120
Maria Luisa Visciano, New York, NY
213 Dendritic cell-specific delivery of siRNA targeting SOCS1 enhances HIV-gag-specific CD8 T cell response
Sandesh Subramanya, Texas Tech University-Health Sciences Center, Department of Biomedical Sciences, El
Paso, TX
214 Increased expression of Suppressor of Cytokine Signaling-1 (SOCS-1) by HIV-1 transgenic rats: A mechanism
for dysregulated T helper-1 responses
William Reid, Institute of Human Virology, Baltimore, MD
215 The A1 subunit of Cholera toxin as an adjuvant for HIV DNA vaccines
Kenneth Bagley, Baltimore, MD
216 Genetics of Fc Gamma Receptors IIa and IIIa in Tanzania

Gustavo H. Kijak, Pharm.D., Ph.D.; U.S. Military HIV Research Program, Rockville, MD
217 HIV Knowledge and Willingness to Participate in New Preventive Technologies (NPT) Trials among a Nigerian
Refugee Population
O. Akinyemi, Department of Community Medicine, University College Hospital, Ibadan, Nigeria
218 Knowledge, attitudes and practices (KAP) of sexually active men towards circumcision as a preventive measure
against HIV infection in Kitwe district, Zambia
Sthembile Ndopu and Gilbert Siame, Zambia Rural Healthcare Outreach Services (ZARHOS), Kitwe, Zambia
219 Preventing HIV infection in developing countries: focus on gender and age as the determinants of the spread
Abdulrazak Hamza Yahaya, HIV/Immunology Laboratory, Murtala Mohammed Specialists Hospital Kano,
Nigeria
220 The Translational Laboratory Shared Services
Mariola Sadowska, Ph.D., Colette Burgess, Latreece Nance and Joseph Bryant, DVM, TLSS, UMGCC,
Baltimore, MD
221 Humoral Immunity against Conserved Epitopes of HIV-1 Envelope Protein Archived in Memory B cells in
Natural Viral Suppressors: Discordance with Plasma Antibodies
Yongjun Guan1, Mohammad Sajadi1, Anthony L. DeVico1, Christine Obriecht1, Robin Flinko1, Karla Godfrey1,
Timothy Fouts2, Ranajit Pal3, Robert Redfield1, Robert Gallo1 and George K. Lewis1; 1Divisions of Basic Sciences
and Vaccine and Clinical Sciences, Institute of Human Virology, University of Maryland School of Medicine,
Baltimore, MD; 2Perfectus BioSciences, TechCenter at University of Maryland Baltimore County, Baltimore, MD;
and 3Advanced BioScience Laboratories, Kensington, MD
222 Antibody 2G12 recognizes a glycopeptide epitope on HIV-1 gp120 envelope glycoprotein
Wei Huang, George K. Lewis and Lai-Xi Wang; Institute of Human Virology, University of Maryland School of
Medicine, Baltimore, MD
223 An animal model for non-hodgkin’s lymphoma of the central nervous system (NHL-CNS)
J. Bryant, H. Tran, M. Sadowska, E. Ateh and Y. Lunardi-Iskandar, Baltimore, MD
224 Persistent virological benefit in SIV-infected macaques upon therapeutic vaccination with DNA vectors by
in vivo constant-current electroporation
Barbara K. Felber1, A. Valentin2, A. von Gegerfelt2, M. Rosati2, V. Pate2, G. Miteloudis2, C. Alicea1, C. Bergamaschi2,
R. Jalah1, A. Kha3, R. Draghia-Akli3 and G.N. Pavlakis2; 1Human Retrovirus Pathogenesis Section, 2Human
Retrovirus Section, NCI, Frederick, MD; and 3VGX Pharmaceuticals, Inc., The Woodlands, TX
225 New Viral and Tuberculosis Therapeutics for the 21st Century
Roger J. Pomerantz, M.D., Johnson & Johnson Corporation
226 Glycosylation of gp41 of Simian Immunodeficiency Virus Shields Epitopes That Can Be Targets for
Neutralizing Antibodies
Eloı́sa Yuste, Jacqueline Bixby, Jeffrey Lifson, Shuji Sato and Ronald Desrosiers; Harvard Medical School
FOGARTY SCHOLARSHIP ABSTRACTS

227 Opportunity for Scale –Up; Mobile X-ray Technology- Strengthening TB Diagnosis in HIV+ve Patients;
ACTION Experience in Zaria, rural Northern Nigeria
U. I. Gebi, B. Musa, N. Alfred, A. Abimiku, P. Dakum, W. Blattner, E. Meshak, O. Obasanya, M. Gidado
and A. Clement; Institute of Human Virology, Nigeria
228 Psychological distress as a risk factor for non-adherence to Highly active anti-retroviral therapy
Etheldreda Nakimuli-Mpungu, Johns Hopkins School of Public Health
229 Baseline renal insufficiency and risk of death among HIV-infected adults on antiretroviral therapy
in Lusaka, Zambia
L. B. Mulenga1, G. Kruse1, S. Lakhi2, R. A. Cantrell1,3, S. E. Reid1,3, I. Zulu4,5, E. M. Stringer1,3, Z. Krishnasami3,
A. Mwinga4, M. S. Saag3, J. S. A. Stringer1,3, and B. H. Chi1,3; 1Center for Infectious Disease Research, Lusaka,
Zambia; 2University Teaching Hospital, Lusaka, Zambia; 3University of Alabama at Birmingham, AL; 4CDC,
Lusaka, Zambia; and 5University of Zambia School of Medicine, Lusaka, Zambia
231 Lopinavir/ritonavir-based Second Line Antiretroviral Treatment in children at National Pediatric Hospital,
Phnom Penh, Cambodia
S. Sam, V. Ung1, C. Huot1, C. Courpotin2, Eric Nerrienet3, David Pugatch4, Kenneth Mayer4 and Y. M. Chhour5 ; 1Child
Health Improvement Clinic, National Pediatric Hospital, Phnom Penh, Cambodia; 2French Red Cross, Phnom Penh,
Cambodia; 3Pasteur Institute, Phnom Penh, Cambodia; 4Brown University, Miriam Hospital, Providence, RI, USA; and
5
National Pediatric Hospital, Phnom Penh, Cambodia
232 Explosive Expansion of HIVand associated risk factors among Male and Hijra Sex Workers in Sindh, Pakistan
Arshad Altaf, HIV/AIDS Surveillance Project, Sindh AIDS Control Programme
233 Enhancement of HIV-1 Replication in Human Primary Cells by Macrophage Migration Inhibitory Factor in
Nigerian Africans
Dr. Busari, Olusegun Adesola, HIV Study Group, Federal Medical Centre
234 Neutralization efficiency and presence of anti-V3 antibodies in plasma of HIV-1 infected Northern Indians
Alok K. Choudhary1, Subhashree Dutta1, Naveet Wig2, A Biswas. 2, Raiees Andrabi1, Rajesh Kalra1, Rama Bhasin3,
Susan Zolla Pazner4, Suman Laal4, and Kalpana Luthra1; 1Department of Biochemistry; 2Department of
Medicine; 3Blood Bank, CN Centre, All India Institute of Medical Sciences, New Delhi, India; and 4Veterans Affairs
Medical Center, NYU School of Medicine New York, NY
235 Quality ART Scale Up Through Regionalization of Laboratory Services In Nigeria

C. Ezeaku, J. Farley, P. Dakum, T. Croxton, N. Constantin2, A. Abimiku and W. Blattner; Institute of Human
Virology Nigeria
236 T-Cell Receptor (TCR) Activation Mediates Efficient and Sustained HIV Transcriptional Elongation and
Initiation through Multiple Signal Pathways
Joseph F. Hokello; Department of Molecular Biology and Microbiology, CWRU School of Medicine
237 N-linked glycans on HIV-1 gp120 are critical determinants for the recognition of CD4 helper T cell epitopes
Hualin Li, VA Medical Center, New York, NY
238 Comparative Evaluation of the Performance of Abbott m2000rt Real-Time HIV-1 Assay for Measurement of
HIV-1 Plasma Viral Load
M. Vidya1, S. Saravanan1, Kartik K. Venkatesh2, P. Balakrishnan1, N. Kumarasamy1, K. G. Murugavel1, Sunil S.
Solomon1, Suniti Solomon1 and Kenneth H. Mayer2 ; 1YRG Centre for AIDS Research and Education, Voluntary
Health Services, Taramani, Chennai, India; and 2Brown University, RI, USA
239 Barriers to timely initiation of antiretroviral therapy among HIV infected children admitted to Mulago
Hospital Paediatric wards
Dr. Eleanor Namusoke, Joint Clinical Research Centre, Kampala, Uganda
240 Patient Retention in a University Hospital-based ART program in Uganda

Ouma Joseph
241 High Incidence Cohort of IDUs Infected with HIV with Low Genetics Diversity for HIV Vaccine Efficacy Trials
Sergey Verevochkin, St. Petersburg, Russia
242 Mycobacterium avium KatG protein (MAV_2753): a putative candidate for the serodiagnosis of MAC disease
Kapil Gupta1, Ajay Wanchu2, Romica Latawa1, S. Laal3, G.K. Khuller and Indu Verma1; 1Department of
Biochemistry; 2Department of Internal Medicine, Postgraduate Institute of Medical Education and Research,
Chandigarh, INDIA; and 3Department of Pathology, NYU Langone Medical Center, New York, USA
243 The Use of HIVQUAL as a Quality Improvement Tool for a Large Scale HIV/AIDS Public Health Program –
THE ACTION PROJECT, IHV Nigeria
U. Yakubu, U. Gebi, M. Babamaiyaki, I. Okoye, K. Falayajo, P. Dakum, J. Farley, W. Blattner, M. Etiebet, A. Zoakah
and M. Charurat; Institute of Human Virology, Abuja, Nigeria
244 Mannose Binding Lectin and its variants: susceptibility to HIV-1 and Schistosoma haematobium infection in
a rural Zimbabwean community
Rutendo B. L. Zinyama-Gutsire, National Institute of Health Research Ministry of Health and Child Welfare
Zimbabwe
245 Outcome of early infant diagnosis in exposed infants in Northern Nigeria

O. D. Adegoke, Z. Basir, J. Jumare, S. Sani, R. Enzama, S. Peters, A. Abimiku, W. A. Blattner; Institute of Human
Virology, Nigeria
246 Interface of public health implementation and research – Challenges and Prospects – IHVN/Action Project
Experience
O. Akinwande, Patrick Dakum, J. Farley, U. Gebi, C. Adebamowo, A. Abimiku, M. Charurat, W. Blattner; Institute
of Human Virology, Nigeria
247 The effect of chronic alcohol exposure in HIV/AIDS patients on antiretroviral drugs (Triomune30 –
AZT/3TC/NVP) in Uganda
Dr. Godfrey Sande Bbosa; Supervisors: Professor J. Ogwal-Okeng; Professor W. W. Anokbonggo and Associate
Professor B. D. Kyegombe; Department of Pharmacology and Therapeutics, Faculty of Medicine, Makerere
University, Kampala, Uganda
248 Telemedicine in Peru: training physicians responsible for the administration of highly active antiretroviral
therapy (HAART) in a developing country
Katiuska Castillo1, Raúl Gutiérrez1,3, Leslie Soto1, Alfonso Silva-Santisteban1, David Iglesias1, Alberto Guerra-
Garc´a1, Carlos Kiyan2, Carlos Seas1, Juan Echevarr´a1, Ciro Maguiña1 and Eduardo Gotuzzo1; 1Instituto de
Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima, Peru; 2Facultad de
Medicina, Universidad Peruana Cayetano Heredia, Lima, Peru; and 3Fogarty-NIH International Training and
Research Program in HIV/AIDS and TB, University of Miami, Miller School of Medicine
249 Efficacy of a Behavioral Intervention to Reduce HIV Risk among Female Sex Workers in Urumchi, China
Lin Han, Division of Policy and Information, NCAIDS, Beijing, China
250 Establishment of new Jurkat cell line Stably expressing HIV-1-Tat
Mohamed Ali Jarboui, William W. Hall and Virginie W. Gautier; Centre for Research in Infectious Diseases,
University College of Dublin, Dublin, Ireland
251 Migration, Pastoralists, HIV Infection and Access to Care: The Nomadic Fulani of Northern Nigeria
J. Jumare, A. G. Habib, U. Gebi, A. Zoakah, P. Dakum, J. Farley and W. A. Blattner; Institute of Human Virology,
Nigeria
252 Acceptance of and adherence to anti-retroviral therapy in Tanzania: the influence of lipodystrophy and of
traditional medicine
Sajida J. Kimambo
253 Changes of HIV risk behaviors of heroin drug users treated in methadone maintenance treatment clinics in
Guizhou province, China
Enwu Liu
254 Factors associated with development of Opportunistic Infections among patients on ART in Uganda
S. Muhumuza, J. Ouma, F. Semitala, E. Mbabazi and M. Kamya; Mulago-Mbarara Teaching Hospital’s Joint AIDS
Program (MJAP), Kampala, Uganda
255 HIV Related TB: Prospects and Challenges in a High Burden Area; A Case Study of Isoniazid Prophylaxis
(IPT) Utilization
B. M. Musa, U. Gebi, K. Falayajo, P. Dakum, J. Farley and W. Blattner; Institute of Human Virology, Nigeria
256 Dyslipidemia in HIV-1 infected individuals following generic efavirenz based HAART in a resource constrained setting
Dr. Sundaram Muthu, YRG CARE, Voluntary Health Services, Taramani, Chennai, India
257 Does toxicity to fixed dose stavudine, lamivudine and nevirapine regimen affect virologic suppression among
HIV infected adults at the Infectious Diseases Institute, Makerere University?
Fred C. Semitala1, Harriet Mayanja- Kizza1, Andrew Kambugu2, Agnes K. Kiragga2, Barbara Castelnuovo2, Robert
Kalyesubula1, Walter Schlech3, Robert Colebunders4, Keith McAdam2 and Moses R. Kamya1; 1Department of
Medicine Makerere University, Kampala, Uganda; 2Infectious Diseases Institute, Makerere University, Kampala,
Uganda; 3Dalhousie University Halifax, Canada; and 4Department of Clinical Sciences, Institute of Tropical
Medicine Antwerp, Belgium
258 Intra patient evolution of Human Immunodeficiency Virus Type 1 protease and reverse transcriptase in
antiretroviral naı̈ve patients
Uma Shanmugasundaram1, Kumarasamy Nagalingeswaran1, Kailapuri G. Murugavel1, Saravanan Shanmugam1,
Vidya Madhavan1, Sunil S. Solomon1, Suniti Solomon1, Kenneth H. Mayer2 and Balakrishnan Pachamuthu1;
1
Y. R. Gaitonde Centre for AIDS Research and Education, Chennai, India; and 2Department of Medicine, Brown
University/Miriam Hospital, Providence, RI, USA
259 Quality of life among end-of-life former commercial plasma donors infected with HIV and the care model in
rural Henan, China
Yu Sheng, Beijing P. R. China
260 Scaling Up PMTCT Access Using a Hub and Spoke Model in Nigeria, Sub-Sahara Africa
Dr. Akinmurele Timothy, Institute of Human Virology Nigeria
261 Impact of Community- and Home-Based Care on HIV/AIDS Patients and Their Families
Emily Hauwa Umaru, Institute of Human Virology, Nigeria
262 Prevalence of Norovirus infections in children from Iquitos, Peru

Daniel Velasquez Portocarrero, Jr. Enrique Barron, PERU
263 Adherence to highly active antiretroviral treatment and related factors in drug users with HIV/AIDS
Wang Honghong, School of Nursing, Central South University, Changsha, China
264 Immunologic recovery among HIV infected children on first line antiretroviral therapy in Kano, Nigeria
Bashir M. Zubayr
Author Index to Abstracts
A
Accolla 153 Dimitrov 184
Adegoke 245 Douek 122
Adeyemi 199 Douglas 115
Akinmurele 260
Akinwande 246 E
Akinyemi 217 Eldridge 121
Altaf 232 Elia 144
Alter 124 Engels 150
Amoroso 166 Eyzaguirre 189
Anastasopolou 104 Ezeaku 235
Attili 202
B F
Farthing 164
Badram 127
Fauci 146
Bagley 215
Felber 224
Bartlett 163
Finnegan 211
Bbosa 247
Berzofsky 175
Bewley 101 G
Biberfeld 181 Ganem 151
Bienasz 155 Gao 125
Brass 159 Garcia-Martinez 156
Bryant 223 Garzino-Demo 112
Buonaguro 183 Gebi 227
Buonaguro 210 Gershoni 161
Burkrinsky 195 Goldstein 126
Busari 233 Gougeon 114
Gray 180
C Greene 154
Cairo 208 Gu 116
Carbone 152 Guan 221
Castillo 248 Gupta 242
Chen 128
Choudhary 234 H
Cihlar 167 Han 249
Cocchi 187 Hanes 110
Contreras-Galindo 129 Hearl 132
Cummings 206 Heredia 198
Hokello 236
D Honghong 263
Dalgleish 174 Houghton 147
Daniels 130 Huang 222
de Noronha 157
Della Chiara 197 J
Denner 131 Jaffee 177
Denner 185 Jarboui 250
Denner 186 Jochmann 133
Derdeyn 107 Johnson 182
Desrosiers 145 Joseph 240
Desrosiers 226 Jumare 251
K R
Kelsoe 169 Reid 214
KewalRamani 117
Kijak 216 S
Kimambo 252 Sadowska 220
Klein 148 Sam 231
Klein 149 Schlom 176
Kurth 111 Selvam 200
Selvam 200a
L
Semitala 257
Lagenaur 105 Shankar 188
Latinovic 168 Shanmugasundaram 258
Lee 134 Shaw 108
Lehmann 205 Sheng 259
Leite 192 Siame 140
Leite 203 Subramaniam 100
Li 209 Subramanya 213
Li 135 Sundaram 204
Li, H. 237 Swanstrom 109
Lieberman 158 Synowski 194
Lisco 113
Liu 253 T
Lu 173
Luo 207 Temoshok 141
Tressler 165
M Tsibris 103
Mattapallil 136
Matucci 137 U
Melikian 102 Umaru 261
Michael 178
Moelling 118 V
Muhumuza 254 Vasiliver-Shamis 119
Mulenga 229 Veazey 106
Musa 255 Verevochkin 241
Muthu 256 Vidya 238
Visciano 212
N
Nabel 179
Nakimuli-Mpungu 228
W
Namusoke 239 Wainberg 162
Wald 193
Ndopu 218
Wald 193a
Newell 190
O Y
O’Brien 123 Yagamashi 120
Onigbogi 138 Yahaya 201
Onyango 139 Yahaya 219
Yakubu 243
P Yates 142
Pavlakis 170
Picker 172
Poli 160 Z
Poli 196 Zinyama-Gutsire 244
Pomerantz 225 Zipeto 143
Poonia 191 Zolla-Pazner 171
Portocarrero 262 Zubayr 264
100 3D structure of native HIV-1 gp120 trimers and
mechanisms of cellular entry
Sriram Subramaniam
Center for Cancer Research, NCI, NIH, Bethesda, MD 20892
We are using electron tomography and related methods in 3D electron microscopy to analyze the
structure of HIV-1 in the purified state, and at various stages of maturation in infected T-lymphocytes,
macrophages and mature dendritic cells. Using cryo-electron tomography combined with 3D image
classification and averaging, we have recently obtained the three-dimensional structures, at resolutions
of ;20 Å, of the trimeric envelope glycoprotein (Env) displayed on native HIV-1 in the unliganded state,
in complex with the broadly neutralizing antibody b12 and in a ternary complex with CD4 and the 17b
antibody. By fitting the known crystal structures of the monomeric gp120 core in the b12- and
CD4/17b-bound conformations into the density maps derived by electron tomography, we have
derived molecular models for the native HIV-1 gp120 trimer in unliganded and CD4-bound states. CD4
binding results in a major reorganization of the Env trimer, causing an outward rotation and
displacement of each gp120 monomer. This is coupled with a rearrangement of the gp41 region along
the central axis of the trimer, leading to closer contact between the viral and target cell membranes.
Our findings identify the structure and conformational changes of trimeric HIV-1 gp120 relevant to
antibody neutralization and attachment to target cells, and are likely to provide important clues to
vaccine design.
101 Chemokine Receptor CCR5 Mediates Resistance to
West Nile Virus Infection in Mouse and Man
Carole A. Bewley,1 Son N. Lam,1 Chih-Chin Huang,2 Priyamvada Acharya,2

Min Tang,2 Richard Wyatt,2 and Peter D. Kwong2
1
Laboratory of Bioorganic Chemistry, NIDDK, NIH; 2Vaccine Research Center, NIAID, NIH
The CCR5 co-receptor binds to CD4-activated HIV-1 gp120 surface envelope glycoprotein and
facilitates HIV-1 entry into cells. Key portions of CCR5 involved in gp120 binding include its N-terminus
and second extra cellular loop (ECL2). The N-terminus of CCR5 contains multiple tyrosine-sulfate (Tyr-
SO4) residues that are necessary for binding gp120, as do several antibodies that react with the co-
receptor-binding site on gp120. We have used a combination of NMR spectroscopy, X-ray
crystallography and molecular docking to solve the structure of the CCR5 N-terminus and the
tyrosine-sulfated antibody, 412d, in complex with gp120 and CD4. These analyses have revealed a highly
conserved Tyr-SO4-specific binding site on gp120 for which HIV-1 Entry inhibitors may be developed,
and provide the structural basis for post-translational mimicry between the immune system and an
HIV co-receptor. In ongoing studies we are using NMR and biochemical techniques to probe the
interactions between CCR5’s ECL-2 and gp120; results from these studies will also be discussed.
102 Visualization of HIV-1 entry pathways in cells
Gregory Melikian
Institute of Human Virology, 725 W. Lombard St., Baltimore, MD 21201
Viruses whose fusion proteins are activated by interactions with cellular receptors at neutral pH are
generally thought to fuse directly to a plasma membrane. However, recent evidence suggests that these
viruses may undergo receptor-mediated, pH-independent fusion with endosomes. To elucidate the HIV-1
entry pathways, we have developed an imaging assay capable of differentiating between surface fusion
(SF) and endosomal fusion (EF). Pseudoviruses bearing the pH-independent HIV-1 Env or the Avian
Sarcoma and Leukosis Virus (ASLV) Env, which has been shown to induce fusion in both receptor- and low
pH-dependent manner, were generated. Viral membrane and content redistribution during fusion was
visualized by co-labeled viruses with a core marker (Gag-GFP) and a red lipophilic dye (DiD). The Gag-GFP
cleavage by viral protease generated a smaller GFP-tagged fragment that was readily released from
virions upon their permeabilization. Double-labeled viruses were adhered to target cells expressing CD4
and coreceptors in the cold, and fusion was triggered by raising the temperature and monitored by a laser
scanning confocal microscopy. Fluorescent viruses undergoing SF are expected to loose their lipid marker
(hemifusion) and content marker (full fusion) due to dilution of these probes in the plasma membrane
and the cytosol, respectively. By contrast, full fusion with an endosome should be manifested in
disappearance of a content marker, but not a lipid dye. This strategy was validated by comparing the
outcomes of ASLV fusion in endosomes (the natural pathway) with fusion at the cell surface induced by
acidic pH. Imaging HIV-cell fusion (at neutral pH) detected both SF and EF events. Notably, HIV and ASLV
fusion with a plasma membrane was incomplete, resulting in lipid, but not viral content transfer within
the limited time frame of imaging experiments. In contrast, endosomal fusion events were associated
with the viral content release into the cytosol. These results indicate that HIV-1 can fuse both directly with
a plasma membrane and with endosomal compartments. It remains to be established which of these
alternative entry pathways is the one that leads to productive infection.
103 Tracking Dynamic HIV-1 Escape from CCR5
Antagonist Therapy in vivo by Deep Sequencing
Athe M. N. Tsibris
65 Landsdowne St, Room 435, Cambridge, MA 02139
High-throughput sequencing platforms provide an approach for detecting rare HIV-1 variants. We
applied this technology to the V3 loop-coding region of env in samples collected from 4 chronically
HIV-infected subjects who failed CCR5 antagonist (vicriviroc [VCV]) therapy. All subjects were receiving
VCV at the time of VF. Three time points were analyzed for each subject: study entry (wk 0), an
intermediate time point on study drug, and VF. HIV-1 RNA was extracted from plasma and subject-
specific primer sets were used to reverse transcribe and amplify plasma V3 loop-coding regions of env.
V3 amplicons were then submitted in a blinded fashion for deep sequencing and custom analysis.
Between 25,000–140,000 single genome sequences were obtained per subject per time point.
Profound baseline V3 loop sequence heterogeneity existed; predicted CXCR4-using populations
were identified in a largely CCR5-using population. V3 loop forms associated with subsequent
virologic failure, either through CXCR4 use or emergence of VCV resistance, were present as minor
variants at 0.8–2.8% of baseline samples. Deep sequencing provided a detailed view of both extreme
shifts in population frequencies toward these forms and the rapid evolutionary impact of VCV
selection. This degree of V3 loop sequence diversity has implications for vaccine design and the
optimal use of HIV-1 CCR5 antagonists.
104 A V3-independent pathway to resistance to small
molecule CCR5 inhibitors
Dr. Kleio Anastasopoulou

Weill Cornell Medical College, Dept. of Microbiology, 1300 York Ave, Rm: W-801, New York, NY 10065
CCR5-targeted entry inhibitors have now entered clinical practice for the treatment of HIV-1
infection. As with all antiretroviral drugs, the development of resistance should be anticipated and
understood. The genetic pathways to resistance to CCR5 inhibitors described to date have almost
invariably involved changes in the V3 region of the HIV-1 gp120 surface glycoprotein, irrespective of
the identity of the parental virus, of whether resistance developed in vivo or in vitro, and of
the method used to generate resistant variants. An exception is that no V3 changes were found in
a variant (D1/85.16) of the R5 subtype B primary isolate CC1/85 that was selected for resistance to
vicriviroc (VVC) during prolonged culture in PBMC (Marozsan et al. Virology 2005; 338: 182–199). The
resistant virus was found to be stable during 19 passages of culture in the absence of the drug, so there
are no obvious fitness costs associated with the development of VVC resistance. Moreover, the
phenotype of D1/85.16 is generally similar to those of other variants with critical resistance sequence
changes in V3. Thus the resistant variant has adapted to use the inhibitor-CCR5 complex, as well as the
free co-receptor. To locate the non-V3 resistance mutations, a VVC-resistant clone was generated from
the D1/85.16 isolate, and then used to produce chimeric viruses and site-directed mutants for further
analysis. The critical determinants of VVC resistance have now been shown to lie within a specific
region of the gp41 transmembrane glycoprotein. We are now investigating whether these amino acid
changes are necessary and sufficient for complete resistance, as well as trying to define how they
function. However, it is clear that although V3 changes are the dominant genetic pathway to CCR5
inhibitor resistance, there is a V3- (and gp120-) independent pathway that can create a similar
phenotype. Understanding how this pathway operates may assist in further defining structure-
function relationships within the trimeric HIV-1 Env complex.
105 sCD4-17b, an Engineered Bifunctional HIV-1
Neutralizing Protein: Potent Cross Clade Activity and
Potential Microbicide Use
Laurel A. Lagenaur,1,2 Vadim A. Villarroel,1 and Edward A. Berger1

1
NIAID, NIH, Bethesda, MD 20892, 2Osel, Inc., 4008 Burton Dr., Santa Clara, CA 95054, USA
We previously described sCD4-17b, an engineered single chain bifunctional protein that neutralizes
HIV-1 infection (Dey et al., J. Virol., 2003). sCD4-17b contains the first two domains of CD4 attached by
a flexible linker to a 17b mAb scFv, whose epitope overlaps the CD4i bridging sheet involved in
coreceptor binding. Based on the crystal structure of a gp120 core complexed to two-domain sCD4 and
the 17b Fab (Kwong et al., Nature, 1998), we originally anticipated that a 35 aa linker length would
readily span the 56 Å distance required for simultaneous binding of the sCD4 and 17b moieties, thus
neutralizing free virions. Linkers too short to enable simultaneous binding were predicted to be much
less active, perhaps equivalent to a mixture of unlinked sCD4 and 17b scFv.
We expressed sCD4-17b constructs containing linkers of various lengths, as well as free sCD4 and 17b
scFv. The purified proteins were tested against HIV-1 pseudotypes containing genetically diverse Envs in
the TZM-bl single-round neutralization assay. sCD4-17b constructs with linkers of 35 or 40 aa displayed
equivalently potent neutralization activities; as expected, a construct with a linker of only 5 aa was
significantly less potent, although it was more effective than equimolar concentrations of unlinked sCD4
plus 17b (Perhaps reflecting cross-binding of the sCD4 and 17b moieties to different gp120 subunits,
between different spikes on either the same or different virions). Unlinked sCD4 plus 17b scFv had
minimal activities over the same concentration ranges.
sCD4-17b with the 40 aa linker was tested against pseudotypes with Envs from genetically diverse
HIV-1 isolates. Amongst Envs from standardized clade B and C panels, as well as several from clades D, E,
and F, neutralization was observed in every case; the potencies were consistently high (IC50 2 – 40 nm,
corresponding to 0.1 – 2 mg/mL). Amongst Envs from early infection subtype A isolates previously shown
to be resistant to most broadly neutralizing mAbs (Blish et al., AIDS 2007), nearly all were neutralized by
sCD4-17b albeit with higher IC50 values (40–200 nM, corresponding to 2–10 mg/mL).
Neutralization of ‘‘live’’ virus via spreading infection in PBMC’s is currently being examined.
The combined breadth and potency of sCD4-17b exceeds that reported for any anti-HIV mAbs.
These findings suggest its potential use as a microbicide to prevent sexual transmission.
106 Early events in vaginal HIV transmission
Ronald S. Veazey1 and Thomas Hope2

1
Tulane National Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433, 2Northwestern
University, Chicago IL
Worldwide, the vast majority of HIV-1 cases occur through heterosexual transmission. Although the
initial events involved in vaginal transmission are uncertain, studies suggest that dendritic cells in the
vaginal epithelium may be involved in trapping viral particles on the surface, and transporting them to
susceptible memory CD4+ T lymphocytes in the mucosa. However, studies show that the vaginal
epithelium do not express known receptors for HIV attachment including CD4, CCR5, or DC-SIGN.
Although dendritic cells expressing CD1a are present in the epithelium, and CD4+CCR5+ memory cells
are abundant in the underlying lamina propria, the mechanisms involved in transport of HIV across the
epithelium to the underlying target cells remains debated. Using a photoactivateable HIV, we are
currently examining the earliest events in vaginal HIV transmission in rhesus macaques in vivo, and in
human vaginal/cervical explants ex vivo. These studies have shown that HIV passively penetrates
through layers of the squamous epithelium of the vagina, and thus gains access to intraepithelial
antigen presenting cells. Breaks or thinning of the epithelium increase the level of penetration, and
cervical mucus provides a protective barrier that traps most particles in the lumen. Although virus is
frequently observed to penetrate the vaginal epithelium, in experimental macaques, virus is rarely
found in the endocervix or upper reproductive tract. Combined, these data suggest that the vaginal
mucosa is the major site of HIV entry, and is likely the major site of viral transmission in the female
reproductive tract.
107 Escape from Neutralizing Antibody in Early Subtype
C HIV-1 Infection
Rong Rong,1 Bing Li,1 Rebecca M. Lynch,1 Joseph Mulenga,2 Susan Allen,1
Eric Hunter,1 Jerry L. Blackwell,1 and Cynthia A. Derdeyn1
1
Emory University, Atlanta, GA, USA; 2Zambia Blood Transfusion Service, Lusaka, Zambia
HIV-1 subtype C viruses circulate predominantly in India and sub-Saharan Africa, accounting for
most global infections. We previously demonstrated that 9 out of 11 subtype C infected
seroconvertors followed in a Zambian cohort rapidly developed high titer autologous neutralizing
antibody (Nab) against the infecting virus. Here we characterized the Nab sensitivity of individual
envelope (Env) glycoproteins that evolved during early infection in four of these seroconvertors.
Env genes were PCR amplified from longitudinal patient PBMC DNA and plasma samples using
single genome analysis, cloned into an expression vector, and screened for biological function using
a pseudovirus assay. Neutralization sensitivity of each Env pseudovirus was evaluated against
autologous plasma from longitudinal time points to identify variants that had escaped from
contemporaneous Nab. The molecular mechanism of Nab escape was then investigated in detail in
two subjects by replacing Env domains of the initial, Nab sensitive virus with those derived from 6 to
10 escape variants from the same patient.
The monophyletic founder virus in the newly infected subjects was rapidly replaced by
a succession of variants that were resistant to contemporaneous Nab but were subsequently
neutralized by a de novo antibody response. In one patient, the virus used multiple escape
mechanisms, even at a single time point. One of these escape pathways required sequence changes in
both gp120 and the gp41 ectodomain, while another involved only the V3-V5 region of gp120. The V1V2
domain, however, did not contribute to neutralization escape in this subject. In contrast, in the second
patient, the V1V2 domain played a major role in resistance in every Env analyzed. The changes in V1V2
conferred early and complete resistance against a monoclonal antibody isolated from the PBMC of this
patient four years later, but changes outside of V1V2 were required to escape from the polyclonal
antibody response.
Our data demonstrate that subtype C viruses use multiple mechanisms to escape from potent
autologous Nab, even within a single patient. These findings highlight the need to define and
understand potential escape mechanisms in strategies to induce protective Nab.
108 Identification of Full-length Transmitted HIV-1
Genomes Reveals Extraordinary Dynamics and
Precise Molecular Pathways of Early Virus
Diversification, Adaptation and Immune Evasion
George M. Shaw, MD, PhD

University of Alabama at Birmingham, 816 KAUL Bldg, 720 20th St. South, Birmingham, AL 35294-0024
Background: Identification of complete (9kb) genomes of HIV-1 viruses responsible for

productive clinical infection could be instrumental in elucidating viral properties and biological
events responsible for virus transmission and in characterizating subsequent virus evolution across the
proteome.
Methods: Recently, we developed a mathematical model of viral sequence evolution in acute HIV-
1 infection and an empirical dataset of 3449 complete HIV-1 subtype B env sequences derived by
single genome amplication (SGA)-direct amplicon sequencing that allowed us to infer the exact
nucleotide sequences of full-length env genes of transmitted and early founder viruses in 98 of 102
consecutively studied patients (Keele et al., PNAS 2008). Here, we applied the same experimental
strategy to the identification of full-length HIV-1 genomes in 12 subjects with acute or early subtype B
or C infection and characterized their early evolution.
Results: Prior to antibody seroconversion, viruses generally exhibited a Poisson distribution of
mutations and star-like phylogeny, which coalesced to inferred transmitted/early founder consensus
sequences at or near the estimated time of virus transmission. 11 of 12 subjects were productively
infected by a single virus (or virally-infected cell) and one was infected by two viruses. In four subjects,
we performed sequential SGA analyses of complete viral genomes over the first year of infection and
mapped each autologous CTL epitope recognized and escaped. We also mapped neutralizing
antibody (Nab) recognition and escape in Env. Immune recognition and escape occurred substantially
earlier for CTLs (;4 wks post-infection) than for Nabs (;12 wks post-infection). Within a two week
period beginning at peak viremia, virtually the entire body’s population of productively infected cells
was replaced by cells infected with viruses containing CTL escape mutations that were confirmed
phenotypically. Complete replacement of transmitted wildtype virus by Nab escape virus occurred
between 8 and 12 weeks post-infection.
Conclusions: Identification of transmitted/founder full-length genomes allows for a comprehen-
sive and dynamic assessment of immune recognition and escape in HIV-1 infected humans, including
individuals immunized with candidate HIV-1 vaccines who experience breakthrough infections. We are
currently conducting similar studies in monkeys infected intrarectally by SIVmac251 or SIVsmE660 to
better define this animal model system and its relationship to humans infected mucosally by HIV-1.
109 Complexity of the Transmitted Virus and
Compartmentalization In Its Seminal Source
Ronald Swanstrom
UNC Center For AIDS Research
The transmission event has become a focal point for understanding the earliest steps of infection
with HIV-1. Most transmission events occur during unprotected sex. Thus there is special interest in the
virus that crosses the mucosal surface and also the source of virus in genital secretions, as these two
sites define the bottleneck that occurs during transmission. We have employed the single genome
amplification strategy to examine the env gene in the transmitted virus and also in the blood and
semen of chronically infected men, all in the context of subtype C infection. A comparison of
sequences from transmitted viruses to those found in chronic infection suggests differences in the
proteins encoded by these env genes. While most infections are initiated with the transmission of
a single variant, approximately 20% of infections are the result of infection with two or more variants.
Multiple variants were found in both men and women, and the frequency of the presence of multiple
variants is inconsistent with these representing independent transmission events of low probability.
Thus transmission rates must be transiently high or the transmission of multiple variants must
represent linked events. Comparison of virus in blood plasma and seminal plasma of chronically
infected men shows frequent clonal amplification of virus in the seminal tract, changing the
composition of the viral population from that seen in the blood. In some cases there is more extensive
compartmentalization, indicative of sustained independent replication in the seminal tract. These
results show the presence of genetic differences in both the donor and the recipient that affect the
nature of the transmitted virus. This work represents a large collaborative effort within the CHAVI.
110 HIV permeation efficiency in human cervicovaginal
mucus
Justin Hanes
Johns Hopkins University, 3400 N. Charles St., MD 221, Baltimore, MD 21218
HIV is transmitted sexually with unusually low efficiency (typically ,1 per ;1000 coital acts). To
reliably infect a primate model for HIV, ;10,000-fold more virus must be delivered vaginally than
intravenously. However, the vaginal mechanisms that help protect against HIV are poorly understood.
Here we report that human cervicovaginal mucus (CVM), obtained from donors with normal
lactobacillus-dominated vaginal flora, efficiently traps HIV, causing it to diffuse more than 1000-fold
slower than it does in water. Lactobacilli acidify CVM to pH ;4 by continuously producing lactic acid.
At this acidic pH we find that lactic acid, but not HCl, abolishes the negative surface charge on HIV. In
contrast, in CVM neutralized to pH 6–7, as occurs when semen temporarily neutralizes the vagina, HIV
maintains its native surface charge and diffuses rapidly, only 15-fold slower than in water. Thus,
methods that can maintain the native CVM acidity during coitus may contribute to both vaginal and
penile protection by trapping HIV before it can reach target cells. Our results reveal that CVM likely
plays an important but currently unappreciated role in decreasing the rate of HIV sexual transmission.
111 Reconstitution and Characterization of a Human
Endogenous Retrovirus with the Original Sequence
at the Time of Integration
Reinhard Kurth, Nadine Beimforde, Kirsten Hanke, and Norbert Bannert

Robert Koch-Institute, Nordufer 20, D-13353 Berlin, Germany
Human endogenous retroviruses (HERVs) are remnants of infectious exogenous retroviruses that
invaded the germ line of our ancestors. These elements have since been transmitted vertically from
generation to generation acquiring postinsertional mutations. In contrast to several other species, no
infectious endogenous retroviruses have so far been identified in the human genome, although there
is evidence for integration of active and replication competent HERV-K(HML-2) proviruses in our very
recent evolutionary history and a still ongoing particle production in some tissues has been
documented. The presence of multiple proviruses in the chromosomes and the expression of
functional viral proteins seems to have various pathogenic and in some instances also beneficial
implications for the host. To facilitate molecular and functional studies of HERV elements, its
pathogenic potential and the human factors restricting their proliferation, we have cloned HERV-K113,
one of the youngest and most complete of the HERV-K(HML-2) proviruses. Subsequently, we identified
nonsynonymous postinsertional mutations in the HERV-K113 sequence by aligning it with other
human specific HERV-K elements. These randomly acquired mutations were reverted by site-directed
mutagenesis, reconstituting a provirus encoding proteins which the virus produced at the time of its
integration. The properties of the viral proteins and of the generated viral particles as well as several
aspects of its lifecycle will be discussed in the light of recent findings obtained by others using HERV-K
consensus sequences.
112 hBD2 inhibits HIV via CCR6, a receptor expressed on
memory, Dendritic and Th17 cells
Mark K. Lafferty, Lingling Sun, Wuyuan Lu, and Alfredo

Garzino-Demo
Institute of Human Virology and Dept. of Microbiology and Immunology, University of Maryland
School of Medicine, Baltimore MD 21201
Human beta defensins (hBD) are antimicrobial peptides that are produced by epithelial cells, and
are components of innate immunity in mucosae. hBD2 and 3 inhibit HIV infection. Based on the
properties of these cationic peptides, which inhibit infectious agents by disrupting their negatively-
charged membranes or to glycan moieties on their glycoproteins we expected that hBD2 would exert
its HIV-suppressive activity by inactivating HIV, binding to its envelope. While we observed some
degree of virus inactivation, we found that hBD2 inhibits HIV also after virus entry. Since hBD2 binds to,
and triggers signaling via the chemokine receptor CCR6, we tested the hypothesis that CCR6 mediates
the antiviral activity of hBD2. Three lines of evidence support this hypothesis. First, MIP-3a (CCL20), the
chemokine ligand for CCR6, induces antiviral activity. Second, hBD2 loses efficacy when tested in
CCR6- cells, both in single cycle and spreading infectivity assays. Third, Bordetella Pertussis toxin, an
inhibitor of chemokine receptor –mediated intracellular signaling, abrogates the antiviral activity of
hBD2 and MIP-3a. Our mechanistic studies show that HIV inhibition occurs at an early stage, prior to
completion of reverse transcription. In summary, this is a novel antiviral activity mediated by
a chemokine receptor.
CCR6 is expressed in cells that are highly relevant to HIV infection, mucosally and in the periphery:
memory CCR5+ T lymphocytes, dendritic cells and, based on our data, on LPS-activated macrophages.
Interestingly, CCR6 is one of the markers for the Th17 subset of T lymphocytes. Published reports have
shown that CCR6+ and Th17 cells are specifically lost in the course of progressive HIV disease or in
primate models of AIDS. Thus, a mechanism that protects CCR6 cells from infection may yield novel
preventive or therapeutic approaches to HIV infection.
113 The anti-herpetic drug acyclovir suppresses HIV-1 in
herpesvirus-infected human tissues after
conversion into a nucleoside reverse transcriptase
inhibitor
Andrea Lisco
Section of Intercellular Interactions, Eunice Kennedy Shriver National Institute of Child Health and
Human Development, 10 Center Drive, Bldg.10, Room 9D58, 20892, Bethesda, MD
Background: Ever-new, safe, efficient and inexpensive strategies against HIV are needed because
of the increase of HIV-1 epidemics and the emergence of drug-resistant viruses. Such strategies have
been developed against HSV, which can be efficiently treated with acyclovir (ACV). The anti-herpetic
specificity of ACV is primarily based on the unique ability of HHV-encoded kinases to phosphorylate
ACV to its monophosphate derivative, which is subsequently converted into the antivirally active
ACVtriphosphate (TP). Consistently with its highly restricted anti-herpetic activity, ACV is not used as
a direct HIV-1 inhibitor.
Methods: We measured the effect of ACV-TP on HIV-1 reverse transcriptase (RT) activity in a cell-
free system using a gel-based assay. Various HIV-1-infected human tissues were treated ex vivo with
ACV and newly synthesized ACV-based prodrugs containing a masked phosphate group. We
monitored viral replication in tissues and cell lines using real-time PCR and ELISA.
Results: We demonstrate the direct inhibitory effect of ACV-TP, but not of ACV itself, on isolated
HIV-1 RT. The kinetics of incorporation of ACV-TP by HIV-1 RT, evidence of its phosphorolytic excision,
and the kinetics of formation and dissolution of a dead–end-complex with HIV-1 RT demonstrate that
ACV-TP acts as a nucleoside RT inhibitor. Consequently, ACV suppresses HIV-1 replication in various
human tissues and in T-cell line cultures carrying various HHVs that are capable of phosphorylating
ACV. Furthermore, we demonstrate that monophosphorylated ACV-based prodrugs, which bypass the
requirement for HHV-mediated phosphorylation, are able to inhibit HIV-1 replication irrespective of
the presence of HHVs.
Conclusions: In human tissues coinfected with various herpesviruses, ACV is activated into an HIV
RT inhibitor that suppresses HIV-1. This newly discovered mechanism of HIV suppression by ACV may
help to explain the results of several recent ACV trials in HSV-2/HIV-1-coinfected patients. The
discovery of the antiretroviral activity of ACV-monophosphate prodrugs can promote the
development of a new class of HIV-1 RT inhibitors.
114 NK cells contribute to the constitution of HIV
reservoirs in dendritic cells
Dr. Marie-Lise GOUGEON

Institut Pasteur, 28 rue du Dr. Roux, Paris 75015, France
Early stages of viral infections are associated with local recruitment and activation of effectors of
innate immunity, i.e. NK cells and DCs. DCs are essential for both antigen-presentation and activation
of na¨ve CD4+ T cells, and further Th1 polarization. DCs also constitute early targets for HIV. NK-DC
crosstalk is important for DC homeostasis and maturation, but its contribution to susceptibility of DCs
to infection, and to DC-dependent Th1 polarisation is unknown. We addressed this question in the
present study.
We report that HIV-infection of immature DC did not induce their maturation, as evaluated by the
coexpression of CD86 and HLA-DR, while their coculture with activated NK cells did. NK-induced
maturation of DC was inhibited by polyclonal antibodies specific for HMGB1 molecule or by
glycyrrhizin, suggesting an important role of the pro-inflammatory cytokine HMGB1 in this process.
Interestingly, activated NK were also able to polarize the production of IL-12 by DC that in turn induced
a T helper
1-like immune response when cocultured with naive CD4+ T cells. In contrast, infected DCs were no
more susceptible to NK-dependent IL-12 polarization, and thus no more able to induce a Th1 response.
In addition, NK-dependent DC maturation was associated with an increased production of HIV-1 p24
and an increased expression of proviral DNA by DC.
Altogether, our results show that activated NK cells 1- induce the maturation of DC through an
HMGB-1-dependent mechanism; 2- trigger IL-12 production by DC and further Th1 polarization;
3- increase HIV production and the constitution of viral reservoirs in infected DC; 4- facilitate HIV-1
transmission from DC to T cells.
115 HIV Vpu complexes with bTrCP to direct the
degradation of the virus release inhibitor BST-2
(Tetherin)
Janet L. Douglas, Kasinath Viswanathan, Matthew N. McCarroll, Jean K. Gustin,

Klaus Fruh, and Ashlee V. Moses
Oregon Health & Science University-VGTI, 505 NW 185th Ave, Beaverton, OR 97006
The primary roles attributed to the HIV-1 Vpu protein are the targeted downregulation of the viral
receptor CD4 and the enhancement of virion release. With regards to CD4 degradation, Vpu has been
shown to act as an adaptor linking CD4 with the ubiquitin/proteasome machinery via interaction with
the F-box protein bTrCP. This interaction was previously thought to be dispensable for Vpu’s virion
release function. In an attempt to identify other cellular bTrCP-dependent Vpu targets, we performed
quantitative proteomics on the plasma membrane fraction of Hela cells expressing either a wildtype
Vpu, or a Vpu double point mutant (S52N/S56N) that no longer binds to bTrCP. We identified one
cellular protein, BST-2 (CD317), that was significantly downregulated by wildtype Vpu, but not the
mutant. Recently, BST-2 has been identified as the IFN-inducible cellular factor Tetherin, which limits
HIV virion release in the absence of Vpu. We address here the mechanism of Vpu-mediated BST-2
downregulation. To verify the biological relevance of this phenotype, we show that in T-cells infected
with HIV-1, BST-2 downregulation does indeed occur and is Vpu dependent. Interestingly, HIV-2, which
does not encode Vpu, did not downregulate BST-2. Because our screen was designed to identify those
proteins downregulated by Vpu in a bTrCP-dependent manner, we next sought to determine if Vpu
targets BST-2 for degradation. We show via immunofluorescence and co-immunoprecipitations that
both wildtype and mutant Vpu are able to interact with BST-2, but only wildtype Vpu results in BST-2
degradation. Further support for bTrCP’s role was provided by showing that the dominant negative
mutant DF-bTrCP, which can no longer interact with the SCF ubiquitin ligase complex, blocks Vpu’s
ability to downregulate BST-2. In addition, treatment of cells with either the proteasome inhibitor
MG132 or the lysosome acidification inhibitor concanamycin A reduces BST-2 downregulation by
wildtype Vpu. Taken together, these data support the hypothesis that, similar to its role in CD4
degradation, Vpu acts as an adaptor molecule linking BST-2 to the cellular ubiquitination machinery
via bTrCP. This interaction then leads to the degradation of BST-2, which prevents viral tethering and
therefore results in enhanced virion release.
116 Molecular studies of HIV-1 Rev function
L. Gu, V. W. Gautier, N. Sheehy, T. Tsuji, H. Hayakawa, and W. W. Hall

Centre for Research in Infectious Diseases (CRID), University College Dublin, Dublin 4, Ireland
The HIV-1 regulatory protein Rev is critical for viral replication and modulates nuclear export of
partially spliced and unspliced viral transcripts. Its function requires Rev to be actively imported and
exported from the nucleus via nuclear pore complexes, a process mediated by recognition of its
Nuclear Localisation Signal (NLS) domain by importin-b and its Nuclear Export Signal domain by CRM-1.
Here we report that the human I-mfa domain-containing protein, HIC, modulates Rev nuclear
localisation and function by physically interacting with its NLS domain, impairing nuclear import by
rendering the NLS inaccessible to importin-b.
Co-immunoprecipitation studies and GST- Pulldown assays demonstrated that HIV-1 Rev
physically interacts with HIC via its NLS domain. Ectopic expression of HIC resulted in the cytoplasmic
redistribution of Rev with a concomitant reduction in its nuclear accumulation. This correlated with
a decrease in Rev activity as shown by the reporter gene assay. In vivo co-localisation studies in the
presence of Leptomycin B suggested that HIC inhibits Rev nuclear import instead of promoting its
nuclear export. This was subsequently confirmed by in vitro nuclear import assays, where competitive
amount of recombinant 63His-HIC specifically inhibited Rev nuclear import by Importin-b.
Functionally, the cytoplasmic sequestration of Rev may represent a novel mechanism for the
control of Rev function and ultimately the regulation of HIV-1 replication, and may provide a new
target for therapeutic intervention.
117 CA-Dependent HIV-1 Nuclear Import Relies on
Transportin-SR2
Kyeongeun Lee, Thomas D. Martin, Zandrea Ambrose, Alok Mulky, and

Vineet N. KewalRamani
HIV Drug Resistance Program, National Cancer Institute, Frederick, MD 21702
We have previously shown that C-terminally truncated forms of cleavage and polyadenylation
factor 6 (CPSF6), an SR family protein, interfere with infection by HIV-1 and SIV, but not MLV. HIV-1
entry and reverse transcription are not impaired in the presence of antiviral CPSF6, but nuclear forms
of the vDNA are diminished. Strikingly, growth arrest of cells expressing antiviral CPSF6 intensifies the
restriction of HIV-1 to over two orders of magnitude. A single amino acid change in HIV-1 CA, N74D,
overcomes the restriction by antiviral CPSF6. While N74D HIV-1 can efficiently infect transformed cell
lines and primary T cells in the presence or absence of antiviral CPSF6, this mutant virus is blocked at an
early stage in the infection of macrophages. These data suggested that antiviral CPSF6 interfered with
a nuclear entry pathway relevant to HIV-1 infection of primary target cells. To better understand how
antiviral CPSF6 prevents HIV-1 nuclear entry, we examined whether a recently described HIV-1
dependency factor, TNPO3, identified in a genome-wide siRNA screen by Elledge and colleagues was
relevant to infection by wild-type (WT) but not N74D HIV-1. Whereas siRNA knockdown of endogenous
CPSF6 or its binding factor CPSF5 did not impair HIV-1 infection, knockdown of the transportin-SR2
spliced isoform of TNPO3 specifically restricted WT but not N74D HIV-1. Notably, growth-arrest of cells
knocked down for transportin-SR2 intensified the block to WT HIV-1 infection, phenocopying the
restriction by antiviral CSPF6. These data reinforce a role for transportin-SR2, a non-‘‘NLS’’ karyopherin, in
the early replication of HIV-1 and suggest that its function may be particularly relevant for HIV-1
infection of nondividing cells. Our findings provide the first genetic evidence that HIV-1 CA regulates
interactions with nuclear pore associated transport factors.
118 Self-inactivation of HIV by its own RT/RNase H
K. Moelling, A. Matskevich, L. Wittmer-Elzaouk, J. Heinrich, J.-S. Jung,T. Kwok,

and S. Mathur
Section Viruses and Cancer, University of Zurich, Switzerland
We have designed a method to inactivate HIV infectivity in the extracellular environment by

activating the virion-associated Ribonuclease H. This enzyme has not been targeted for antiretroviral
therapy. The effect is mediated by a short hairpin-loop DNA targeted to the highly conserved
polypurine tract, the PPT (1–5). Treatment of HIV-infected serum isolated from patients and African
strains showed about 100fold reduction of infectivity in 33% of the cases. A recombinant retrovirus
was applied to the mouse vagina and showed a statistically significant reduction of viral RNA by
prophylactic or therapeutic regimens of DNA application. Previously we showed in a retroviral mouse
model that the viral load in the blood was reduced, disease progression delayed and infection prevented
(6). We compared the effects of the hairpin-loop-structured DNA to antisense DNA and siRNA and find
clear differences. The mechanism of uptake is independent of TLR9 and not yet understood.
(1, 2) Jendis et al., AIDS Res. Hum. Retroviruses (1996, 98); (3) Moelling et al., FEBS Letters (2006); (4) Matskevich et al., AIDS Res. Hum. Retroviruses (2006);
(5) Moelling et al. CSH Symp. 71 (2006); (6) Matzen et al. Nature Biotech. 23, 663 (2007).
119 HIV-1 envelope gp120 induces a stop signal and
virological synapse formation in non-infected CD4+
T cells
Gaia Vasiliver-Shamis, 1 Michael Cho, 2 Michael L. Dustin, 1 and

Catarina E. Hioe3
1
Skirball Institute for Biomolecular Medicine, New York University School of Medicine; 2Case Western
Reserve University School of Medicine; 3VA New York Harbor Health Care System and New York
University School of Medicine
HIV-1-infected T cells form a virological synapse (VS) with non-infected CD4+ T cells in order to
efficiently transfer HIV-1 virions from cell to cell. The virological synapse is a specialized cellular
junction that is similar in some respects to the immunological synapse involved in T cell activation and
effector functions mediated by the T cell antigen receptor. The immunological synapse stops T cell
migration to allow sustained interaction between T cell and antigen-presenting cells. In this study we
have asked whether HIV-1 envelope gp120 presented on a surface to mimic an HIV-1 infected T cell
also delivers a stop signal and if this is sufficient to induce a virological synapse. Using transmigration
assay we demonstrate that HIV-1 gp120-presenting surfaces arrested the migration of primary
activated CD4+ T cells that occurs spontaneously in the presence of ICAM-1. The VS dynamics were
evaluated using real-time fluorescence imaging of non-infected CD4+ T cells on a glass-supported
planar bilayer containing HIV-1 gp120 and ICAM-1. The data reveal for the first time high-resolution en-
face images of VS that is characterized by segregated supramolecular structures with a central cluster
of HIV-1 gp120 surrounded by a ring of ICAM-1. Interestingly, unlike the immunological synapse
that can be stable for .1 hour, the VS was formed transiently with initiation of T cell migration within
30 minutes. Both the stop signal and VS formation required HIV-1 gp120 interaction with CD4 but not
the chemokine receptor. Thus, HIV-1 gp120-presenting surfaces induce a transient stop signal and
supramolecular segregation in non-infected CD4+ T cells by a CD4-dependent manner. The HIV-1
gp120-induced transient arrest and VS assembly may constitute an efficient mechanism for virus
transfer from infected cells to target cells without incurring cell-cell fusion that terminates the virus life
cycle.
120 Promoter-targeted shRNA driven by retroviral
vector achieves long-term repression of HIV-1
replication
Makoto Yamagishi, Takaomi Ishida, Ariko Miyake, Kazuo Suzuki, and

Toshiki Watanabe
Laboratory of Tumor Cell Biology, Department of Medical Genome Sciences, Graduate School of
Frontier Sciences, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-
ku, Tokyo 108-8639, Japan
Regulation of gene expression by siRNA is a new therapeutic approach to a variety of viruses and
related diseases. In this conference, we will report an improved procedure using an shRNA targeted to
HIV-1 promoter region, which induces transcriptional gene silencing (TGS). Recently, some studies
demonstrated that promoter-targeted siRNA induces TGS in mammalian cells. However, the
mechanism of TGS remains unclear. When we use siRNA as a therapeutic tool against viruses, for
example HIV-1, siRNA-mediated mRNA degradation can transiently suppress HIV-1. However, rapid
emergence of ÔsiRNA escape mutantsÕ has been observed in vitro, since HIV-1 can quickly adapt to
environmental pressures. In this study, we investigated the efficiency of inhibition and the
mechanisms of TGS induced by shRNA-directed TGS of HIV-1 provirus integrated in Tcell. For this
purpose, we prepared shRNAs homologous to HIV-1 U3 region and constructed retroviral expression
vectors. Using these vectors, we established shRNA-expressing CD4 (+) T-cell lines. We measured the
levels of HIV-1 replication and found that an shRNA targeting NF-kB binding site within LTR inhibited
HIV-1 gene expression and its replication for unexpectedly long periods (more than 3 months) without
escape mutants. Nuclear run-on assays revealed direct inhibition of transcription of HIV-1 genes, indicating
that TGS was induced by this shRNA. Moreover, the shRNA inhibited reactivation of HIV-1 in latently
infected cell lines. To elucidate the mechanism of TGS against HIV-1 promoter, we studied reactivation of
HIV-1 replication by RT-PCR in cells where HIV-1 replication is suppressed by the shRNA, using HDAC
inhibitor, TSA, or DNA demethylation agent, 5-AzaC. Furthermore, chromatin immunoprecipitation assays
demonstrated induction of H3K27 trimetylation and subsequent facultative heterochromatin formation of
shRNA-targeted promoter. These data suggested that shRNA targeted to HIV-1 promoter region induced
TGS in an epigenetics-dependent manner, resulting in longterm suppression of HIV-1 replication.
121 Plasmid DNA- and live viral vector-based vaccine
approaches for the treatment and prevention of
HIV infection
John H. Eldridge, PhD

Profectus Biosciences, Inc.
Almost two decades ago, it was first demonstrated that the direct injection of a plasmid DNA
vaccine encoding a foreign antigen resulted in plasmid uptake, protein expression, and the induction
of an antigen-specific cellular and humoral immune response. In the interim, the ability of DNA
vaccine-elicited immune responses to protect against viral and bacterial infections, parasites, cancers,
and autoimmune diseases has been well-documented in numerous animal models. Phase I human
clinical trials have shown that experimental DNA vaccines are safe and well tolerated, however, these
preliminary studies indicate that measures must be taken to improve vaccine immunogenicity. One
particularly promising approach to improve the immunogenicity of DNA vaccines is through the co-
delivery of cytokine expression plasmids as genetic adjuvants. Studies in a variety of animal models
clearly demonstrate that plasmid DNA encoded immunomodulatory cytokines (IL-12, IL-15) not only
alter the magnitude and direction of the DNA vaccine-elicited immune response, but can also improve
vaccine efficacy. Another approach to improve the immunogenicity of DNA vaccines is through the
optimization of in vivo pDNA delivery. Recently, in vivo electroporation, i.e. the application of short
electrical pulses, has been shown to enhance gene delivery and dramatically improve the induction of
vaccine antigen-specific cell-mediated and humoral immune responses.
Recombinant vesicular stomatitis virus (rVSV) vectors expressing HIV/SIV antigens elicit robust immune
responses in rhesus macaques, significantly reduced viral load and alter disease pathogenesis following
challenge with the simian-human immunodeficiency virus SHIV89.6P. However, when tested in a stringent
non-human primate neurovirulence model, the lab-adapted rVSV vector appeared to be inadequately atten-
uated for use in humans. To address this concern, a variety of strategies were employed to produce more
attenuated vectors, and these further attenuated vectors displayed markedly reduced neurovirulence in
murine, ferret and non-human primate models, while retaining levels of immunogenicity equivalent to
the more virulent rVSV lab-adapted vector. As a result of these efforts, a further attenuated replication
competent rVSV clinical trial candidate with an appropriate safety profile that retains the ability to
propagate to high levels in vitro and elicit robust immune responses in vivo has been identified.
Collectively, improved pDNA expression vectors, co-delivery of plasmid-based immunomodu-
lators and improved DNA delivery when used in combination with replication competent viral vectors
in heterologus prime-boost vaccination regimens have the potential to bring the field closer to the
design and development of an efficacious DNA vaccine for the prevention and/or treatment of HIV-1
infection.
122 Differential CD4 T cell subset depletion at mucosal
sites in HIV infection
Daniel C. Douek, MD, PhD

National Institutes of Health, Vaccine Research Center, 40 Convent Dr., MSC 3022, Bethesda, MD 20892
Acute HIV infection is characterized by massive loss of CD4 T cells from the gastrointestinal (GI)
tract. Th17 cells are critical in the defense against microbes, particularly at mucosal surfaces. We
analyzed Th17 cells in the blood, GI tract and broncheoalveolar lavage (BAL) of HIV-infected and
uninfected humans, and SIV-infected and uninfected sooty mangabeys. We found that (i) human Th17
cells are specific for extracellular bacterial and fungal antigens, but not common viral antigens; (ii) Th17
cells are infected by HIV in vivo, but not preferentially so; (iii) CD4 T cells in blood of HIV-infected
individuals are skewed away from a Th17 phenotype towards a Th1 phenotype with cellular maturation;
(iv) there is significant loss of Th17 cells in the GI tract of HIV-infected individuals (v) Th17 cells are not
preferentially lost from the BAL of HIV-infected individuals and (vi) SIV-infected sooty mangabeys
maintain healthy frequencies of Th17 cells in the blood and GI tract. These observations further elucidate
the immunodeficiency of HIV disease and may provide a mechanistic basis for the mucosal barrier
breakdown that characterizes HIV infection. Finally, these data may help account for the non-progressive
nature of non-pathogenic SIV infection in sooty mangabeys.
123 AIDS Restriction Genes-Genome Wide Association
Study
Stephen J. O’Brien
Laboratory of Genomic Diversity, NCI
The screening of common genetic polymorphisms among candidate genes for AIDS pathology in
HIV exposed cohort populations has led to the description of twenty AIDS Restriction Genes (ARGs),
variants that affect susceptibility to HIV infection or to AIDS progression. All of these ARGs validated by
our group have been discovered using the candidate gene approach. We have recently undertaken
a genome wide association analysis (GWAS) for some 6500 study participants registered
in longitudinal AIDS cohort populations using he Affymetrix 6.0 genotyping array which
screens .900,000 human SNPs and as many features for copy number variation ( CNV) . The genetic
associations were analyzed using both single monotypic AIDS association as well as multivariate
hypothesis analyses using non-independent genetic association tests as internal replication . My
presentation will demonstrate the gene discovery approaches and the results of a GWAS screen for
undiscovered ARGs that were revealed by this GWAS.
124 NK cells: friends or foes?
Galit Alter, 1 Daniel Kavanagh, 1 Suzannah Rihn, 1 Maureen Martin, 2

Mary Carrington,2 and Marcus Altfeld1
1
Partners AIDS Research Center and Infectious Disease Unit, Massachusetts General Hospital and
Division of AIDS, Harvard Medical School Boston, MA; 2Basic Research Program, SAIC-Frederick, Inc.,
Laboratory of Genomic Diversity, Frederick, MD
Decline of peak viremia during acute HIV-1 infection occurs prior to the development of vigorous
adaptive immunity and the level of decline correlates inversely with rate of AIDS progression,
implicating a potential role for the innate immune response in determining disease outcome. Natural
Killer (NK) cells are critical both in the early control of various viral infections but also play a critical role
in inducing a vigorous antiviral adaptive immune response. The combined expression of the activating
NK cell receptor KIR3DS1 and its presumed ligand HLA-B Bw4-80I has been associated with slow
progression to AIDS, however it is uncertain whether subjects with this genotype exhibit qualitatively
different patterns of NK cell expansions and contractions during acute infection that may contribute to
differences in patterning of the adaptive immune response. This presentation will highlight our recent
findings demonstrating that NK cells play a role in both promoting pathogenic events as well as
protecting against disease progression in the presence of protective KIR/HLA genotypes in HIV
infection.
125 Immunogenetic polymorphisms and their effects on
AIDS progression
Xiaojiang Gao
Cancer and Inflammation Program, SAIC-Frederick, National Cancer Institute, Frederick, MD, USA
For the past 10 years we have been investigating the differential effects of genetic diversity on
AIDS pathogenesis and HIV infection. In particular, we are interested in two highly polymorphic
immunogenetic systems, namely human leukocyte antigen (HLA) and killer immunoglobulin-like
receptor (KIR). The allelic polymorphism of HLA genes and chromosomal organization of KIR genes
were characterized in multiple AIDS cohorts to examine their effects on AIDS progression and HIV-1
infection. HLA zygosity and three HLA-B alleles show a consistent association with AIDS: HLA-B*35Px
shows a susceptible effect associated with a more rapid progression to AIDS, whereas HLA-B*27, B*57
and HLA heterozygosity show protective effects associated with a slower disease progression. Most
immunogenetic effects are detected at the stage of CD4 T-cell depletion, but the B27 protection is
most significant in the later stages of AIDS-defining illness. The same three HLA alleles are
also associated with different risks for HIV transmission: hemophiliac patients with B*35Px were more
likely to transmit HIV-1 to their female partners whereas patients with B*27 and B*57 were less likely to
transmit the virus. The protective HLA alleles also correlate with a lower level of HIV viral loads
compared to the susceptible alleles. Therefore, one mechanism to explain the observed HLA
associations with AIDS is their influence on viral load control. The newly described KIR system is an
important component of the innate immune mechanism. These polymorphic receptors regulate NK
killing of abnormal cells through recognition of class I HLA expression. Therefore KIR may be involved
in many of the diseases for which an HLA influence has been identified. Genetic epistasis between two
functionally-relevant immunogenetic systems was observed: the combination of KIR3DS1 and its
putative ligand HLA-Bw480I show a significant protection for AIDS progression, but either alone has
no effect. Our results demonstrate that genetic polymorphisms of both acquired and innate immune
systems influence AIDS pathogenesis and the disease progression.
This project has been funded in whole or in part with federal funds from the National Cancer
Institute, National Institutes of Health, under contract N01-CO-12400. The content of this publication
does not necessarily reflect the views or policies of the Department of Health and Human Services, nor
does mention of trade names, commercial products, or organizations imply endorsement by the
U.S. Government.
126 David Goldstein
Abstracts not available at time of printing
127 HTLV-I infection of WE17/10 CD4+ cell line leads to
progressive alteration of Ca2+ influx that eventually
results in loss of CD7 expression and activation of an
antiapoptotic pathway involving AKT and BAD
which paves the way for malignant transformation
H. Akl, B. Badran, G. Dobirta, N. E. Zein, F. Bex, C. Sotiriou, K. E. Willard-

Gallo, A. Burny, and P. Martiat
Laboratory of Experimental Hematology, Jules Bordet Institute, University of Brussels (ULB), Brussels,
Belgium
Adult T-cell leukemia/lymphoma (ATLL) is a malignancy slowly emerging from human T-cell
leukemia virus type 1 (HTLV-I)-infected mature CD45 T-cells. To characterize the molecular
modifications induced by HTLV-I infection, we compared HTLVI-infected WE17/10 cells with control
cells, using micro-arrays. Many calcium-related genes were progressively downmodulated over
a period of 2 years. Infected cells acquired a profound decrease of intracellular calcium levels in
response to ionomycin, timely correlated with decreased CD7 expression. Focusing on apoptosis-
related genes and their relationship with CD7, we observed an underexpression of most antiapoptotic
genes. Western blotting revealed increasing Akt and Bad phosphorylation, timely correlated with CD7
loss. This was shown to be phosphatidylinositol 3-kinase (PI3K)-dependent. Activation of PI3K/Akt
induced resistance to the apoptotic effect of interleukin-2 deprivation. We thus propose the following
model: HTLV-I infection induces a progressive decrease in CD3 genes expression, which eventually
abrogates CD3 expression; loss of CD3 is known to perturb calcium transport. This perturbation
correlates with loss of CD7 expression and induction of Akt and Bad phosphorylation via activation of
PI3K. The activation of the Akt/Bad pathway generates a progressive resistance to apoptosis, at a time
HTLV-I genes expression is silenced, thus avoiding immune surveillance. This could be a major event in
the process of the malignant transformation into ATLL.
128 Human domain antibodies against HIV-1 as
exceptionally potent cross-reactive neutralizers
Weizao Chen, PhD

CCR Nanobiology Program, NCI-Frederick, Bldg. 469, Rm. 144, Miller Drive, Frederick, MD 21702
HIV-1 has evolved to escape neutralization by antibodies generated by the immune system but
not by antibody fragments of smaller size that could be able to gain access to the highly guarded
conserved structures on the envelope glycoprotein (Env). Such small fragments targeting sterically
restricted regions on the Env could exhibit neutralization activity superior to larger antibodies as has
been demonstrated for Fab and scFv X5 which are significantly more potent than the full-size antibody
(IgG1 X5). We have hypothesized that further decreasing the size of the antibody fragments to the
smallest independently folded fragments, the antibody domains (about 10-fold smaller than an IgG,
but maintaining high binding affinity) could lead to exceptionally potent and broadly cross-reactive
neutralizers. To identify such fragments, we constructed a large (size 2.5 3 1010) highly diversified
library of human antibody variable domains (domain antibodies, dAbs), and used it for selection of
binders to conserved Env structures by panning sequentially against Envs from two different isolates,
one of which was complexed with CD4. The highest affinity binder, m36, neutralized all tested HIV-1
primary isolates from clades A, B, C, and D with an activity on average higher than that of C34,
a peptide similar to the fusion inhibitor T20 which is in clinical use, and that of m9 which is an
improved derivative of scFv X5. Increasing the size by joining m36 to Fc with hinge regions of varying
length or to Ch3 domains diminished its neutralizing activity likely due to the sterically restricted
nature of its epitope. M36 is the first representative of a novel class of potent and broadly cross-
reactive HIV-1 inhibitors based on human dAbs. It has potential as a candidate therapeutic, and as an
agent for exploration of the highly protected conserved Env structures with implications for the design
of small molecule inhibitors and elucidation of the mechanisms of virus entry into cells.
Key words: drug; domain antibody; entry; epitope; HIV-1; neutralization; steric restriction; therapeutic
129 Human Endogenous Retrovirus-K (HML-2) Elements
in the Plasma of People with Lymphoma and Breast
Cancer
Rafael Contreras-Galindo
University of Michigan, Department of Internal Medicine, Division of Infectious Diseases, 1150 West Me
Medical Center Dr., 5240 MSRBIII, Ann Arbor, MI 48108
Actively replicating endogenous retroviruses entered the human genome millions of years ago
and became a stable part of the inherited genetic material. They subsequently acquired multiple
mutations, leading to the assumption that these viruses no longer replicate. However, certain human
tumor cell lines have been shown to release endogenous retroviral particles. Here we show that RNA
from the human endogenous retrovirus HERV-K (HML-2), a relatively recent entrant into the human
genome, can be found in very high titers in the plasma of patients with lymphomas and breast cancer
as measured by either RT-PCR or Nucleic Acid Sequence Based Amplification (NASBA). Further, these
titers drop dramatically with cancer treatment. We also demonstrate the presence of reverse
transcriptase and viral RNA in plasma fractions that contain both immature and correctly-processed
HERV-K (HML-2) Gag and Envelope proteins. Finally, we show the presence of HERV-K (HML-2) virus-like
particles in the plasma of lymphoma patients using immunoelectron microscopy. Taken together, these
findings demonstrate that elements of the endogenous retrovirus HERV-K (HML-2) can be found in the
blood of modern-day humans with certain cancers.
130 Inhibition of HIV-1 Release by Cell Permeable
Peptides
Sarah I. Daniels, David A. Davis, and Robert Yarchoan

NIH, 9000 Rockville Pike, Bldg 10, Rm. 6N106, Bethesda, MD 20892-1868
Despite the effective use of highly active antiretroviral therapy to treat HIV infection viral,
resistance occurs over time and therefore new strategies to treat HIV infection re necessary. In the
process of developing peptide dimerization inhibitors of HIV-1 protease (Davis et al., Antiviral
Research, 2006, 72, 88), we found that certain peptides, originally designed as dimerization inhibitors,
were able to inhibit HIV-1 release. The peptide that was most effective at blocking dimerization, P27,
revealed a block in virus release of HIV-1 as measured by p24 in the media of treated H9 cells with
a corresponding increase in processed virus within the cells as evaluated by Western blot. P27 and
analogs also blocked the release of virus from transfected 293T cells. In the transfected 293T cells, the
virus did not accumulate significantly within the cells, but appeared to be degraded. The IC50 of
blockage of wt HIV-1 release from 293T cells by P27 was approximately 1.0 mM, and no significant
toxicity was seen at that concentration. P27 was designed to bind to the HIV protease dimer interface.
The potency of various analogs of P27 on inhibition of protease activity and dimerization did not
correlate to their potency on blocking virus release. Some peptides that were very weak at inhibiting
dimerization were still quite effective inhibitors of virus release. Also, different plasmid constructs
transfected in 293T cells showed that the inhibition of HIV-1 release was independent of the effects on
HIV protease. Moreover, peptides could inhibit the release of viral-like particles from cells transfected
with plasmid encoding only Gag. Characterization of P27 revealed that at least three of the four
domains are required to block the release of wild type and protease mutant virus. Further studies will
be required to understand the precise mechanism of action of these peptides and provide further
insights into the pathways important for viral release.
131 The gp41-derived immunosuppressive (isu) peptide
of HIV-1 modulates cytokine release and gene
expression in human immune cells
Magdalena Eschricht, Michael Lauck, Levent Akyüz, Rayk Behrendt,

Reinhard Kurth, and Joachim Denner
Robert Koch Institute, D-13353 Berlin, Germany
The transmembrane envelope protein gp41 of HIV-1 and a synthetic peptide corresponding to
a highly conserved domain of gp41, the so-called immunosuppressive (isu) domain, have been shown
to inhibit mitogen-triggered stimulation and to modulate cytokine production of human immune cells.
The expression of cytokines such as IL-10, IL-6, IL-8, RANTES, MCP-1, MCP-2, TNF-alpha, MIP-1alpha, MIP-
1beta, MIP-3 increased upon exposure to gp41 or the corresponding isu-peptide. In contrast, the
expression of IL-2 and MIG (CXCL-9) decreased. The modulation of cytokine expression induced by the
isu-peptide is similar to the changes in cytokine expression in HIV-1 infected individuals. To analyse
changes in gene expression in peripheral blood mononuclear cells (PBMCs) from healthy donors when
incubated with the isu-peptide, a microarray analysis (AB1700, 29098 human genes) was performed.
The microarray analysis confirmed the cytokine data and showed elevated expression of more than 400
genes, among them some of the above mentioned cytokines, MMP-1 (matrix metallopeptidase 1),
TREM-1 (triggering receptor expressed on myeloid cells 1) and others. By real time PCR studies
overexpression of these genes was confirmed and kinetics of the changes in gene expression were
analysed. The changes in gene expression were also shown at the protein level using ELISA and Western
blot analysis. In addition, preliminary data showed an elevated expression of FoxP3 and an increase in the
number of CD4+CD25+FoxP3+ regulatory T cells. These data indicate that gp41 and the corresponding
isu-peptide of HIV-1 modulate cytokine release and gene expression in normal human PBMCs.
132 Argotom-VAX: A Proposed Investigational HIV DNA
Vaccine Composed Of Selected Sequences Of Clade
B Gag, Pol, Nef, Tat, Vif, And Env Linked To Lysosomal
Membrane Associated Protein (LAMP)
William G. Hearl, PhD and J. Thomas August, MD (if available)

Immunomic Therapeutics, Inc., 9290 Gaither Road, Gaithersburg, MD 20877
HIV infection results in a progressive loss of CD4+ cells and the immune response of the host. The
aim of immune intervention in the chronic stage is to restore immunological competence of infected
individuals, reduce viral load, and retard CD4 cell loss. It has been shown by extensive animal and
human studies that the immunogenicity of the protein antigen can be enhanced by targeting the
antigen as a lysosome associated membrane protein (LAMP) chimera to the major histocompatibility
complex type 2 (MHC II) processing compartment of professional antigen presenting cells for
enhanced presentation to CD4 cells. Each of the three plasmids of the vaccine product will contain
three distinct domains of LAMP: (1) the highly glycosylated luminal domain with an N-terminal
endoplasmic reticulum translocation signal; (2) the transmembrane domain, and (3) the cytoplasmic
domain at the C-terminal. The antigen sequences are inserted between the luminal and
transmembrane domains. Studies of HIV-1 DNA immunogen formulations have described an HIV-1
p55Gag DNA construct with strong, broad and poly-functional cellular and humoral immune
responses in immunized mice when the Gag sequence was incorporated into the complete LAMP
cDNA sequence. The LAMP/Gag chimera with the complete LAMP protein co-localized with the MHC II
of transfected cells and elicited strong cellular and humoral responses in immunized mice compared
to DNA-encoding native Gag, with a 4-to 5-fold increase in CD8+ T-cell response and antibody titers
of .100,000.
The proposed investigational vaccine, Argotom-VAX, is composed of three closed, circular
supercoiled DNA plasmids containing the 8L-pCMV vector with three characterized nucleotide
sequences that encode HIV antigens: LAMP/gag, LAMP/pol, and LAMP/NTVE. The complete LAMP
protein will be used in each chimeric construct, designed for optimum protein expression and targeting
to the major histocompatibility II (MHC II) compartment of selected sequences of HIV-1 Clade B proteins,
p55 gag, pol, and a fusion of Nef-Tat-Vif-Env (gp41).
133 O-linked N-Acetylglucosaminylation Represses HIV-1
Replication and Sp1-Mediated Trans-Activation of
the HIV-1-LTR
Ramona Jochmann, Mathias Thurau, Elisabeth Naschberger, Susan Jung,

Elisabeth Kremmer, and Michael Stũrzl
Division of Molecular and Experimental Surgery, Department of Surgery, University of Erlangen-
Nuremberg, Schwabachanlage 10, 91054 Erlangen, Germany
The gene expression and replication of the human immunodeficiency virus-1 (HIV-1) is regulated
by the promoter/enhancer located in the U3 region of the proviral 5’-long terminal repeat (LTR). Binding
of the cellular transcription factor Sp1 to specific regulatory sites in the 5’-LTR is a key event in the
replication cycle of HIV-1. O-linked N-acetylglucosaminylation (OGlcNAcylation) is a metabolism-
associated highly dynamic and inducible posttranslational modification found on cytoplasmic and
nuclear proteins in eukaryotes, catalyzed by O-GlcNAc transferase (OGT) and reversed by OGlcNAc
hexosaminidase (O-GlcNAcase). Since the activity of Sp1 has been described to be regulated by
O-GlcNAcylation, we evaluated whether increased O-GlcNAcylation affects HIV-1 transcription. In the
present study we demonstrate that treatment of HIV-1-infected lymphocytes with the OGlcNAcy-
lation- enhancing agent glucosamine (GlcN) repressed viral replication in a dose dependent manner,
and that this effect is due to a decreased activity of the LTR promoter. Overexpression of OGT
specifically and dose dependently inhibited the activation of the LTR promoter containing the Sp1-
binding sites and strongly augmented the O-GlcNAcylation of Sp1. Knockdown of Sp1 by short hairpin
RNA blocked OGT repression of the HIV-1-LTR promoter, and depletion of OGT by RNA interference
completely restored the promoter activity. In contrast to the LTR promoter, OGT had no inhibitory
effect on the Sp1-dependent control promoter EF1-alpha. Finally, gel shift assays demonstrated that
O-GlcNAcylation does not alter the DNA-binding affinity of Sp1. From this study, we conclude that
OGlcNAcylation of Sp1 represses trans-activation of the HIV-1-LTR promoter. OGT does not act as
a general repressor of Sp1-triggered gene expression, but selectively inhibits the Sp1-mediated HIV-1-
LTR promoter activity. Thus, modulation of Sp1 O-GlcNAcylation may play a role in the regulation of
HIV-1 latency and activation, and may link viral replication to the metabolic state of the host cell.
134 Cell-cycle dependent HIV-1 killing is mediated
through the viral protease
Kyeongeun Lee, Taichiro Takemura, Yujing Xue, Alok Mulky, and

Vineet N. KewalRamani
HIV Drug Resistance Program, National Cancer Institute, Frederick, MD 21702, USA
HIV-1 infection in vivo is characterized by the profound, cytopathic depletion of CD4+ helper T
cells. Different mechanisms have been proposed to underlie HIV-1 cell killing. High multiplicity HIV-1
infection is thought to kill cells via toxic levels of unintegrated vDNA or lethal integration events.
Killing by low multiplicity HIV-1 infection is thought to require the accumulation of toxic gene
products. Using an HIV-1 vector that lacks several cytotoxic genes (vif, vpr, env) and can be stably
transduced, we observed killing of human cells at infection frequencies as low as one hit per cell.
Interestingly, slowly dividing cells were less rapidly killed after infection. Consistent with this
observation, cells growth arrested with anti-proliferative drugs were not killed by HIV-1 vector
infection. However removal of drugs and relief of the cell-cycle block rapidly led to death in the
infected cells. HIV-1 vector expression was found to be required for the killing. A comparison of cells
stably or acutely infected with the HIV-1 vector revealed higher viral protein expression during acute
infection and more efficient proteolysis of Gag. Treatment of cells with HIV-1 PR inhibitors (PIs)
alleviated killing during acute infection but did not reduce overall viral protein expression. PIs also
protected initially growth-arrested HIV-1 vector infected cells after relief of the cell-cycle block. By
contrast, HIV-1 vectors encoding drug-resistant PR killed dividing cells in the presence of PIs.
Collectively, our data indicate that HIV-1 PR expressed from a single provirus is a cytotoxic hazard to
proliferating cells.
135 Dramatic enhancement of uptake and trafficking of
HIV-1 Tat protein via modulation of endocytosis
pathways
Guan-Han Li, Wenxue Li, Yan Huang, and Avindra Nath

Johns Hopkins School of Medicine, Dept of Neurology, 600 N Wolfe St, Baltimore, MD 21287
HIV-1 Tat protein can be secreted from infected cells and taken up by other target cells. Tat uptake
and trafficking has been widely investigated. This property is also exploited for drug delivery. We
initially confirmed that Tat mainly entered Hela cells by clathrin-dependent endocytosis and was
degraded in lysosomes, whereas chloroquine could inhibit the degradation and enhanced its
trafficking to the nucleus. Interestingly, a similar effect was noted in presence of chlorpromazine (CPZ).
Most likely, the uptake of Tat was shifted to caveolar endocytosis after the clathrin pathway was
blocked by CPZ. However, the LTR transactivation of Tat could be enhanced by at least 50 fold when
a liposome reagent was introduced into the Tat delivery system. Further evidence showed that liposome
reagents could switch the Tat uptake from clathrin-dependent endocytosis to lipid raft pathway and
bypass the degradation in lysosomes. This was supported by both pharmacological assays using
inhibitors of caveolae-mediated endocytosis (MbCD, etc.) or microtubules (cytochalasin D &
lutrunculin A) and co-localization using Tat101venus and immunstaining with anti-clathrin1, anti-
caveolin1, anti-EEA1, anti-Lamp1 or anti-Rab7. Subsequently, chemical molecules carrying ‘‘+’’ or ‘‘2’’
charges were tested for blocking the Tat uptake facilitated with the liposome reagent, including
macromolecules (e.g., heparin, polybrene) and small molecules (e.g., spermine). Both ‘‘+’’ and ‘‘-’’
charged molecules could inhibit the uptake of Tat, but the ‘‘+’’ molecules actually attached to the
membrane carrying ‘‘–’’ charges and the ‘‘–’’ molecules bound to the Tat-liposome complex with ‘‘+’’
charges. However, the Tat-liposome complex was probably formed by hydrophobic bonds linking
liposome’s lipid to Tat hydrophobic domain because both Tat and the liposome reagent carried
‘‘+’’charges. A peptide containing partial hydrophobic domain of Tat (CFITKGLGISYGRKK) could
competitively inhibit the uptake of Tat-liposome complex. In conclusion, the liposome reagent could
facilitate Tat protein to traverse the membrane by changing endocytosis pathways and significantly
enhance its delivery to the nucleus in a lysosome-independent manner. These observations have
important implications for elucidating the mechanism of Tat uptake and trafficking and exploring drug
delivery.
Key Words: HIV-1 Tat; liposome; uptake
136 Increased IL-15 production is associated with
higher infection of memory CD4 T cells during acute
SIV infection
Joseph Mattapallil
F. W. Herbert School of Medicine, USUHS, 4301 Jones Bridge Road, Bethesda, MD 20814
Acute SIV infection is characterized by explosive infection of memory CD4 T cells in peripheral and
mucosal tissues. Interestingly, relatively few memory CD4 T cells in either the mucosa or periphery of
SIVinoculated rhesus macaques are infected until as late as day 7–8 after challenge. However, by day
10 pi most of the memory CD4 T cells in both tissues are infected and carry viral DNA. The rapid pace
with which infection expands within 2–3 days to encompass virtually the entire memory CD4 Tcell
compartment suggests significant alterations in the susceptibility of memory CD4 T cells to infection
during this period. The mechanism(s) underlying this increased permissiveness for infection is not
known. Here we show that IL-15 secretion significantly correlates with the upregulation of CD4
expression on memory CD4 T cells leading to their increased permissiveness to SIV infection. Activation
and proliferation of memory CD8, but not memory CD4 T cells, preceded the amplification of viral
infection that significantly correlated with IL-15 production. Though memory CD4 T cells did not
express normal activation markers they displayed a significant upregulation in the density of CD4
expression between day 7 and 10 pi that correlated with increased infection of these cells. Culture of
purified populations of CD4 T cells with IL-15 and/or SIV upregulated the expression of CD4 on these
cells, and was accompanied by a significant increase in the level of infection in these sorted cells. Our
results demonstrate that IL-15 contributes to the increased susceptibility of memory CD4 T cells to SIV
during the early phase of acute SIV infection.
137 Presence and role of HLA-C in HIV-1 infection
Andrea Matucci,1 Paola Rossolillo,1 Marco Turci,1 Pierpaolo Racchiolli,1

Antonio G. Siccardi,2 Alberto Beretta,2 and Donato Zipeto1
1
Section of Biology and Genetics, University of Verona, Italy and 2San Raffaele Scientific Institute,
Milan, Italy
Background: A whole genome association study reported a SNP at -35Kb from the HLA-C gene
strongly associated to HIV-1 viral set point and HLAC expression level (Fellay J. et al., 2007). HLA-C is
not down-modulated by HIV-1 Nef and can be specifically incorporated in viral membrane enhancing
infectivity and resistance to neutralizing antibodies (Cosma A. et al., 1999). We investigated the role of
HLA-C in modulating HIV-1 infectivity using cell fusion and pseudovirus infection models and the
interaction between HLA-C and Env at membrane level and in purified fusion complexes.
Methods: Human cell lines expressing different HIV-1 gp120/gp41 were specifically silenced for
HLA-C expression. Cells or pseudoviral particles were used for fusion and single-cycle infection analysis
with TZM-bl cells. Fusion efficiency and viral infectivity were compared. Fusion complexes from fusing
cells were purified and the molecular proximity of HLA-C and Env was analyzed by using BRET2.
Results: The absence of HLA-C significantly decreased the fusion efficiency of cells expressing
different R5 and X4 tropic gp120/gp41s. Similarly, pseudovirus infectivity was significantly reduced if
they were produced on HLA-C silenced 293T cells. The X4 tropic NDK Env was insensitive to HLA-C at
higher infectious doses. VSV-G pseudovirus used as control was not sensitive to HLA-C presence. HLA-
C could be detected associated to gp120 in cells taken before fusion, albeit a stronger association was
evident during fusion process with target cells.
Conclusions: The co-expression of HLA-C with X4/R5 HIV-1 Env increases the fusogenicity of cell
lines and the infectivity of pseudotyped viruses. This data point to a specific interaction between HLA-
C and gp120 on the cell surface membrane. HLA-C and gp120 increase their association during fusion
complex formation. This interaction can stabilize Env trimers, increasing the kinetic of conformational
changes or the exposure of the receptor and/or coreceptor binding site favoring membrane fusion.
Inhibiting the HLA-C/Env interaction might be of great importance in reducing virus infectivity and
become the rationale for the development of new inhibitors of HIV-1 entry.
138 Prevalence of Human Papillomavirus (HPV) Infection
among HIV-positive women in the Pre-HAART and
HAART era in a Nigerian Clinic
Dr. Olanrewaju Onigbogi

Department of Community Medicine, University College Hospital, Ibadan
Objectives: The prevalence of HIV infection has been on the increase in Nigeria in recent times.
HIV-positive patients frequently have anogenital malignancies due to HPV. HAART was introduced in
Anti-retroviral (ARV) centers in Nigeria in the year 2002. The aim of the study is to determine trends in
incidence of anogenital malignancies among HIV-positive women undergoing treatment in the clinic
in the pre-HAART and HAART era.
Methods: A retrospective review of 541 case notes of HIV-positive female patients from January
1999 to December 2004 were analyzed by utilizing an on-going observational database at the ARV center.
Rate ratios, comparing incidence rates (number of malignancies per 1000 person years) were calculated.
Results Obtained: Twenty-four (4.43%) of the patients had one form of anogenital manifestation
of HPV or the other. The incidence rate for HPV rose from 2.28 in the pre-HAART era to 6.40 in the
HAART era (Rate ratio = 3.15; 95% confidence interval (CI) = 1.31 – 7.44; P = 0.0002).
Conclusions: There has been a significant rise in the incidence of HPV since the introduction of
HAART. This may be due to the longer survival of HIV-infected patients, surpassing the latency period
for the anogenital malignancies. Care providers should be more vigilant for HIV-associated
malignancies as patients live longer in this part of the world.
139 HIV-2 Capsids Distinguishing High and Low Virus
Load Patients in a West African Community Cohort
Clayton Onyango, 1 Aleksandra Leligdowicz, 1,2 Masaru Yokoyama, 4

Hironori Sato,4 Haihan Song,3 Emi Nakayama,3 Tatsuo Shioda,3 Assan Jaye,1
Hilton Whittle,1 Sarah Rowland-Jones,1,2 and Matthew Cotten1
1
MRC Laboratories, The Gambia; 2WIMM, John Radcliffe Hospital, Oxford, OX3 9DS, United Kingdom;
3
Department of Viral Infections, Research Institute for Microbial Diseases, Osaka University, Japan; and
4
Laboratory of Viral Genomics, Center for Pathogen Genomics, National Institute of Infectious Diseases,
Tokyo, Japan
HIV-2 causes AIDS similar to HIV-1; however, most HIV-2 infected patients show no disease and
have low plasma virus load (VL). A detailed sequence analysis of HIV-2 p26 capsid (P26) variation in
a community cohort was undertaken with the primary objective of detecting p26 variation that
correlated with virus load.
Activities: Sensitive methods were developed to generate virus sequences from high and low
virus load subjects, providing information from both healthy and progressing HIV-2+ patients.
Results: Three amino acid polymorphisms in p26 were identified that correlated significantly with
VL. Proline residues at the three sites (PPP) were found more frequently in lower VL subjects while p26
with non-proline residues at all three sites (non-PPP) were isolated more frequently from high VL
subjects. Structural modeling of these capsid variants revealed an increase in capsid dimer binding
energies associated with the high VL (non-PPP) variants. In vitro virus replication supported the
conclusion that these residues influence TRIM5a interaction, with PPP variants having increased
susceptibility to TRIM5a restriction. The cellular immune responses to HIV-2 antigens in PPP patients
were significantly higher than in non-PPP patients, indicating that capsid phenotype may also
influence antigen presentation.
Lessons: These results demonstrate that HIV-2 replication in vivo can be linked to p26 variation
and provide a new basis for understanding HIV-2 pathogenesis.
140 Risk factors associated with a high chance of mother
to child transmission (MTCT) of HIV: a retrospective
study of HIV-exposed infants/children in Kitwe, Zambia
Gilbert Siame
Zambia Rural Healthcare Outreach Services (ZARHOS), Kitwe, Zambia C/O Theresa Chalwe, Jambo Flats
No.11, Jambo Drvie, Parklands, Kitwe, Zambia
Background: In the absence of intervention to curb transmission to children, studies suggest that
the risk of transmission from an HIV infected mother to her child is 25–35%. In Zambia, about 112 new
HIV infections in babies occur per day, of the 520,000 babies born annually.
Objective: To identify risk factors that make a mother more likely to transmit HIV to her baby, and
come up with recommendations to reduce MTCT of HIV and improve infant survival.
Method: This was a retrospective study in which 100 case records of HIV exposed babies were
reviewed. Polymerase Chain Reaction (PCR) to detect HIV DNA had been done in all cases.
Results: From the 100 cases reviewed,13% of the babies were HIV positive. Of these positive
babies, 69% were still b/feeding beyond 9 months. 54% were b/fed exclusively, while 15% were having
mixed feeding. 62% of the mothers to these positive babies did not get any intervention to prevent
MTCT(PMTCT). 23% of their mothers got SD NVP whereas 15% had AZT+SD NVP.As for the babies, 31%
received SD NVP, while 7% got AZT+SD NVP. 85% of the babies’ mothers experienced an obstetric
event with bleeding, with 31% of the positive babies asphyxiated. There was h/o untreated STI in 15%
of the mothers, with more than half of the mothers (54%) having booked late in pregnancy. 31% of the
mothers had CD4 counts ,200 cells/ml around time of conception, with 0% having done viral loads.
Conclusion:
1) Vertical transmission (MTCT) is still the overwhelming source of HIV infection in babies, with low
maternal immunity/poor clinical status, prolonged b/feeding, obstetric trauma, playing a major
contributory role.
2) Women should be encouraged to do viral loads/CD4 counts prior to conception, and avoid
pregnancy if viral loads .50 000 copies/ml or CD4,200 cells/mL.
3) Need to avail women in developing countries with appropriate and affordable alternatives to
b/feeding.
4) Skilled health workers should attend to HIV positive pregnant women in labour to minimize adverse
obstetric events such as trauma, hemorrhage, birth asphyxia, stillbirth.
5) PMTCT programs should be part of a broader health care strategy; they need to be holistic, requiring
an expanded and strengthened prenatal, delivery and postnatal care programme.
6) Policy makers should address the cultural, legal and economic factors that make girls and women
especially vulnerable to HIV infection.
141 Type C Coping and Alexithymia are Associated
Differentially with Specific Immune Mechanisms
(Interleukin-6 and Beta-Chemokine Production)
Linked to HIV Progression
Lydia R. Temoshok,1 Rebecca L. Wald,1 Stephen J. Synowski,1 Alfredo Garzino-

Demo,1 and James Wiley2
1
Institute of Human Virology, University of Maryland School of Medicine, Baltimore MD, 2Public
Research Institute, San Francisco State University, San Francisco CA
We hypothesized that b-chemokines MIP-1a/b which inhibit HIV entry into CD4+ T lymphocytes
via the CCR5 coreceptor, and/or proinflammatory cytokines (Particularly IL-6) which activate HIV
replication, may be mechanisms underlying our previously reported finding that higher Type C coping
predicted HIV progression. The present study examined baseline relationships between Type C coping
(which allows unrecognized, unexpressed emotions to remain as chronic stressors) and the adjacent
personality construct of alexithymia (emotion processing/regulation deficits) and these immune
mechanisms linked to HIV progression. Participants were 200 HIV+ outpatients (92% African-American,
53% male, mean age 45). Type C was assessed by the Vignette Similarity Rating Method; alexithymia by
the Toronto Alexithymia Scale. In vitro production of IL-6 and MIP-1a/b was measured in response to
Candida, PHA, and the HIV core protein p24. A Stimulation Index was defined as antigen-stimulated
chemokine/IL-6 production compared to unstimulated controls. In regression analyses with
independent variables Type C and the total alexithymia score, controlled for age, CD4+ count,
antiretroviral medications, and adherence, higher Type C coping was significantly associated with IL-6
production stimulated by Candida (b =.207, P , 01), PHA (b = .182, P , 02), and p24 (b = .138, P =
.075). Regression analysis with the same IVs revealed a significant negative association for MIP-1a; and
alexithymia (b = 2.196, P = .008). To summarize: higher Type C coping is associated differentially with
higher IL-6 production, which amplifies HIV replication; while higher alexithymia is associated with
lower production of the anti-HIV chemokine MIP-1a, suggesting a role for psychoneuroimmunological
processes in HIV progression.
142 HIV Frequently Elicits Mucosal and Plasma Env-
Specific IgA With a Rapid Initial Decline In Acute
Infection
Nicole L. Yates, Judith Lucas, Rob Parks, Vicki C. Ashley, Glenn Overman,
David C. Montefiori, Kent J. Weinhold, Robin Shattock, Myron Cohen,
Barton F. Haynes, and Georgia D. Tomaras, the CHAVI 001 Team
Duke University Medical Center, MSRBII, Research Drive, Durham, NC 27710
An optimal HIV vaccine would elicit protective immune responses, preferably neutralizing
antibodies, at mucosal surfaces in order to prevent infection. One key to eliciting vaccine-induced
antibody responses at mucosal surfaces is to understand what antibodies are present at these sites
soon after transmission during acute HIV infection (AHI). However, past attempts to detect HIV-specific
IgA in mucosal specimens have been largely unsuccessful. The goal of this project is to characterize the
binding antibody reponses elicited during AHI in plasma and mucosal (semen and cervicovaginal
lavage (CVL)) samples compared to chronic infection with respect to antibody isotype, specificity, and
timing in the CHAVI 001 acute infection cohort. To determine the presence of binding antibodies to
HIV envelope proteins (gp140 consensus oligomers and recombinant gp41) and p55 gag protein,
a standardized colorimetric ELISA as well as a customized Luminex assay were performed. Luminex is
a bead-based flow cytometric assay that we customized to create a highly sensitive multiplexed assay
for determining HIV-specific isotypes in plasma and mucosal specimens. Our results are the first to
demonstrate the presence of HIV-specific IgA in approximately 88% of mucosal samples obtained
during AHI, whereas approximately 33% of samples from chronically infected patients were positive
for HIV-specific IgA. Each of the AHI patients had env-specific mucosal IgA, some of which mapped to
the immunodominant region of gp41. High levels of HIV-specific IgG were also observed in the acute
and chronic mucosal samples while HIV-specific IgM was undetectable. Furthermore, HIV env-specific
plasma and mucosal IgA levels rapidly declined over time. In addition, in approximately 40% of AHI
patients, systemic levels of p55 gag-specific IgA rose over the course of the infection despite the
decrease in env-specific IgA. The rapid decline in HIV env-specific IgA suggests mucosal B cells may
lose the capacity for env-specific IgA production during the course of AHI. Furthermore, the concurrent
rise in systemic gag-specific IgA levels over time may be a reflection of HIV env-mediated differential
regulation of IgA production in activated mucosal B cells. Future work includes functional studies to
determine the role of HIV-specific IgA in AHI versus chronic infection as well as more epitope mapping
to determine the specificity of isotypes in mucosal samples. These results have important implications
in our understanding of how HIV infection regulates mucosal B cell responses and the barriers that
need to be overcome by an HIV vaccine.
143 Broad Spectrum Neutralizing Antibodies Against
HIV-1 Elicited by Immunizing with Fusion
Complexes
P. Rossolillo,1 A. Matucci,1 P. Racchiolli,1 L. Mainetti,2 S. Dispinseri,1

G. Scarlatti,2 and D. Zipeto1
1
University of Verona, Italy; 2DIBIT-HSR, Milan, Italy
Background: The development of neutralizing antibodies against HIV-1 is of pivotal importance for
the development of an AIDS vaccine. We immunized mice with fusion complexes and elicited antibodies
with neutralizing activity against heterologous HIV-1 isolates (Zipeto et al., Microb Infect 2006).
Methods: We prepared murine hybridomas from mice whose sera showed the highest
neutralizing activity. Their supernatants were tested for reactivity against cells expressing CD4 and
CCR5, gp120/41, and the gp120/CD4 complex formed by capture ELISA. Hybridoma antibodies with no
reactivity against HIV-1 receptors were selected. IgGs were purified and tested for their neutralizing
activity (1–5 mg/mL) using both the TZM-bl neutralization assay (Li M. et al, J Virol 2005) and pNL4-
3.Luc.R-E- based pseudoviruses on U87R5 cells with the standard group B Env panel. Specificity was
tested against the VSV-G envelope protein. The neutralizing activity of selected antibodies was
confirmed using the PBMC-based neutralization assay (Polonis VR et al, Virol 2008).
Results: We screened 150 different hybridoma groups; 8% showed a neutralizing activity higher
than 40% and 1 between 50 and 80% against the different pseudoviruses tested, similar or higher than
the Tri-mAb positive control. Cells from this hybridoma group were cloned; 7% showed a neutralizing
activity higher than 40% and 2% higher than 50%. Among the latter as many as 44% showed
a neutralizing activity higher than 75% against the different pseudoviruses tested.
Conclusions: Hybridomas produced from mice immunized with fusion complexes secrete
antibodies neutralizing a panel of HIV-1 clade B Envs. We are screening monoclonal antibodies for their
broad spectrum neutralizing activity against HIV-1. The isolation of such monoclonal antibodies will be
of great interest for the development of an AIDS vaccine.
144 In vivo and ex-vivo proteomics for target discovery
in cancer
Giuliano Elia
University College Dublin Conway Institute, Belfield, Dublin 4, Ireland
There is an urgent need for more selective anticancer drugs. Targeted delivery of bioactive
molecules to the tumor environment by means of binders (such as recombinant human antibodies
and their fragments) specific to tumor-associated markers is a promising strategy towards this goal.
This approach crucially relies on the availability of good quality, tumor-specific antigens and suitable
high affinity ligands. Target proteins located around tumor blood vessels and in the stroma appear to
be particularly suited for targeted anticancer strategies in view of their abundance, stability and
accessibility by intravenously administered biopharmaceuticals.
We have established ‘‘2D peptide mapping’’ as a new method for selective labeling, separation,
relative quantification and identification of membrane proteins. This method is based on biotinylation
of cell surface proteins, purification on streptavidin columns, elution, digestion with trypsin and
peptide separation through reverse-phase HPLC. MALDI mass spectrometric profiles of each fraction
can be displayed in two-dimensional maps that allow the immediate comparison of related samples
and quantification of significant differences. We have applied this method to an in vivo proteomic
approach for discovery of tissue specific targets accessible from circulation. We performed terminal
perfusion of tumor-bearing rodents with a biotin derivative, which cannot diffuse through biological
membranes. Comparative proteomic analysis by 2D peptide mapping revealed candidate targets
differentially expressed in normal organs vs. experimental tumors.
Finally, we have performed the ex vivo perfusion and biotinylation of whole, surgically resected
human kidneys with tumor to gain information about accessible and abundant antigens that are
overexpressed in human cancer. This procedure led to the selective labeling with biotin of vascular
structures. Biotinylated proteins were purified on streptavidin resin and identified using 2D peptide
mapping, revealing 637 proteins, 184 of which were only found in tumor specimens and 223 of which
were only found in portions of normal kidneys. Immunohistochemical and PCR analysis confirmed that
several of the antigens identified are indeed preferentially expressed in tumors.
145 The V1/V2 Loop Region of Simian-Human
Immunodeficiency Virus Envelope gp120 is the
Major Determinant of the Strain Specificity of the
Neutralizing Antibody Response
Melissa E. Laird, Tatsuhiko Igarashi, Malcolm A. Martin, and

Ronald C. Desrosiers
Harvard Medical School
Plasma samples from individuals infected with human immunodeficiency virus type 1 (HIV-1) are
known to be highly strain-specific in their ability to neutralize HIV-1 infectivity. Such plasma samples
exhibit significant neutralizing activity against autologous HIV-1 isolates but typically exhibit little or
no activity against heterologous strains. Monkeys infected with the simian-human immunodeficiency
virus (SHIV) clone DH12 generated antibodies that neutralized SHIV DH12 but not SHIV KB9.
Conversely, antibodies from monkeys infected with the SHIV clone KB9 neutralized SHIV KB9 but not
SHIV DH12. To investigate the role of the variable loops of the HIV-1 envelope glycoprotein gp120 in
determining this strain specificity, variable loops 1 and 2 (V1/V2), 3 (V3) or 4 (V4) were exchanged
individually or in combination between SHIV DH12 and SHIV KB9. Despite the fact that both parental
viruses exhibited significant infectivity and good replication in the cell lines examined, three of the ten
variable loop chimeras exhibited such poor infectivity that they could not be used further for
neutralization assays. These results indicate that a variable loop that is functional in the context of one
particular envelope background will not necessarily function within another. The remaining seven
replication-competent chimeras allowed for unambiguous assignment of the sequences principally
responsible for the strain specificity of the neutralizing activity present in SHIV-positive plasma.
Exchange of the V1/V2 loop sequences conferred a dominant loss of sensitivity to neutralization by
autologous plasma and gain of sensitivity to neutralization by heterologous plasma. Substitution of V3
or V4 had little or no effect on the sensitivity to neutralization. These data demonstrate that the V1/V2
region of HIV-1 gp120 is principally responsible for the strain specificity of the neutralizing antibody
response in monkeys infected with these prototypic SHIVs.
146 a4b7: A Newly Discovered Receptor for HIV
Anthony S. Fauci, MD
National Institute of Allergy and Infectious Diseases, NIH, Bethesda, MD
Infection with HIV-1 results in the dissemination of virus to gut-associated lymphoid tissue
(GALT). Subsequently, HIV mediates massive depletion of gut CD4+ T cells, which contributes greatly
to HIV-induced immune dysfunction. The migration to and retention of lymphocytes in GALT is
mediated by integrin a4b7. We have demonstrated that the HIV envelope protein gp120 binds to an
activated form of a4b7. This interaction is mediated by a tripeptide in the V2 loop of gp120,
a tripeptide that mimics structures presented by the natural ligands (MadCAM, VCAM-1, and
fibronectin) of a4b7. Engagement by gp120 of a4b7 on CD4+ T cells results in rapid activation of
LFA-1, the major integrin involved in the establishment of virological synapses, which facilitate
efficient cell-to-cell spread of HIV. These findings may have important implications in the further
delineation of the pathogenic mechanisms of HIV infection.
147 Hepatitis C and cancer
Michael Houghton, PhD

Epiphany Biosciences Inc., San Francisco
The hepatitis C virus (HCV) is a positive-stranded RNA virus classified within the Flaviviridae
family. It infects 3-4 MM individuals in the USA alone and an estimated 170 MM worldwide. Around
75% of acute infections develop into life-long infections which are associated with varying levels and
types of liver and other diseases. While the majority of infected individuals only have mild forms of
hepatitis, some can develop chronic lobular hepatitis that then proceeds sequentially through
chronic active hepatitis, liver fibrosis, liver cirrhosis and primary hepatocellular carcinoma. The latter
is always preceded by cirrhosis suggesting that the continually-inflamed liver may itself be a direct
cause of tumorigenesis .However, certain HCV gene products may also play a role since the
nucleocapsid and nonstructural proteins 3 & 4 have been shown to induce tumorigenesis when
overexpressed in various cellular and animal models. HCV-associated extrahepatic diseases
include mixed cryoglobulinemia, membranoproliferative glomerulonephritis, porphyria cutanea tarda
and there is some association with non-Hodgkin’s B cell lymphoma. The latter may be mediated by the
engagement of the HCV receptor, CD81 by envelope glycoprotein 2 within circulating HCV virions.
Many co-factors of HCV-associated disease are known including gender (with the incidence of
hepatocellular carcinoma being lower in females), alcohol use, duration of infection and co-infection
with other hepatotropic viruses as well as co-infection with HIV. Individuals co-infected with HIV & HCV
often develop severe liver disease.
Vaccine strategies to prevent the development of persistent infection are now looking promising.
Vaccines eliciting cross-neutralising antibodies or cross-reacting cellular immune responses in non-
human primates are able to prevent chronic infection following experimental challenge with
heterologous viral strains thus mirroring the correlates of protection in humans that are able to
spontaneously eradicate acute HCV infection.
Standard-of-care therapy comprises pegylated alpha-interferon along with ribavirin, a mixture
that is thought to have direct antiviral and immunomodulatory activity. Patients with chronic hepatitis
and even liver cirrhosis can be normalized by this therapy with virus eradication, although 12 months
of treatment are required for the common genotype 1 infections and even then, less than 50% of
patients are permanently cured. However, progress is occurring through the use of more specific
antivirals that inhibit the viral serine protease, the viral replicase and other targets like nonstructural
protein 5a that plays an essential role in virion production. Therefore, we can anticipate a combination
therapy to emerge within the next decade that will cure the large majority of HCV patients and
a vaccine that could prevent the chronic sequelae of the estimated 25,000 new infections still
occurring annually in the USA alone.
148 Towards a genetics of cancer resistance
George Klein
Microbiology and Tumor Biology Center (MTC), Karolinska Institute, Stockholm, Sweden
Approximately two of three humans never develop cancer. Not even the majority of heavy
smokers get cancer. Are there cancer resistant genotypes?
Mice can be readily selected for low cancer incidence by selective breeding. Feral-derived mice,
e.g. M.spretus are highly cancer resistant.
Among known cancer protection mechanisms, DNA repair, the stringency of epigenetic
imprinting, apoptotic propensity and immune response to virus associated tumors are influenced by
genetic variation. Contactual inhibition of tumor by adjacent normal cells may reflect a form of
microenvironmental control that could be responsible for preventing the cancerous progression of
initiated cells, the growth of disseminated and dormant tumor cells, and the progression on incipient
neoplastic foci. This mechanism has not been studied for genetic variation. A high throughput
microwell system has been developed in our laboratory that may be suitable for such studies.
In the field of somatic cell genetics, our studies on monochromosomal microcell hybrids have
revealed the existence of ‘‘asymmetric’’ tumor suppressor genes that can inhibit tumor growth in vivo
but do not influence in vitro growth. This study may open the way towards the exploration of cellular
receptors for contactual control.
149 The role of the microenvironment in EBV associated
lymphoid malignancies
Eva Klein
Department of Microbiology, Tumor and Cell Biology, Karolinska Institute, 17177 Stockholm Sweden
Depending on the differentiation and maturation of EBV-carrying cells, virally encoded proteins
are expressed in various combinations. These proteins determine the fate of the viral genome
harbouring cells. Virus transformed B lymphocytes - lymphoblastoid cell lines- LCL- express six virally
encoded nuclear and three surface localised proteins. This phenotype is encountered only in B
lymphocytes and it is associated with cell proliferation. It is usually referred to as Type III EBV
expression or growth transformation program. Such cells are readily recognized by the immune
response. Their proliferation therefore is inhibited in healthy individuals. In immunosupressed patients
this inhibition is impaired and EBV associated haematological malignancies may arise.
The presence of the EBV genome in lymphocytes with a more restricted viral protein expression,
like the type II pattern that lacks the nuclear protein EBNA-2, does not induce proliferation. However it
may affect the behaviour of the cell within the dynamics of the immune system. Such cells may avoid
apoptosis, they may induce an enrichment of inflammatory cells and/ or may respond to intercellular
contacts or to cytokines with proliferation. This may lead to malignancy.
There is evidence for such mechanisms in EBV associated Hodgkińs lymphoma and nasal
NK lymphoma.
150 HIV and cancer: current trends and insights
Eric A. Engels
National Cancer Institute, 6120 Executive Blvd, Room 7076, Rockville, MD 20852
HIV-infected individuals have an elevated risk for cancer. This high risk partly arises from HIV-
induced immunosuppression, and partly from a high prevalence of cancer risk factors such as tobacco
use and infections with oncogenic viruses. HAART, widely available since 1996, can result in substantial
improvements in immune status and has led to declines in AIDS incidence and mortality.
Monitoring of cancer trends in HIV-infected persons in the U.S. has revealed population-level
declines in the incidence of Kaposi sarcoma and non-Hodgkin lymphoma, to some extent attributable
to introduction of HAART in 1996. Of interest, Hodgkin lymphoma incidence has risen during the
HAART era. Hodgkin lymphoma risk is related to CD4 count in a non-linear manner, declining at the
lowest CD4 counts, and the rise in Hodgkin lymphoma incidence may reflect effects of HAART (i.e.,
a shift in immune function to a mid-range with highest Hodgkin lymphoma risk). Among cancers
caused by human papillomavirus, cervical cancer incidence has remained steady, but incidence of anal
cancer has increased over time.
Lung cancer is the most common non-AIDS-defining cancer in HIV-infected persons. This reflects
very high smoking rates, but lung cancer incidence appears greater than can be attributed to smoking
alone. The high risk of lung cancer in HIV-infected persons may be due to repeated lung infections or
chronic pulmonary inflammation, in concert with tobacco use. Incidence of non-melanoma skin
cancers is also elevated. Data on basal cell and squamous cell skin carcinomas are limited, but risk is
strongly increased for Merkel cell carcinoma and sebaceous carcinoma, two rare types of skin cancer. A
novel polyomavirus was recently discovered in Merkel cell carcinoma.
Trends in cancer incidence in HIV-infected individuals reflect the effects of HAART as well as other
factors. Studies of cancers that arise at increased frequency in this population may reveal novel
biological insights.
151 MicroRNAs and the biology of KSHV infection
Don Ganem
HHMI, UCSF San Francisco, CA 94143
Pre-miRNAs as part of its latency program; these are processed to yield 17 mature viral miRNAs.
The functions of most of these miRNAs are unknown. To address this issue, we have developed an
expression- based screen for candidate cellular mRNA targets of each of these miRNAs. Here, we report
the results for one such miRNA, miR-K5. Following ectopic expression of individual miRNAs, expression
profiling was carried out, looking for mRNAs whose abundance declines in the presence of the miRNA;
in addition, we transfected latently infected cells with antisense antagonists of each miRNA, then
looked for transcripts whose level rose in response to miRNA inhibition. Transcripts passing both sets
of tests were then examined for seed sequence homologies in their 3’UTR. For miRK5, the leading
candidate to emerge was the mRNA encoding the protein BCLAF1. This was validated as a target by
extensive formal genetic tests. Additional testing revealed that BCLAF1 was also targeted by at least 2
other KSHV miRNAs, pointing to an important role in the life cycle. Inhibition of BCLAF1 by siRNA
phenocopied the expression of KSHV miRNAs, and resulted in enhanced spontaneoud lytic
reactivation. We conclude that KSHV miRNA-mediated modulation of BCLAF1 promotes the
reversibility of latent infection, thereby preventing latency from becoming a dead-end pathway
from which virus cannot escape.
152 HIV-ASSOCIATED LYMPHOMAS — Focus on unusual
lymphomas occurring specifically in HIV-infected
patients
Antonino Carbone
Fondazione IRCCS Istituto Nazionale Tumori – Via G. Venezian, 1 – 20133 Milano, Italy
Lymphomas that develop in HIV-positive patients are predominantly aggressive B-cell

lymphomas. These disorders include the same lymphomas that develop sporadically in the absence
of HIV infection and those seen much more often in the setting of HIV infection. The most common
HIV-associated lymphomas are Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) (also
involving the central nervous-system-CNS). Classic Hodgkin lymphoma (HL) is also increased in the
setting of HIV.
Lymphomas occurring specifically in HIV positive patients include primary effusion lymphoma
(PEL) and its solid variants, plasmablastic lymphoma of the oral cavity type and lymphoma associated
with HHV8-related multicentric Castleman disease. These lymphomas together with BL and
immunoblastic lymphoma subtypes with plasmacytoid differentiation carry EBV infection and display
a phenotype related to plasma cells.
EBV is identified in the neoplastic cells of approximately 40% of HIV-associated lymphomas, but
the detection of EBV varies considerably with the site of presentation and histological subtype. EBV
infection occurs in 80-100% of primary CNS lymphomas and PELs, 80% of DLBCLs with immunoblastic-
plasmacytoid features, and 30-50% of BL-plasmacytoid. Nearly all HL cases in the setting of HIV
infection are associated with EBV. KSHV/HHV8 is specifically associated with PEL, which usually occurs
in the late stages of disease, in the setting of profound immunosuppression.
The current knowledge about HIV-associated lymphomas can be summarized in the next essential
points: 1) lymphomas specifically occurring in patients with HIV infection are closely linked to other viral
diseases; 2) most of these lymphomas exhibit plasmablastic differentiation; and 3) among HIV-associated
lymphomas the incidence of classic HL is increasing in the setting of improved immunity.
153 Insight into the molecular mechanism of CIITA-
mediated inhibition of HIV-1 and HTLV
transactivators
Giovanna Tosi, 1 Chiara Orlandi,1,4 Luisa Bozzo, 1 Elisabetta Pilotti,2

Claudio Casoli,3 and Roberto S. Accolla1
1
Dept. of Clinical and Biological Sciences, University of Insubria, Varese, Italy; 2Dept. of Clinical Medicine,
Nephrology and Health Sciences, University of Parma, Parma, Italy; 3Dept. of Clinical Sciences L. Sacco,
Infectious Diseases and Immunopathology Section, University of Milan, Milan, Italy; and 4PhD, Program in
Experimental Medicine and Oncology, University of Insubria, Varese, Italy
We previously showed that CIITA, the master regulator of MHC-II genes transcription, acts as
a potent inhibitor of HIV and HTLV-2 replication by suppressing the activity of their respective viral
transactivators Tat and Tax-2. Here we show that CIITA inhibits also the activity of Tax-1 the
transcriptional activator of HTLV-1, the aetiological agent of Adult T cell Leukemia (ATL). Interestingly,
both Tax-2 and Tax-1 are inhibited by the same region of CIITA that we have now restricted to 60 aa at
the N-term of the molecule. This region is distinct from the one inhibiting the HIV-1 Tat transactivator.
Thus, our hypothesis is that Tax-1 and Tax-2 transactivators are inhibited through a common molecular
mechanism which is different from the one inhibiting HIV-1 replication. Our previous results have
shown that another transcription factor, NF-Y, which cooperates with CIITA in the activation of MHC-II
genes transcription, inhibits Tax-2 activity when over expressed in cells. A similar effect is observed also
with Tax-1 and we are assessing whether NF-Y is involved in CIITA-mediated inhibition of Tax-1 and Tax-
2. We show that Tax-1 interacts with NF-YB subunit and report on a new interaction between CIITA and
both Tax-1 and Tax-2 in vivo. Studies are also in progress to determine whether the suppressive effect
of CIITA on Tax-1 correlates with the inhibition of HLTV-1 replication as well.
These findings reveal that CIITA, beside its well known role in adaptive immunity has an important
function in innate immunity, counteracting retrovirus replication and spreading.
154 Apobec3: A Modern Twist to A Classic Retroviral
Mystery
Mario L. Santiago,1 Mauricio Montano,1 Robert Benitez,1 Ronald J. Messer,3

Wes Yonemoto,1 Bruce Chesebro,3 Kim J. Hasenkrug,3 and Warner C. Greene1,2
1
Gladstone Institute of Virology and Immunology, San Francisco, CA 94158; 2Department of Medicine,
University of California, San Francisco, CA 94143-1230; and 3Laboratory of Persistent Viral Diseases,
Rocky Mountain Laboratories, NIAID, Hamilton, MT 59840
Recovery from Friend Virus 3 (Rfv3) corresponds to a single autosomal gene encoding a resistance
trait that promotes the survival of mice infected with the Friend Virus complex (FV). Rfv3 acts by
promoting the production anti-FV neutralizing antibodies and by controlling FV viremia. Rfv3 was
discovered 30 years ago but its genetic identity has remained unknown. We now demonstrate that Rfv3
is encoded by the Apobec3 gene. Apobec3 maps to the same region of chromosome 15 as Rfv3. Of note,
the Apobec3 family of deoxycytidine deaminases are known to exert inhibitory activity against many
retroviruses including HIV. Genetic inactivation of Apobec3 converts Rfv3-resistant mice to a susceptible
phenotype characterized by diminished neutralizing antibody titers and increased viremia. Furthermore,
while Rfv3 susceptible mice contain an Apobec3 gene and produce Apobec3 mRNA, we find that
Apobec3 is naturally disabled in Rfv3-susceptible mice by aberrant mRNA splicing resulting in the
removal of exon 2 sequences. This newly discovered link between Apobec3 and neutralizing antibody
responses highlights an Apobec3-dependent mechanism of host protection that might extend to HIV.
We speculate that the ability of HIV Vif to induce degradation of Apobec3G and 3F during acute infection
could contribute to the poor neutralizing antibody response observed HIV infection.
155 Tetherin, an interferon-alpha induced inhibitor of
retrovirus release that is antagonized by Vpu
Paul D. Bieniasz
Aaron Diamond AIDS Research Center and Laboratory of Retrovirology, The Rockefeller University, New
York, NY 10016
The HIV-1 Vpu protein overcomes the action of an interferon-regulated antiviral restriction factor,
termed tetherin, which causes the retention of virions on infected cell surfaces. Using comparative gene
expression analysis and deductive constraints, we identified tetherin as a membrane protein with an
unusual topology consisting of a N-terminal cytoplasmic tail, a transmembrane domain, an ectodomain
predicted to form a coiled-coil, and anchored at the C-terminus by a putative GPI-linkage. Tetherin
expression is necessary and sufficient to restrict the release of Vpu-defective HIV-1 from human cells.
Tetherin induces dramatic accumulations of mature HIV-1 virions on the cell surfaces and, following
endocytosis, in late endosomes.
To understand mechanisms by which tetherin might function, we examined its ability to inhibit the
release of a variety of virions or virus like particles (VLPs) derived from retroviruses that share little
sequence homology. Notably, tetherin appears capable of restricting the release of every retrovirus that
we have tested. Additionally, tetherin inhibited the release of VLPs assembled by expression of the
Ebola virus matrix protein, suggesting that its activity against enveloped viral particles might be very
broad indeed, and may simply involve crosslinking of lipid bilayers. Immunofluorescence and electron
microscopy studies indicate that tetherin localizes to nascent tethered virions, consistent with the
notion that it is a physical component of the tethers that retain virions on cell surfaces.
We have cloned orthologs of tetherin from rhesus macaque, African green monkey and mouse.
These non-human tetherins are inhibit HIV-1 release, but in contrast to the human protein, are not
antagonized by Vpu. Determinants of Vpu sensitivity in tetherin reside in the transmembrane domain
and these findings suggest that HIV-1 Vpu has specifically evolved to antagonize the human tetherin
protein. Notably, Vpu only modestly affects tetherin expression levels, but abolishes tetherin’s to
localize with nascent virions, suggesting that Vpu functions by sequestering tetherin from sites of virus
particle assembly.
156 Humanized Mouse Models for The In Vivo Analysis of
HIV Infection
J. Victor Garcia-Martinez
University of Texas Southwestern Medical Center, at Dallas
HIV is predominantly transmitted by unprotected sexual contact. Currently, women worldwide

account for more than half of the estimated 11,000 newly acquired infections every day with a majority
of those transmissions occurring via the vaginal route. However, of the estimated 341,524 male adults
and adolescents living with HIV/AIDS in the US, 61% had been exposed through male-to-male sexual
contact. If untreated, the outcome of HIV infection is a systemic depletion of CD4+ T cells leading to
immunodeficiency, opportunistic infection or neoplasm resulting almost invariably in death. ART
blunts virus replication resulting in dramatic decreases in the levels of plasma viremia to undetectable
levels by standard clinical assays. This drop in plasma viremia is accompanied by increases in the level
of peripheral CD4+ T cells that remains relatively constant for the duration of ART. However, despite
this apparent disappearance of virus from peripheral blood, infected cells remain and therapy
interruption usually results in viral rebound to levels that are similar to those observed pre-treatment.
In order to advance our overall understanding of HIV and AIDS we developed and implemented
a novel small animal model where human stem cells are used to reconstitute the hematopoietic
system of immunodeficient mice. In these humanized mice (designated BLT to represent the fact they
are generated from a bone marrow transplant of mice previously implanted with a piece of autologous
human fetal liver and thymic tissue) there is systemic reconstitution with human lymphoid cells in all
hematopoietic and non-hematopoietic tissues tested. These humanized mice are susceptible to
infection with HIV-1 administered intravenously, intrarectally or intravaginally. Infection results in
plasma antigenimia, development of anti-HIV specific human antibodies, progressive depletion of
human CD4+ cells from the peripheral blood as well as systemic CD4 depletion especially in the gut
associated lymphoid tissue (GALT). This model presents unique opportunities to evaluate novel
therapeutic interventions, to advance our understanding of HIV pathogenesis and to evaluate novel
approaches aimed at virus eradication.
157 Vpr as a mediator of proteasomal degradation and
other functions
Carlos de Noronha
Albany Medical College MC151/MS216, 47 New Scotland Avenue, Albany, New York 12208
Vpr is a 15 kDa HIV1 accessory protein that promotes infection of terminally differentiated
macrophages and blocks the cell cycle of dividing cells in the G2 phase. Interestingly, HIV2 and
SIVmac/sm encode two Vpr-like proteins, Vpx and Vpr that perform these functions separately. Our
goal is to discover the mechanisms of Vpr action and their role in HIV pathogenesis by identifying the
cellular protein partners of Vpr.
Work from our lab and others revealed that Vpr from HIV1 or 2 blocks cell cycle progression by
engaging a cellular ubiquitin ligase complex. The complex, characterized by inclusion of the proteins
DCAF1, DDB1 and cullin4A must be functional for Vpr-triggered G2 arrest. We hypothesize that
a protein required for cell cycle progression is recruited to the complex by Vpr, ubiquitinated, and
thereby blocked, redistributed or degraded. Subsequent work from other labs demonstrated that both
SIV mac/sm and HIV2 Vpx also act through the same ubiquitin ligase complex to overcome an innate
block to macrophage infection that interferes with reverse transcription.
Our work to identify targets of Vpr-mediated ubiquitination revealed that HIV1 Vpr and HIV2 Vpx
both engage rpS3, a protein that is a ribosomal component, an endonuclease and a subunit of NF-kB
complexes that act on specific promoters. Previous work showing that Vpr can both stimulate and
block NF-kB action lead us to hypothesize that Vpr modulates specific NF-kB function through its
interaction with rpS3. We found that expression of HIV1 Vpr and HIV2 Vpx but not HIV2 Vpr inhibits NF-
kB reporter activity in response to TNF-a and reconfirmed that siRNA-mediated rpS3 depletion has
a similar effect. We are currently working to determine whether Vpr blocks NF-kB function by
modulating free rpS3 levels, whether the block is specific to the subset of genes that are regulated by
the rpS3-containing
NF-kB complexes and whether activity of the DCAF1/DDB1/Cullin4A ubiquitin ligase complex is
required for this Vpr function.
158 The SET complex acts as a barrier to autointegration
of HIV-1
Nan Yan,1 Peter Cherepanov,2 Janet E. Daigle,3 Alan,3 and Judy Lieberman1
1
Immune Disease Institute; 2Dana Farber Cancer Institute, Harvard Medical School; and 3Division of
Medicine, Imperial College London
Retroviruses and retrotransposons are vulnerable to a suicidal pathway known as autointegration,

which occurs when the 3’-ends of the reverse transcript are activated by integrase and then attack sites
within the viral DNA. Retroelements have diverse strategies for suppressing autointegration, but how
HIV-1 protects itself from autointegration is not well understood. We found that the SET complex, an
endoplasmic reticulum-associated complex that contains the base excision repair (BER) endonuclease
APE1 and is mobilized to the nucleus in response to oxidative stress, acts as a barrier to
autointegration. Antibodies to SET complex proteins capture HIV-1 DNA in the cytoplasm. Knocking
down any of the SET complex proteins, as well as upstream and downstream BER enzymes, increases
autointegration and decreases chromosomal integration. The SET complex likely acts to inhibit
autointegration via its presumed role in BER since enhancing uracil misincorporation by culturing cells
in the presence of added dUTP (or BrdU) or by infection with viruses encapsulating the cytidine
deaminase APOBEC3G decreases autointegration in a SET complex-dependent manner.
159 Finding Host Proteins Required for HIV Replication
Abraham Brass
Massachusetts General Hospital
HIV-1 exploits multiple host proteins during infection. Each host protein that HIV relies on is
a potential viral weakness, that once discovered represents a therapeutic target, both for prophylaxis
and treatment. We performed a large-scale siRNA screen to identify host factors required by HIV, and
identified more than 200 new, and 38 previously known, HIV-dependency factors (HDFs). These
proteins participate in a broad array of cellular functions and implicate new pathways in the viral
lifecycle. Further analysis revealed previously unknown roles for retrograde Golgi transport proteins
(Rab6 and Vps53) in viral entry, a karyopherin (TNPO3) in viral integration, and the Mediator complex
(Med28) in viral transcription. This effort begins to illustrate the power with which RNA interference
and forward genetics can be used to expose the dependencies of human pathogens such as HIV, and
in so doing identify potential targets for therapy.
160 Polarization of M1 and M2 macrophages and HIV
infection
Guido Poli
DIBIT, Via Olgettina n. 58, 20132, Milano, Italy
Permissiveness of macrophages to productive HIV-1 infection is modulated by various cytokines.

Here, we demonstrate that stimulation of primary human monocyte-derived macrophages (MDM)
with pro- (M1) and anti- (M2) inflammatory cytokines, i.e. TNF-a plus IFN-g vs. IL-4, generates two
clearly distinct populations of macrophages that differ in their ability to support CCR5-dependent (R5)
productive HIV-1 infection. Viral suppression was more profound but less sustained in M1- compared
to M2-MDM. M1 polarization was associated with a marked down-regulation of CD4, increased
secretion of CCR5-binding chemokines (i.e. MIP-a/CCL3, MIP-1a/CCL4, RANTES/CCL5) and .90%
decrease in HIV-1 DNA accumulation, suggesting inhibition of virion entry. Despite reduced virus
production in M2-MDM, CD4 was only moderately downregulated, there was no upregulation of
CCR5-binding chemokines and the early HIV-1 DNA levels were unaffected in respect to control cells.
However, M2-MDM expressed very high levels of MDC/CCL22 that we have previously linked to
a predominantly post-entry inhibition of HIV replication in MDM (M. Cota et al., PNAS 97: 9162, 2000).
Polarization also resulted in a delayed, reciprocal down-regulation of the release of M1-related
chemokines and cytokines in M2-MDM and vice versa. Most changes were fully reversible 7 days after
removal of the polarizing stimulus with the exception of IP-10/CXCL10 and MDC/CCL22, which
remained elevated in M1- and M2-MDM, respectively. In both M1- and M2-MDM, reversion to a non-
polarized state was associated with a renewed capacity to support productive infection, suggesting
a mechanism for which infected macrophage may cycle between latent and productive infection.
161 Peptide stabilization of gp120 conformation: a novel
vaccine candidate
Jonathan M. Gershoni, Anna Roitburd-Berman, and Gal Dela

Department of Cell Research and Immunology, Tel Aviv University – Israel
Binding of CD4 locks gp120 into a unique conformation, enabling chemokine-receptor

recognition necessary for viral entry. Stabilized gp120 in the CD4 induced conformation (CD4i) has
been generated via a single chain CD4-gp120 recombinant molecule which has been demonstrated to
be a preferred vaccine modality in macaques (DeVico et al 2007 PNAS 104:17477). These results
indicate that CD4-gp120 complex reveals epitopes able to elicit neutralizing antibodies; however, this
probably is at the expense of occlusion of the CD4 binding-site (BS) epitopes on gp120 - which are also
regarded as highly desirable vaccine targets. We have isolated peptides that bind gp120 and in doing
so induce the CD4i conformation - as demonstrated by binding of defining mAbs such as 17b, 19e and
CG10. Moreover, the peptide-gp120 complex continues to bind CD4, illustrating that this site remains
accessible. Furthermore, the peptide-gp120 complex efficiently binds a panel of CD4-BS defining
mAbs, such as b12, b6 and M14 thus illustrating that the CD4i conformation is compatible with
simultaneous presentation of the CD4-BS epitopes. We therefore propose that the peptide-gp120
complex represents a novel vaccine candidate that benefits from ‘‘both worlds’’; a CD4i stabilized
conformation that maintains accessible neutralizing epitopes associated with the CD4-BS of gp120.
162 Transmission Networks of Drug Resistance Acquired
in Primary/Early Stage HIV Infection
Brenner BG, Roger M, Moisi D, Oliveira M, Hardy I, Turgel R, Charest H,

Routy J-P, and Wainberg MA, the Montreal PHI Cohort HIV Prevention Study
Groups
Jewish General Hospital – McGill University AIDS Centre, 3755 Cote Ste-Catherine Road, Montreal,
Quebec H3T 1E2
Objectives: Population-based sequencing of primary/recent HIV infections (PHI) can provide

a framework for understanding transmission dynamics of local epidemics. In Quebec, half of PHI
represent clustered transmission events. This study ascertained the cumulative implications of
clustering on onward transmission of drug resistance.
Methods: HIV-1 pol sequence datasets were available for all genotyped PHI (,6 months post-
seroconversion (n = 848 subtype B infections, 1997–2007). Phylogenetic analysis established clustered
transmission events, based on maximum likelihood topologies having high bootstrap values (.98%)
and short genetic distances. The distributions of resistance to nucleoside and non-nucleoside RT
inhibitors (NRTIs and NNRTIs) and protease inhibitors (PIs) in unique and clustered transmissions
were ascertained.
Results: Episodic clustering was observed in half of recent/early stage infections from 1997–2008.
Overall, 29% and 28% of new infections segregated into small (,5 PHI/cluster, n = 242/848) and large
transmission chains ($5 PHI/cluster, n = 239/848), averaging 2.8 6 0.1 PHI/cluster and 10.3 6 1.0
PHI/cluster, respectively. The transmission of nucleoside analogue mutations and 215 resistant variants
(T215C/D/I/F/N/S/Y) declined with clustering (7.9% vs. 3.4% vs. 1.2% and 5.8% vs. 1.7% vs. 1.1% for unique,
small and large clustered transmissions, respectively). In contrast, clustering was associated with the
increased transmission of viruses harbouring resistance to NNRTIs (6.6% vs. 6.0% vs. 15.5%, respectively).
Conclusions: Clustering in early/PHI stage infection differentially affects transmission of drug
resistance to different drug classes. Public health, prevention and diagnostic strategies, targeting PHI,
afford a unique opportunity to curb the spread of transmitted drug resistance.
163 The State of the ART in HIV Therapeutics
John G. Bartlett
1830 East Monument St., Rm. 447, Baltimore, MD
Presentation based on guidelines from U.S. and Europe (which are nearly identical).
When to start:
2 NRTIs: TDF/FTC, ABC/3TC, ?AZT/3TC
3d drug: EFV, LPV/r, ATV/r, FPV/r, SQV/r
When to change:
Toxicity, convenience or virologic failure (defined as VL. 50 c/mL 3 2)
What to change to:
Toxcitiy - single drug substitution
Viral failure - based on resistance testing
Major issues:
1. Should HIV screening be ‘‘routine’’ for all?
2. Should everyone be treated except long-term progressors?
3. Does ART play an important role in prevention?
4. When will point-of-care diagnostics be available in clinics for patients?
164 An update on raltegravir (Isentress)
Charles Farthing, MD
U.S. Dir of Scientific Affairs for Merck & Co Inc., 770 Sumneytown Pike, WP97A-279, PO Box 4, West Point, PA
19486-0004
Raltegravir, the first in class integrase inhibitor from Merck, will be presented with an update of
phase 2 and 3 clinical trials, and an update on PK, drug – drug interaction data, and resistance data,
obtained to date.
96 week phase two data in na¨ve patients shows a sustained virological suppression of viral load
with TNF+FTC+RAL similar to the control arm of TNF+FTC+EFV with all four doses of RAL studied – 100
mg, 200 mg, 400 mg and 600 mg bid. Drug related clinical adverse events with RAL at 48%, were less
then with EFV - 71%. RAL had a neutral effect on serum lipids.
Suppression of viral load with RAL was somewhat more rapid initially with RAL than with EFV
although the relevance of this finding is unknown.
48 week data in two phase three studies with RAL and OBR (BENCHMRK) show sustained
virological suppression to ,50 copies/mL in 62% of patients versus 33% with OBR alone. Side effects
for patients treated with OBR+RAL were similar as for those treated with OBR alone suggesting RAL
added little to toxicity.
PK with RAL is highly variable and no relationship between blood levels and results has so far
been determined. RAL is metabolized by glucuronidation by the enzyme UGT1A1. Studies to date
show no significant drug level alterations with different UGT1A1 polymorphisms - and few concerning
drug drug interaction with drugs that induce or inhibit UGT1A1. Tipranavir lowers RAL levels by 55%
and rifampin lowers levels by 61%. Phase 3 data suggests tipranavir’s effect is not significant – no data
yet exists for use with rifampin.
Resistance to RAL usually follows one of two pathways with primary mutations at either position
Q148 or N155, which are usually rapidly followed by the development of secondary mutations which
increase resistance and return fitness to the virus.
165 Evaluation of Efficacy and Immune Recovery of
Optimized Background Therapy (OBT) Plus
Maraviroc (MVC) vs Placebo (PBO) in Treatment
Experienced (TE) Patients with only R5 HIV-1,
Combined Analysis of MOTIVATE 1 and 2
R. Tressler,1 J. Goodrich,2 N. Rajicic,1 H. Valdez,1 and H. Mayer2

1
Pfizer, Inc., New York; 2Pfizer, Inc., New London
Intro: In studies with MVC a numeric increase in CD4 cell count has been observed in the groups
receiving MVC vs Placebo or efavirenz. The clinical parameters associated with and clinical significance
of the increase in CD4 cell count are unknown. This post hoc analysis of the Wk 48 data combined
MOTIVATE 1 and 2 data explores efficacy of MVC by degree of immune deficiency at BL, the clinical
relevance of and predictors of the CD4 cell count increase observed in the MVC vs PBO arms.
Methods: Descriptive statistics of antiviral efficacy (ITT), time to virologic failure, change in CD4
cell count (LOCF) and time to a Category C event were computed for all patients who received at least
one dose of randomized study drug. A multivariate analysis of BL demographic and disease
characteristics was conducted to determine parameters of changes in CD4 count at Wk 48.
Results: There were no differences at BL among randomized treatment arms in CD4 cell count,
viral load, age, gender, race and number of new drugs used in OBT. At all clinically relevant CD4 cell
count strata a greater proportion of patients receiving MVC achieved HIV RNA ,50 copies/mL at Wk 48
as compared to the PBO arm.
Conclusions: Efficacy of MVC was observed in all CD4 strata and CD4 cell increases were greater in
patients who received MVC vs PBO independent of viral load change or achieving a viral load ,50
copies/mL. Predictors of CD4 count increase, other than MVC use, were similar to what has previously been
reported. The higher proportion of patients achieving CD4 cell count .200 cells/mm3 on MVC, and the fact
that on treatment CD4+ cells (but not treatment arm) were associated with lower risk of Category C events,
suggests a clinical relevance to the CD4 cell count increases on MVC observed in the MOTIVATE studies.
(Continued on next page)
165 Evaluation of Efficacy and Immune Recovery of
Optimized Background Therapy (OBT) Plus
Maraviroc (MVC) vs Placebo (PBO) in Treatment
Experienced (TE) Patients with only R5 HIV-ombined
Analysis of MOTIVATE 1 and 2
(continued)
Placebo (n = 209*) MVC QD (n = 414*) MVC BID (n = 426)

HIV RNA ,400 copies/mL 22.5% 51.7% 56.1%
HIV RNA ,50 copies/mL 16.7% 43.2% 45.5%
Median change in CD4 cell count at +24 +92 +103
Wk 48 (cells/mm3)
Median change in CD4 cell count at Wk 48 +96 (n = 68) +125 (n = 251) +126 (n = 267)
for pts who achieved HIV RNA ,50 copies/mL
at least once (cells/mm3)
Median change in CD4 cell count at Wk 48 for pt +10 (n = 140) +43 (n = 162) +59 (n = 159)
who never achieved HIV RNA ,50 copies/mL (cells/mm3)
Percent of patients with CD4 ,200 at BL who 32 (n = 118) 43 (n = 235) 47 (n = 250)
achieved .200 at Wk 48
Percent of patients who experienced a Category C event 7.7% 7.0% 5.2%
through Wk 48 (unadjusted for exposure)
*One patient in the PBO and MVC QD arm were missing BL CD4 data, therefore CD4 cell counts are based on 208 pts in the PBO and 413 pts in the MVC QD groups. (Time to
occurrence of a Category C event was statistically significantly longer for MVC (QD or BID) vs PBO arms. Higher on-treatment CD4 cell counts were independently associated with
a decreased risk of a Category C event. The discontinuation rate and adverse event profile was similar in the MVC and PBO arms. BL CD4 cell count, BL viral load, change in viral load at
Wk 48, age and MVC use were independent predictors of CD4 cell increases.
166 HIV Therapeutics in Resource Limited Settings –
Status and Future Directions
Anthony Amoroso
Institute of Human Virology, 725 West Lombard Street, Baltimore, MD 21201
In 2003 there was an estimated 50,000 patients on antiretroviral therapy in sub-Sahara Africa.
Today there is an estimated 2 million. Concerns regarding efficacy and durability of antiretroviral
therapy remain. In addition the potential for race/gender specific drug toxicity profiles in the targeted
populations exist. This session will review data on long term durability of first line regimens used in
large scale-up treatment programs to include toxicity and efficacy.
167 Update on Gilead Sciences Anti-HIV Development
Programs
Tomas Cihlar, PhD

Gilead Sciences, Inc., 333 Lakeside Drive, Foster City, CA 94404
Gilead Sciences has two current development programs for anti-HIV small molecule inhibitors.
Elvitegravir (EVG, GS-9137) is an HIV-1 integrase inhibitor (INI) which has completed a phase 2 dose-
ranging study (Study GS-US-183-0105). Patients were randomized to 20 mg, 50 mg or 125 mg QD
doses of ritonavir-boosted EVG (EVG/r) plus optimized background therapy (OBT) comprised of
NRTIs +/2 T-20. Study 0105 patients were highly experienced with a median of 3 TAMs and 11 PI-R
mutations at baseline. Patients receiving EVG 125 mg had rapid viral load declines (mean decline in
HIV-1 RNA of 2.1 log10 copies/mL at week 24); those with $1 active drug(s) in their background
therapy had durable responses. Among patients with virologic failure on EVG/r 125 mg, the most
common integrase mutations detected were E92Q, E138K, Q148R/K/H and N155H (observed in 11/28,
39%), S147G (9/28, 32%) and T66I/A/K (5/28, 18%), some of which have been selected by raltegravir
(MK-0518) in vivo. RC declined from a median of 108% at baseline to 54% at virologic failure. EVG is
currently entering phase 3 studies.
In another program, a novel nucleotide analog, phosphonomethoxy-2’-fluoro-2’, 3’-dideoxydi-
dehydroadenosine (GS-9148), was selected as an NRTI with an improved pharmacological and
resistance profile. The prodrug of GS-9148 (GS-9131) exhibits potent antiretroviral activity both in
primary cells and T-cell lines (EC50 = 25-200 nM), low cytotoxicity (CC50 . 50 iM) in multiple cell types,
and shows no effect on mitochondrial DNA or lactate production in HepG2 cells. In phenotypic assays,
the activity of GS-9148 was not affected by the K65R, L74V, or M184V mutations in RT (EC50 fold
change ,1). Viruses with 4 or more thymidine analog mutations showed 0.74 to 2.0-fold change in the
susceptibility, a shift that was smaller than that of any other tested NRTI. Passage of HIV-1 in the
presence of GS-9148 selected for a primary K70E mutation and two other RT mutations that together
conferred ,3-fold resistance to GS-9148. Oral administration of GS-9131 in dogs resulted in high and
persistent levels of GS-9148 diphosphate in PBMCs and lymph nodes. Overall, GS-9131 exhibits
a favorable in vitro virological and in vivo pharmacological profile and is entering phase I trials for once
daily dosing in NRTI-experienced patients.
168 Rapamycin enhances the antiviral activity of CCR5
antagonist Vicriviroc
Olga Latinovic
Institute of Human Virology School of Medicine, University of Maryland, 725 West Lombard Street,
Baltimore, MD 21201
The CCR5 chemokine receptor plays a crucial role in human immunodeficiency virus type I (HIV-1)
infection. We previously demonstrated that reduction of CCR5 density with low doses of the transplant
drug Rapamycin (RAPA) potentiates the antiviral activity of HIV-1 fusion inhibitor Enfuvirtide (T-20)
against R5 strains of HIV-1 [Heredia et al, AAC, 2007]. In the current study, we show that RAPA enhances
the potent antiviral activity of the CCR5 antagonist, Vicriviroc (VCV), which is currently being evaluated
in Phase III clinical trials.
The goal of this investigation is to identify most effective combination of antiviral drugs with the
lowest level of side effects. Drug interaction based studies show that the RAPA/VCV combination has
significant antiviral synergy (combination indexes of 0.1–0.04) in both spreading-cycle and single-cycle
infection of lymphocytes, as well as in a cell-cell fusion assay. The drug, RAPA, enhances VCV antiviral
activity against both B and non-B clade isolates, potently suppressing clade G viruses with reduced
sensitivities to VCV and to Maraviroc, a second CCR5 antagonist. An existing synergy between RAPA
and VCV translated into Dose Reduction Indices (DRI) results in 8–41 fold reduction for RAPA and 19–
658 fold reduction for VCV. In addition, we found that the RAPA induced reduction of CCR5 density in
lymphocytes increased sensitivity to VCV in VCV-resistant strains, inhibiting virus production by ;90%.
Moreover, by using cell lines and donor lymphocytes expressing a range of CCR5 densities, we further
demonstrate the importance of CCR5 density to VCV activity. These research results direct to the future
avenue of enhancing the potency of CCR5 antagonists, reducing potential for toxicity, potentially
controlling emerging VCV-resistant variants.
169 Why do so few Antibodies Neutralize HIV-1? Tests of
the ‘‘Tolerance Hypothesis’’
Garnett Kelsoe
Department of Immunology, Duke University, Durham, North Carolina 27710 USA
The emergence of serum antibody responses only after early cellular immune responses have
suppressed HIV replication has led to doubts regarding the importance of humoral immunity in
controlling HIV infections. Nonetheless, rare, broadly cross-reactive antibodies are capable of
neutralizing multiple isolates of HIV-1 in vitro, and when passively administered to monkeys, prevent
experimental infection by SHIV. Why are such antibodies rarely produced by HIV-infected patients?
Recently, several of these rare, HIV neutralizing antibodies were shown to react with self-antigens
leading to the possibility that effective HIV humoral responses are constrained by the tolerization of
HIV-reactive B cells that also recognize self-antigens. We have begun to test the hypothesis that B cells
that recognize phylogenetically conserved, neutralizing epitopes of the HIV-1 gp-41 membrane
proximal external region (MPER) are present in early, developmentally immature B cells but are
tolerized and lost during their subsequent development/maturation. Our studies indicate that murine
B cells specific for the 2F5 and 4E10 MPER epitopes are present at significant frequencies in the
immature and transitional B-cell compartments of bone marrow but are undetectable in peripheral B-
cell pools including phenotypically identical transitional B cells. Perhaps for this reason, immunization
of normal mice with MPER peptides containing the 2F5 and 4E10 epitopes peptides results in little
antibody production and poor germinal center responses. In contrast, reconstitution of RAG1 deficient
mice with B lymphocytes that develop outside of the tolerizing environment of the bone marrow is
permissive both for robust autoantibody responses, the production of germinal centers, and greatly
increased levels of IgG antibody to MPER peptide epitopes.
170 Getting the Right Immune Response to HIV:
Evaluation of Protective Immune Responses After
Vaccination of Rhesus Macaques
George N. Pavlakis,1 Antonio Valentin,1 Margherita Rosati,1 Agneta von Gegerfelt,1

Vainav Patel,1 Cristina Bergamaschi,1 Brunda Ganeru,1 Osamu Usami,1
Viraj Kulkarni,2 Rashmi Jalah,2 Candido Alicea,2 and Barbara K. Felber2
1
Human Retrovirus Section; and 2Human Retrovirus Pathogenesis Section, NCI, Frederick
Rhesus macaques infected by SIV develop a disease remarkably similar to human AIDS and
provide the best animal model for AIDS vaccine evaluation. We have used this model to study immune
responses after vaccination and challenge with pathogenic heterologous stocks of SIV. Our experience
includes DNA vaccination either alone or in combination with recombinant virus or protein boost, and
also vaccination with attenuated non-pathogenic (or weakly pathogenic) SIV molecular clones.
Analysis of immune responses from several vaccination/challenge experiments show the development
of protective immune responses that prevent high viremia in the challenged macaques. The correlates
of protection remain poorly defined. It is anticipated that more detailed analysis of immune responses
will provide a better understanding of the nature of protective immune response.
The methodology of DNA vaccination has advanced rapidly. In the past it produced meager
results in primates and humans, whereas at present DNA vaccination with optimized vectors and
delivery is shown to efficiently produce high and durable cellular and humoral immune responses.
These responses are disseminated to mucosa and protect the animals from high viremia after
challenge.
The best protection in the SIV macaque model has been achieved by the use of non-pathogenic,
attenuated SIV clones. Such animals are able to fully suppress the pathogenic challenge virus for many
years and protect the animals from disease development. Comparison of immune responses of
animals protected after different vaccine regimens suggest that the quality, timing and location of
immune responses are critical. Better assays to evaluate immune responses are essential.
171 Induction of Cross-clade Neutralizing Antibodies in
Rabbits Using a DNA Prime/Protein Boost
Immunization Regimen
S. Zolla-Pazner,1,2 S. Cohen,2 C. Krachmarov,3 A. Pinter,3 S. Wang,4 and S. Lu4

1
Veterans Affairs Medical Center, New York, NY 10010; 2Dept. of Pathology, NYU School of Medicine,
New York, NY 10016; 3Public Health Research Institute, Newark, NJ 07103; and 4University of
Massachusetts Medical School, Worcester, MA 01605
Previous studies showed that an immunization regimen that focuses the immune response on the
structurally conserved V3 epitope of gp120 can result in the induction of cross-clade neutralizing
antibodies (Abs). Experiments were undertaken to optimize the breadth and strength of this response
by immunizing rabbits with three doses of gp120 DNA containing a V3 loop characterized by the GPGR
motif at its tip, and/or with gp120 DNA with a V3 loop carrying the GPGQ motif. Priming was followed
by protein boosts of a V3-gp70 fusion protein carrying the V3 sequence from a clade B virus with the
GPGR motif (V3B-FP), a V3A-FP with a clade A V3 loop with the GPGQ motif, and/or a V3C-FP (GPGQ
motif). Neutralization was demonstrated against viruses pseudotyped with the SF162 envelope in
which V3 was replaced with consensus V3 loops from subtypes A1, CRF02_AG, B, C, CRF01_AE, F or H.
.50% neutralization was also achieved at titers ranging from 1:10–1:559 against 14 of 15 primary
isolates from the four subtypes tested (A, CRF02_AG, B and C). The polyclonal Abs in the immune
rabbit sera were able to neutralize viruses that none of 10 human anti-V3 mAbs could neutralize. Thus,
cross-clade neutralizing Abs were induced using a DNA prime/protein boost regimen designed to
focus the immune response on a neutralizing epitope, The broadest neutralizing responses were
generated when using a gp120 DNA prime derived from a clade C virus carrying a V3 loop with the
GPGQ motif.
172 Prospects for an AIDS Vaccine: Can Effector-Memory
T Cell Responses Contribute?
L. Picker
Vaccine and Gene Therapy Institute and the Oregon National Primate Research Center at Oregon
Health & Science University, Beaverton, OR
Memory T cells, both CD4+ and CD8+, can be described in terms of their differentiation along the
so-called ‘‘central-memory’’ (TCM) to ‘‘effector-memory’’ (TEM) axis. TCM recirculate among secondary
lymphoid tissues and exhibit the capacity to expand and under-go further differentiation to TEM or
active effectors upon antigenic stimulation. In contrast, TEM populations are based primarily in non-
lymphoid tissues, have less proliferative potential upon re-stimulation than TCM, but can manifest
potent immediate effector function. TEM would therefore provide an immediate, but limited, response
to microbial invasion of non-lymphoid tissues, whereas TCM would potentially provide a much larger,
but delayed, response. Prime-boost vaccines or vaccines with non-persistent vectors typically yield
TCM predominant responses that are dependent on post-challenge expansion for effective effector
generation. The limited efficacy of these approaches to date might be due to a kinetic mismatch
between this delayed production of T cell effectors and the rapid replication and diversification of
HIV/SIV after transmission. As TEM are pre-positioned and ‘‘armed’’ at sites of initial HIV/SIV replication,
vaccines that elicit and maintain these populations might have an advantage in mediating anti-HIV/SIV
protection early, in the window of opportunity prior to massive systemic viral replication. To test this
hypothesis, we have developed CMV-based vectors that are able to prime and maintain strong SIV-
specific TEM responses in rhesus macaques. In keeping with our hypothesis, these SIV-specific TEM
responses endow resistance to the acquisition of pathogenic SIV infection delivered by a mucosal
route, but do not provide substantial protection when progressive systemic infection does occur.
These data suggest that vaccines capable of generating and maintaining HIV-specific TEM might be
able to decrease the incidence of HIV infection after sexual exposure.
173 Human defensins – small in size but big in
functionality
Wei Gang, 1 Erik de Leeuw, 1 Guozhang Zou, 1 Mohsen Rajabi, 1

Marzena Pazgier,1,2 Weirong Yuan,1 Jing Li,1 Jacek Lubkowski,2 and
Wuyuan Lu1
1
Laboratory of Synthetic Protein Chemistry, Division of Basic Science and Vaccine Research, Institute of
Human Virology, University of Maryland School of Medicine, 725 West Lombard Street, Baltimore, MD
21201, USA; 2Macromolecular Assembly Structure and Cell Signaling Section, NCI, National Institutes of
Health, Frederick, MD 21702, USA
Defensins are a family of small cationic peptides expressed primarily in leukocytes and epithelial
cells. Lehrer and colleagues originally discovered, two decades ago, the proto-
type alpha-defensins – human neutrophil peptides or HNPs – as antibiotic peptides important for
phagocytosis. Recent studies have unveiled a great variety of new functions of these antimicrobial
molecules in many biological processes. However, the molecular basis for the functional promiscuity of
defensins remains poorly understood. Using an extensive mutational analysis aided by structural
biology, we have systematically investigated the structural determinants for HNP1 as a bactericidal
agent and a potent inhibitor of anthrax lethal factor. As a result, a clear picture of how human
defensins work at the molecular level is beginning to emerge.
174 The C5 region of gp120: A therapeutic vaccine
target?
Angus Dalgleish, and Martin Cadogan

SGUL, tooting, London SW17 ORE, UK
HIV infection does not always lead to disease as demonstrated by very long term non progressors
and chimpanzees. The difference between progressors and non progressors is the degree of non
specific pan activation of the immune system which is absent in the latter. The cause of this genetically
restricted pan activation has yet to be confirmed. However, recognized causes such as hypervariability
and superantigen stimulation are not relevant to HIV. Another potential cause is allogeneic stimulation
. HIV has several regions of homology to HLA class 1 and 11. We have reported that C5 is structurally
very similar in that it and no other region bind specific class 1 and class 2 peptides. Moreover, when the
region is itself presented as a peptide in context of self HLA it is recognized as an alloepitope . As such
it would be closest to a self peptide in HLA-27 and most pronounced as an alloepitope in HLA-8.
This is consistent with HLA-27 being associated with slow progression and HLA-8 with fast
progression. Furthermore, chimps have very restricted HLA types similar to HLA-27/57.
Several large studies have reported the presence of anti-bodies to C5 in non progressors but
these observations have been discounted as the antibodies are non neutralizing.
Here, we propose that if these antibodies are preventing the C5 region from inducing the
activation that drives AIDS, then a therapeutic vaccine aimed at inducing this response following
immune normalization with HAART may allow some patients to convert to non progressors and no
longer be dependant on HAART therapy.
As this region is so highly conserved amongst all viable isolates it may well be a good candidate
for a prophylactic vaccine as sterilising immunity is clearly not necessary in order to prevent
progression to AIDS.
Therapeutic non-neutralising vaccines need not be limited to HIV!
175 Mechanisms by which synergistic combinations of
TLR ligands enhance T cell responses to vaccines
Qing Zhu, 1 Igor M. Belyakov, 1 Dennis Klinman, 2 and Jay A.

Berzofsky1
1
Vaccine Branch and 2Laboratory of Experimental Immunology, Center for Cancer Research, National
Cancer Institute, NIH, Bethesda, MD 20892 USA
Ligands for Toll-Like Receptors (TLRs) are usually microbial products that are naturally
encountered in combinations. Thus, we asked whether synergies occurred among them, and what
the mechanisms of those synergies might be, and how we could exploit that for enhancing vaccine-
induced T cell responses. We found several synergies between pairs of TLR ligands that used different
intracellular signal transduction pathways. In each case, the synergy depended on dendritic cell (DC)
activation either in vitro or in vivo to induce an enhanced T cell response. Dendritic cells were similarly
activated both in vitro and in vivo, and when treated with the synergistic combinations of TLR ligands
in vitro, the DCs could be used in vivo as a vaccine to induce an enhanced immune response. The
increased responses correlated with dendritic cell production of cytokines like IL-12, that depended on
the MyD88 pathway, but not with increased costimulatory molecules, that depended only on the TRIF
pathway. However, the TRIF pathway was found to amplify the cytokine production through the
MyD88 pathway in a unidirectional crosstalk, accounting for the synergy. The synergistic combinations
activate CD4+ and CD8+ T cells independently. Taking advantage of these synergistic combinations of
TLR ligands could allow the design of more effective vaccine compositions and adjuvants and
potentially overcome deficiencies in DC maturation or lack of CD4+ help in cancer or HIV infection.
176 Vector-based Vaccines for Cancer Therapy
Jeffrey Schlom, James W. Hodge, Kwong-Yok Tsang, Claudia Palena,

John W. Greiner, Ravi A. Madan, and James L. Gulley
Laboratory of Tumor Immunology and Biology, Center for Cancer Research, NCI, NIH
Our program has been involved in the design and development of recombinant vector based
vaccines for the therapy of human carcinomas. Emphasis has been based on the development of a pox
virus-based regimen, i.e., recombinant vaccinia prime vaccination followed by multiple booster
vaccinations with recombinant fowlpox vectors. Clinical trials have demonstrated that one can
administer multiple boosts of recombinant fowlpox without host neutralizing activity. Vaccines have
now been developed which contain transgenes for three human T-cell costimulatory molecules
(TRICOM) and transgenes for either PSA for prostate cancer vaccines, or CEA-MUC-1 for use in the
therapy of most carcinomas.
These vaccines have demonstrated evidence of antigen-specific T-cell responses, the initiation of
antigen cascade, objective clinical responses, drops in serum markers, delays in time to progression,
and most importantly, evidence of increase in patient survival. In two separate clinical trials, evidence
of increased survival has been observed in prostate cancer patients with hormone refractory disease
and with metastatic disease, respectively.
Preclinical studies have clearly demonstrated that vaccine-induced T-cell immunity can act
synergistically with radiation therapy, and certain chemotherapeutic agents by altering the phenotype
of tumor cells to make them more susceptible to T-cell–mediated attack. Evidence is also emerging
that the dynamic process of T-cell–mediated immunity initiated by vaccine therapy can have a positive
influence on subsequent therapies.
REFERENCE
1. Schlom, J, P.M. Arlen, J.L. Gulley. 2007. Cancer vaccines: moving beyond current paradigms. Clin. Cancer Res. 13:3776–3782.
177 Recruitment of high avidity antigen-specific T cells
in pancreatic cancer patients
Elizabeth M. Jaffee, MD
Sidney Kimmel Cancer Center at Johns Hopkins, 4M07 CRB I, 1650 Orleans St., Baltimore, MD 21231
T cell responses are often observed in cancer patients who have been treated with antigen-
specific vaccines. However, the majority of responses are often weak and ineffective at controlling
tumor growth. In some instances, this may be due to an ineffective vaccine approach. However, in
many cases mechanisms of T cell tolerance or ‘‘immune checkpoints’’ to specific tumor antigens are at
play. Understanding these mechanisms in the context of tumor antigens is critical for the development
of interventions that can reverse the tolerant state or modify ‘‘immune checkpoints’’ and allow these T
cells to more effectively respond to tumors. We have described the existence of immune tolerance in
the HER-2/neu transgenic (neu-N) mouse model of breast cancer and used these mice to understand
the mechanisms that suppress high avidity antigen-specific CD8+ T cells. We previously reported that
treatment of neu-N mice with immune modulating doses of Cyclophosphamide (Cy) prior to
vaccination resulted in tumor protection in a proportion of mice. More recently we reported that CD8+
T cells specific for the immunodominant neu epitope, RNEU420-429, were identified only in Cy plus
vaccine treated neu-N mice that rejected tumor challenge, but not in neu-N mice given vaccine only.
Furthermore, high avidity RNEU420-429-specific CD8+ T cells were also identified in vaccine treated
mice that were first depleted of CD25+ T regulatory cells (Tregs). Thus, the mechanism by which
immune modulating doses of Cy augments vaccine induced high avidity RNEU-specific CD8+ T cell
responses is by inhibiting and/or deleting CD4+CD25+ Tregs. ‘‘Proof of principle’’ clinical trials have
confirmed that vaccines given with immune modulating Cy can recruit higher avidity T cells that
correlate with improved clinical outcomes. The antigen-specific T cell responses associated with these
clinical trials will be discussed.
178 Development of Preventive HIV Vaccines in the US
Military HIV Research Program
Nelson L. Michael, MD, PhD

Approaches to develop a preventive HIV-1 vaccine have been challenged by the failure of protein
subunit and some non-replicating vector immunizations to reduce HIV-1 acquisition risk or post-
infection viremia in breakthrough infections. Recent trials of adenovirus type 5 vectored HIV-1 vaccines
have raised concerns that some vaccines under certain circumstances may increase HIV-1 acquisition
risk. Long-standing challenges to develop vaccines that produce functional B-cell responses, effective
T-cell responses, mucosal immunity, and that work across the wide range of HIV-1 sequence variation
have not been overcome. The HIV vaccine development field has undergone much introspection
resulting in greater emphasis on vaccine discovery research, relevant small animal and non-human
primate models, and new approaches to vaccine delivery. The US Military HIV Research Program has
major efforts that address this new paradigm. Clinical data will be presented from novel MVA vectored
vaccine phase I trials and planned experiments with heterologous vector approaches will be
described. Finally, an update on the progress of the phase III trial of ALVAC-gp120 protein in Thailand
will be provided.
179 Rational Vaccine Design and the Development of an
AIDS Vaccine
Gary J. Nabel, MD, PhD

Vaccine Research Center, NIAID, National Institutes of Health, Bethesda, MD 20892-3005, USA
Advances in our basic understanding of the immune system have provided the tools to make
more effective vaccines—rationally designed vaccines developed with a scientific understanding of
the mechanisms of microbial pathogenesis and immunity. Current HIV vaccine candidates target
multiple internal and external gene products. Next generation vaccines will need to enhance the
immunogenicity of the envelope (Env), with the goal of improving the breadth of the neutralizing
antibody response while concurrently stimulating cell-mediated immunity. Our recent efforts have
centered on the use of genetic and structural biological information to improve both Env and Gag
immunogen design. The genetic approach has focused on the conserved regions of Gag and Env to
improve CD8 cell breadth. To target such conserved domains, site-specific mutagenesis has been
employed to generate and analyze mosaic, variant, and chimeric Env and Gag proteins. In other
studies, the VRC and collaborators have determined the structure of the broadly neutralizing
monoclonal antibody, b12, as well as its corresponding HIV-1 epitope. This information is being used
to design epitope mimics that can induce additional insight into the immune response. The status of
the rational immunogen design and the development of vaccines that elicit broadly neutralizing
antibody responses will be reviewed.
180 Continuing Clinical Trials for HIV Vaccine Research
Glenda Gray
PHRU, Chris Hani Baragwanath Hospital, Old Potch Road, Diepkloof, Soweto, Johannesburg
There is a pressing need to continue developing novel biological platforms that will interrupt HIV
transmission as is evident from the explosive epidemic seen in South Africa. South Africa, with an
estimated 5.7 million infected with HIV, where 90% of all new infections amongst 15–24 year olds
occur in girls and women, is in urgent need for a biomedical intervention that is not coital dependent.
HIV Vaccine related research presents many challenges to scientists in the post ‘‘STEP’’ era. These
challenges include the continued need for discovery, design and manufacturing of candidate vaccines
as well designing clinical trials that address specific immunological and virological questions that
advance our understanding of HIV and impact on immunogen design and development. Continuing
clinical research that addresses genetic diversity of HIV-1, host immune responses to HIV-1, anti-vector
immunity, and genetic susceptibility to HIV-1 are critical. The nature of protective CTL remains
unidentified and can only be revealed by doing further clinical testing. The successful execution of this
will require mechanisms for continued global financing and commitment to basic science clinical trials.
181 Gunnel Biberfeld
Abstracts not available at time of printing
182 A Novel Model for In Vivo SIV Neutralization
Phil Johnson
Children’s Hospital of Philadelphia, 1216B ARC, 3615 Civic Center Blvd, Philadelphia, PA 19104
The holy grail of HIV vaccine development is an immunogen that elicits antibodies which broadly
neutralize field strains of the virus. While tremendous insights have been gained into the structure and
function of the HIV envelope glycoprotein, precious little progress has been made in designing such
immunogens. In fact, it remains formally possible that with current technologies, engagement of the
adaptive immune system will not lead to an effective HIV vaccine. Considering this possibility, we have
taken a markedly different approach to the generation of serum neutralizing activity against HIV that
we have dubbed ‘‘reverse immunization.’’ In this model, we pre-select the antibody or antibody-like
molecule of interest (i.e, one that broadly neutralizes) then synthetically derive the representative gene
and place it into the context of an adeno-associated virus (AAV) gene transfer vector. When injected
intramuscularly into a vaccinee, the AAV vector genome directs in vivo production of the gene product
that leads to serum neutralizing activity against HIV. Recently, we have refined this approach for the
SIV model system, and can now provide in a single dose, sterilizing immunity to vaccinated monkeys
against a virulent SIV challenge virus. Thus, reverse immunization holds significant promise as
a ‘‘designer’’ approach to an effective HIV vaccine.
183 Morphogenomic immune responsiveness to
preventive/therapeutic vaccines
A. Monaco,1 M. Sabatino,1 Z. Pos,1 D. F. Stroncek,1 E. Wang,1 R. C. Gallo,2

G. K. Lewis,2 M. L. Tornesello,3 F. M. Buonaguro,3 F. M. Marincola,1 and
L. Buonaguro3
1
Immunogenetics Section, Dept Transf Medicine, Clinical Center, NIH, Bethesda; 2Inst. Human Virology,
Univ. of Maryland School of Medicine, Baltimore, MD, USA; 3Lab. of Viral Oncogenesis and
Immunotherapies & AIDS Reference Center, Istituto Nazionale Tumori ‘‘Fond. G. Pascale’’, Naples – Italy
We have previously reported that the transcriptional profile of VLPstimulated MDDCs shows the
up-regulation of classic markers of MDDC activation. In particular, VLP-stimulation induced the
activation of genes associated with antigen presentation and MDDCs migration. Subsequently, we
have shown that a comparable transcriptional profiling can be obtained using PBMCs without further
in vitro maturation in MDDCs. Fresh human peripheral blood mononuclear cells were isolated by
Ficoll-Hypaque density gradient centrifugation and plated in 6-well plates at a concentration of
1 3 107/well. PBMCs were pulsed with 6 mg/mL of HIV-VLPs. After 16 hrs, the cells were harvested,
washed, total RNA was isolated and RNA quality/quantity was estimated. Amplified antisense RNA
(aRNA) was obtained from total RNA via two round of in vitro transcription. Testreference sample pairs
were mixed and co-hybridized to a custom-made 37K oligo-based microarray platform encompassing
the whole human genome. In the present study we show that VLPs induce ex vivo transcriptional
activation comparable to that observed in in vitro matured MDDCs and the transcriptional patterns
induced ex vivo by VLPs in PBMCs from normal donors equated the patterns observed ex vivo in
unstimulated PBMC from HIV in patients. Moreover the baseline activation of immune genes observed
in HIV-infected patients is further enhanced by ex vivo stimulation by VLPs resulting in a transcriptional
activation superior to that observed in PBMC obtained from normal non HIV-infected volunteers. In
summary, this study may provide a road map for the study of HIV-infected patients based on
HIVspecific signatures, their heterogeneity among patients, their response to vaccine strategies. Future
studies should take into account this findings for the discovery of potentially correlative patterns
associated with the natural history of the disease and/or it response to treatment.
184 Development of the broadly neutralizing human
antibody m9 for anti-HIV prophylactics
Antony S. Dimitrov, Wenlei Zhang, Ilia Prado, and Timothy Fouts

Profectus BioSciences, Inc., 1450 South Rolling Road, Baltimore, MD 21227
ScFv m9 is a human antibody fragment that exhibits potent and broad anti-HIV activity. Identified
by using phage display, scFv-m9 targets a highly conserved CD4-induced (CD4i) epitope on gp120 and
potently neutralizes Groups M (clades A-G) and N HIV-1 in PBMC/primary isolate-based and cell
line/pseudovirus-based assays most likely by interfering with envelope engagement to the coreceptor.
This monoclonal antibody fragment neutralized more than 80% of the tested primary isolates and
showed superior potency and breadth to b12, 2F5, 4E10, and 2G12. In addition, scFv-m9 lacks
reactivity to self antigens such as cardiolipin that plagued the development of 2F5 and 4E10. While
effective as a scFv or a Fab, the potency of m9-IgG is significantly less suggesting that the target CD4i
epitope has a size restriction. To exploit scFv m9 or Fab m9 as clinical products, we must extend their
pharmacokinetic half-life in-vivo. Binding to albumin via albumin binding peptides has been used as
a strategy to dramatically improve the pharmacokinetics of antibody fragments without significant
increase in their size. We have engineered two variants of scFv m9, called m9hc and m9sa, in which two
albumin binding peptides independently were fused to the C-terminus of the scFv m9. The new
constructs have been expressed in E-coli and tested for binding to gp120-CD4 complex and for
neutralization of HIV-1. Albumin binding peptide scFv m9 variants preserved the binding and
neutralizing properties of the original scFv m9. If pharmacokinetic characteristics of m9sa and m9hc
constructs are appropriate for once a day injection, these human antibody fragments will be
developed as clinical candidates for anti-HIV prophylactics in incidences of accidental exposure,
mother-to-child transmission; or salvage treatment therapy.
185 Interaction between a domain in the fusion peptide
proximal region (FPPR) of gp41 and the epitope
domain in the membrane proximal external region
(MPER) increases binding of 2F5 to its epitope
Uwe Fiebig, Magdalena Eschricht, Mirco Schmolke, Reinhard Kurth, and

Joachim Denner
Robert Koch-Institute, Nordufer 20, D-13353 Berlin, Germany
Neutralizing antibodies such as 2F5 and 4E10 have been isolated from HIV-1 infected patients.
They recognize epitopes in the MPER of gp41 and neutralize a broad range of HIV strains. Previously we
reported induction of neutralising antibodies against the porcine endogenous retrovirus (PERV), the
feline leukaemia virus (FeLV) and the Koala retrovirus (KoRV) by immunization with the ectodomain of
their transmembrane envelope (TM) proteins p15E. Antisera obtained in rats, goats and cats recognized
two epitopes: One, designated E1, was located in the FPPR and the other was located in the MPER of
p15E (E2). E2 corresponded to the 2F5/4E10 epitope in gp41. Despite the evolutionary distance
between HIV on one hand and PERV, FeLV and KoRV on the other, limited sequence homology exists
(F/YEGWFN in the case of gammaretroviruses and NWFNIT, the 4E10 epitope, in the case of HIV-1,
identical amino acids underlined). Immunising cats with p15E of FeLV resulted in protection from FeLV
infection in vivo.
Using ELISA, epitope mapping and surface plasmon resonance analysis we also identified an E1
domain in the FPPR of gp41 of HIV and showed that the interaction of peptides containing E1
significantly increased the binding of 2F5 to peptides containing the E2 (2F5/4E10 epitope) domain. In
addition, neutralisation of HIV-1 by 2F5 was inhibited more effectively with both gp41-derived
peptides E1 and E2 than with peptide E2 alone.
Since E1 peptides increased the interaction of 2F5 with peptides containing its E2/ELDKWA
epitope, it is most likely that both domains are required to induce neutralizing antibodies (WO
2005/021574, WO 2007/107597). This strategy may be used to generate a vaccine inducing broadly
neutralising antibodies to prevent or curtail HIV infection.
186 Immunosuppressive human endogenous retrovirus
K (HERV-K) is expressed in human villous and
extravillous cytotrophoblasts
Ulrike Kämmerer,1 Ariane Germeyer,2 Michaela Kapp,1 Sven Stengel,3

Kristina Büscher,3 Reinhard Kurth,3 and Joachim Denner3
1
University Women’s Hospital, Wuerzburg, Germany, 2University of Heidelberg, Dept. of Gynecological
Endorinology and Reproductive Medicine, Heidelberg, Germany, and 3Robert Koch Institute, Berlin,
Germany
Overexpression of different human endogenous retroviruses (HERVs) has been shown in stem cell
tumors, melanomas, human embryonic stem cells as well as in the placenta. The envelope proteins of
HERV-W (also known as syncytin 1) and HERV-FRD (syncytin 2) were shown to be involved in cell fusion
allowing to generate the syncytiotrophoblast in the human placenta. One of them, syncytin 2, was
shown to be immunosuppressive, the immunosuppressive activity was associated with its
immunosuppressive (isu) domain. Here we report the expression of another HERV, HERV-K, which
is characterised by open reading frames for all viral genes. Using real time PCR, immunohistochemistry
and Western blot analysis, expression of HERV-K was studied in normal placental tissue of different
gestational ages. The transmembrane envelope (TM) protein of HERV-K was found exclusively in
villous and extravillous trophoblast cells, sparing syncytiotrophoblast and other cells. The expression
increased up to the second trimester and later decreased. This is the first report showing expression of
the TM protein of HERV-K in normal human placental tissue with an exclusive expression in the
trophoblast, suggesting a potential involvement of HERV-K in placentogenesis and pregnancy. In
parallel the immunosuppressive properties of the TM protein and the corresponding isu peptide were
demonstrated. Therefore expression of the immunosuppressive TM protein of HERV-K may contribute
to the protection of the human embryo.
187 Autoresistance to X4 HIV Infection by soluble
suppressor factors secreted from CD4+ T cells
F. Cocchi, A. DeVico, A. Garzino Demo, and R. C. Gallo

Institute of Human Virology, University of Maryland School of Medicine, 725 West Lombard Street,
Baltimore, MD 21201
Control of persistent viral infection relies particularly on cell-mediated immunity comprised of

CD4+ and CD8+ T cells. There is accumulating evidence on the relevance of soluble factor(s) secreted
by CD8+ and CD4+ T cells in controlling HIV replication in vivo. While the a chemokines RANTES, MIP1-
a and MIP1-b collectively account for the suppression of R5 viruses, yet information is lacking on the
identity of the molecules involved in the suppression of X4 viruses. Proteins that inhibit the replication
of X4 HIV isolates were purified from the conditioned media (CM) of immortalized CD8+ and CD4+ T
cell lines from HIV+ long-term non-progressors subjects (LTNPs) and identified as the a chemokines
macrophage-derived chemokine (MDC), thymus and activation-regulated chemokine (TARC) and I309.
These chemokines are secreted primarily by CD4+ T cells but also by CD8+T cells. CD4+ T cells of
asymptomatic HIV+ individuals secreted significantly higher levels of MDC and TARC compared with
subjects who progressed to AIDS. Recombinant human MDC, TARC and I309 induced a dose
dependent inhibition of X4 viruses. A cocktail of neutralizing antibodies against MDC, TARC and I309
abrogated the inhibition of the replication of X4 viruses mediated by the endogenous chemokines in
a dose dependent manner in PBMC and CD8-depleted PBMC cells acutely infected in vitro. These
studies demonstrate that MDC, TARC and I309 represent a major component of the soluble anti-X4
activity of T cells, suggesting that the mechanism whereby CD8+ and CD4+ T cells contribute to the
control of HIV-1 replication may relate to the secretion of these molecules.
188 Targeted delivery of antiHIV siRNAs to T cells in vivo
Premlata Shankar, MD
Department of Biomedical Sciences, Texas Tech University Health Sciences Center, Paul L. Foster School
of Medicine, 5001, El Paso Drive, El Paso, Texas 79905
RNA interference is a promising therapeutic alternative for HIV infection, provided the barrier of
effective delivery to T cells can be overcome. We developed a method for targeted delivery of siRNA to
T cells that relies on a single chain antibody to the pan T cell antigen, CD7 (scFvCD7). The scFV was
conjugated to the positively charged oligo-9-arginine (9R) peptide (scFvCD7/9R) to enable nucleic acid
binding. NOD/SCID IL2rgc-/- mice that are receptive human cell engraftment and support systemic
HIV-1 infection were used as a preclinical model to test the delivery strategy. The chimeric scFvCD7-9R
reagent efficiently delivered siRNA and silenced CD4 gene expression in T lymphocytes in the Hu/PBL
model, where human cell repopulation is by activation and expansion of adoptively transferred
peripheral blood lymphocytes (PBLs). In animals reconstituted with PBLs from normal donors, weekly
treatment with scFvCD7-9R/siRNA complexes targeting a combination of viral genes and the host
coreceptor molecule CCR5, contained viral replication and maintained CD4 T cell numbers after
exogenous challenge with HIV-1. Similar results were also observed with endogenous virus in animals
reconstituted with HIV+ PBLs to mimic established infection. In contrast untreated control mice
displayed a marked reduction in CD4 T cell numbers with significantly higher serum p24 antigen levels.
scFvCD7-9R was also able to deliver siRNA to na¨ve/resting T cells in the Hu/HSC model where the
adaptive immune system was reconstituted from engrafted cord blood CD34+ hematopoietic stem
cells. In Hu/HSC mice, infected with HIVBaL siRNA treatment could control viral replication and CD4 T
cell loss for up to 7 weeks. Thus, we have demonstrated the feasibility of T cell targeted delivery of
siRNA for potential therapeutic applications in HIV infection, using SCID gamma chain ko mice
repopulated with human cells as a novel preclinical model.
189 Defective HIV-1 Proviral Genomes in Natural Viral
Suppressors
L. M. Eyzaguirre, M. M. Sajadi, R. R. Redfield, W. A. Blattner, and J. K. Carr

Institute of Human Virology, University of Maryland School of Medicine, 725 West Lombard Street
Room S 524 Baltimore, MD 21201
Background: A very small proportion of people infected with HIV-1, referred to as Natural Viral
Suppressors (NVS), are able to suppress viral replication to undetectable levels without treatment. One
possible mechanism for this phenotype is inactivation of the virus at the onset of the infection such
that most viral genomes are defective.
Methods: Twenty-nine NVS patients who have been identified from inner city Baltimore have
longitudinal data confirming this phenotype over many years. A group of 18 controls was also selected
(HIV-infected individuals with undetectable viral loads on HAART treatment and those with high viral
loads off of treatment). Nearly full-length proviral genomes were amplified from PBMC DNA and
sequenced; all genes were analyzed for open reading frames, and hypermutation was assessed by
HyperMut software.
Results: The NVS patients were 100% African American, 63% male and 63% injecting drug users.
The controls were similar: 84.6% African American and 69.3% male. Ten of the 29 NVS patients had
proviral genomes that were defective (34.5%): 8 due to G-to-A hypermutation and 2 with frame
shift mutations in either pol or env. Among the 18 controls there were 2 defective genomes (11.1%),
both due to hypermutation. Of note, both of the hypermutated strains were from the 4 controls that
had been on treatment. Most of the hypermutated sites were in a GG context, rather than GA,
implicating APOBEC3G as the enzyme responsible. The mean pairwise genetic distance among the
NVS patients over the entire genome was 5.05% (+/2 1.7%) compared to 6.63% (+/2 1.0%) among the
controls.
Conclusions: A study of 29 NVS patients revealed that 34.5% had defective proviral genomes
compared to 11.1% of the controls. The NVS patients also had less genetic diversity than the controls.
Both of these observations suggest that one of the mechanisms responsible for the NVS phenotype
may be inactivation of the virus early after transmission, leading to the prevention of wide-scale
replication and reduced genetic diversity.
190 The Immune Response to HIV: Friend or Foe
M. Karen Newell,1 Elizabeth Connick,2 Evan Newell,3 Haig Keledjian,3

Monica Ord, 3 Robert Berliner, 3 Joshua Cabrera, 1 Richard Tobin, 1
Cassie Pleasant,1 and Lisa Villalobos-Menuey1
1
University of Colorado, Colorado Springs; 2University of Colorado Health Sciences Center; 3Viral
Genetics, Inc., Asuza, California
Our work aims to investigate a newly discovered link between inflammatory events that
characterize early stages of HIV infection and a link to an infected person’s MHC alleles.
Hypothesis: We propose that it is the inflammation characteristic of an innate immune response
that is responsible for the loss of CD4+ T cells in HIV.
Experiments and Results: We base this proposition on five observations: (1) There is evidence
that, in the absence of functional T regulatory cells (Tregs), the impact of polyclonal B cell activation is
chronic inflammation and the activation of abT cells*; ab T cells have been shown to kill CD4 T cells
during HIV infection**; (2) It has been shown that as T reg numbers decline, there is a concomitant rise
in viral load in the HIV-infected patient*** (3) We have discovered that the B cells from an HIV infected
lymph node have high levels of an important self peptide on their surfaces that indicates the lymph
node B cells have been non-specifically activated. (4) Our data suggests peptide loading of non-
specifically-activated B cells with targeted peptides can stimulate the activity of Tregs in an MHC allele-
dependent manner; and (5) Treatment with a mixture of targeted peptides, reduces viral load in
approximately 25 to 30% of patients treated, a result potentially explained by differences in MHC
between patients.
Conclusions: The consequences of unraveling the link between inflammation, cellular activation,
and MHC may result in understanding a mechanism that may lead to development of targeted
peptides that can redirect the immune response to, and interfere with, the inflammation-dependent
pathology of HIV.
Future Directions: Our exciting findings suggest that, with the proper predictions, we can
synthesize targeted peptides to treat people with all different types of MHC alleles providing an
inexpensive and potentially efficacious biological therapy for HIV.
191 Anti-FasL treatment preserves SIV-specific memory
and slows progression to AIDS in rhesus macaques
Bhawna Poonia, Maria S. Salvato, and C. David Pauza

Institute of Human Virology, 725 W Lombard Street, Baltimore, MD
Previously we reported an increased survival advantage in SIV-infected rhesus macaques after

injection with a FasL-blocking monoclonal antibody (Salvato et al., Clin Dev Immunol. 2007: 93462). In
the present study we analyzed the effect of such FasL blocking on immune parameters and disease
progression in a cohort of SIV-infected rhesus macaques. Upon infection, various immune parameters
in the blood developed highly significant differences between animals given anti-FasL RNOK203
antibody and those given an isotype control antibody. The anti-FasL treated animals had a clear
preservation of central memory CD4 lymphocytes starting from acute infection through at least set
point viremia. The total memory CD4 lymphocytes in these animals were also significantly higher
throughout the 25-week study period. Anti-FasL treatment resulted in significantly higher percentages
of memory CD8 lymphocytes in peripheral blood. Besides these changes in the memory lymphocyte
compartment, the treated animals had a higher response to infection as evidenced by increased
turnover of CD4 and CD8 lymphocytes expressing Ki67. Importantly, the antigen-specific cellular
immune responses were significantly higher at 4, 6 and 17 weeks after infection in anti-FasL treated
animals. Preservation of memory lymphocytes and cellular immunity are considered the main
correlates of protection in SIV infection. Our study demonstrates that FasL is a major contributor
towards depletion of anti-SIV immunity and that blocking this pathway is a beneficial strategy that
should be considered for vaccine formulations.
192 Significant Relationship between INNO-LIAä HIV I/II
Positivity Bands and the Immunological Status of
HIV Patients
Fernanda Leite, Fátima Oliveira, and Luciana Pinho

Clinical Haematology Dept. -Hospital Geral Santo Antonio- 4050 Porto-Portugal
Objective: the aim of our work was to study the relation between the INNO-LIAä HIV I/II (ILH)
positivity band patterns and the immunological status of HIV patients.
Methods: We looked for HIV patients whose ILH was performed between June 2007 and June
2008. The ILH bands were classified after the grade of positivity of the different antibodies: anti-
sgp120, anti-gp41, anti-p31, anti-p24, anti-p17, anti-sgp105 and anti-gp36. We looked for age, gender,
n° total leucocytes, n° total lymphocytes, n° of CD3+, CD4+, CD8+, CD19+, NK cells and HIV viral load.
We looked also for HCV, HBV and HAV status. We studied 247 individuals, 65 were female (26,3%) and
182 male (73,7%) with ages between 15 and 76 years old with mean age of 40. Five patients were HIV-2
patients. Data statistical analysis was performed with SPSS program and P value ,0.05 was regarded
significant.
Results: Sgp120 positivity was significantly related with the decreasing lymphocyte number (P =
.046) and in the linear regression equation the HAV independent variable showed the highest
predictive value for sgp120 positivity (P = .031). Gp41 was significantly related with HIV viral load (P =
.040) and with HCV co-infection (P = .005); HCV and HAV were independent variables with predictive
value for gp41 positivity (P = .001). P31 was associated significantly with the decreasing n° of
leucocytes (P = .043) and CD19+ cells (P = .011); HIV viral load had the highest predictive value for p31
positivity.
The positivity of p24 band was associated with increasing number of CD4+ (P = .0.000) and CD19+
cells (P = .000) and decreasing CD8+ cells (P = .001). There wasn’t any significant correlation between
p17, sgp105 nor gp36 and immunological parameters. Women showed higher CD4+ cells (P = .048)
and p24 positivity (P = .010), being the gender the independent variable with the highest predictive
value for the p24 positivity (P = .007). Men had higher HIV viral load (P = .019) and higher incidence of
HCV co-infection (P = .021). CD4 cells decreased with age (P = .015). NK cells showed a decrease till 60
and increased afterwards (P = .000). HCV incidence was higher till 49 (P = .000), being 38, 8% of the
population HCV co-infected. HBV carrier status was presented in 5, 8% and 40, 2% of the population
had HBV resolved infection. HAV IgG was presented in 85, 8% of the individuals. CD4+ cells was
significantly associated with HIV viral load (P = .000) and CD19+ cells (P = .000).
192 Significant Relationship between INNO-IAä HIV I/II
Positivity Bands and the Immunological Status of
HIV Patients
(Continued)
Discussion and Conclusions: In spite of the limitations of a cross-sectional study, we found
a significant relation between INNO-LIAä HIV I/II positivity bands and immunological parameters in
HIV infected patients. Anti-sgp120, anti-gp41 and anti-p31positivity are associated with a more
immunodepressed status and a higher HIV viral replication. Concerning p24 antibody: is there
a protective role for women? Is there a role for anti-p24 in vaccine issue? HCV co-infection and past
HAV infection are associated with poorer immunological parameters. This issue highlights the
importance of the chronic immune activation as the major factor for AIDS progression. The correlation
found between CD4+ cells, CD19+cells and HIV viral load highlights the importance of the study of B
cells in the pathogenesis and therapy for HIV infection.
193 The HIV-Positive Inpatient: Psychosocial Risks and
Adherence Implications
Rebecca L. Wald, Stephen J. Synowski, and Lydia R. Temoshok

Institute of Human Virology and Department of Medicine, University of Maryland School of Medicine,
Baltimore, MD, USA
Prior research on adherence to anti-retroviral therapy (ART) has focused on correlates of

adherence among outpatients. Little is known about adherence among hospital inpatients with HIV, or
about the prevalence of psychosocial factors associated with nonadherence in inpatient populations,
despite the fact that their poorer health status typically denotes a critical need for ART initiation and
adherence.
200 HIV-positive inpatients and 200 outpatients (52% male, 95% African-American) completed
a structured interview assessing nonadherence to ART and psychosocial variables associated in
previous research with nonadherence. Inpatients reported missing significantly more doses of ART in
the week prior to hospitalization than did outpatients (t = 5.67, P , 001). Inpatients had poorer scores
on measures of multiple psychosocial predictors of nonadherence: social instability (t = 23.42, P =
001), depressive symptoms (t = 23.61, P , 001), other psychiatric symptoms (t = 2.95, P = 003), recent
heroin or cocaine use (x2 = 25.2, P , 001) and cognitive dysfunction (t = 22.76, P = 006), and reported
life stressors of greater severity (t = 3.99, P , 001). Inpatients were significantly less likely to have
a regular HIV doctor (x2 = 37.2, P , 001) or to have seen an HIV specialist in the 6 months prior to
hospitalization (x2 = 74.37, P , 001); those in care reported lower satisfaction with their provider (t =
217.34, P , 001).
In summary, chronic and severe psychosocial problems found in prior research to be associated
with nonadherence are widespread among HIV-positive inpatients. Inpatients also display a substantial
lack of engagement in HIV care. Adherence studies conducted with outpatients do not adequately
reflect the difficulties of this population. Most inpatients will require intensive psychosocial
interventions in order to achieve ART success.
193a Psychosocial Contributors to Antiretroviral
Adherence: Stability and Change
Rebecca L. Wald, Stephen J. Synowski, and Lydia R. Temoshok

Institute of Human Virology and Department of Medicine, University of Maryland School of Medicine,
Baltimore, MD, USA
Adherence to anti-retroviral therapy (ART) is critical to HIV treatment success, yet many fail to
achieve the .95% adherence necessary for viral suppression. This study examined the stability of
psychosocial predictors of adherence.
127 HIV+ adults (92% African-American, 51% female, mean age 44.5), enrolled in a longitudinal
study of HIV progression and receiving ART, completed a structured interview assessing nonadherence
and related psychosocial variables at baseline and at 6- and 12-month follow up. The Hardiness Scale
(HS), Perceived Stress Scale (PSS), and a coping measure (Vignette Similarity Rating Method; VSRM)
were also completed. Patients were classified as adherent if they took .95% of ART doses in the past
week.
Adherence was stable from baseline to 6-month (O2 = 15.86, P , 01) and 12-month (O2 = 8.31, P
, 01) follow-up. At each point, different psychosocial problems were associated with concurrent
nonadherence: at baseline, depression (t = 2.13, P = 04), psychiatric symptoms (t = 2.23, P = 03),
and interviewer-rated stress severity (t = 2.63, P = 01); at 6 months, stress severity (t = 2.36, P = 02) and
hopelessness (VSRM) (t = 2 1.88, P = 07); and at 12 months social support (t = 22.33, P = 02),
unsupportive social interactions (t = 2.073, P = 042), alcohol use (t = 22.86, P , 01), stress severity (t =
1.875, P = 06), and PSS scores (t = 1.863, P = 06). Being adherent at follow-up was associated with
baseline coping variables: higher scores for optimism (t = 2.86, P , 01); adaptive coping (VSRM)
(t = 22.07, P = 042); and the Commitment (t = 22.51, P = 02) and Control (t = 22.15, P = 04) subscales
of the HS, which respectively assess a sense of purpose and a sense of autonomy.
Nonadherence in this sample was stable over time, but the associated acute psychosocial
problems differed at each time point. The stability of adherence appears to be driven by more stable,
trait-like coping patterns.
194 Systolic Blood Pressure recovery following mental
stress predicts immune dysregulation in persons
with HIV
Stephen J. Synowski, PhD

725 West Lombard Street, Baltimore, Maryland 21201
Cardiovascular reactivity (CVR) to, and prolonged recovery from, mental stress have been
associated with impaired immune function. In 128 HIV-infected patients enrolled in a longitudinal
study of potential mechanisms of HIV progression, baseline analyses revealed that exaggerated CVR
and a poorer recovery were associated with decreased production of the HIV-inhibiting beta-
chemokines MIP-1a/b (which block the HIV co-receptor CCR5 and thus, HIV entry into CD4+ T-cells).
We hypothesized that these relations would be part of a chronic pattern of psychophysiologic
dysregulation. At 6-month follow-up, participants were 95 HIV+ adults (92% African-American, 51%
female, mean age 44.5). At baseline, systolic and diastolic blood pressure (SBP, DBP) and heart rate (HR)
were monitored during emotion-provoking anger recall and role play tasks; each task was preceded by
10 minutes of rest and followed by 5 minutes of recovery. At 6 month follow-up, in vitro production of
MIP-1a/b was assessed as stimulated by the core HIV protein p24. Supernatants were collected on
days 3 and 6, and assays performed by ELISA. Poorer recovery to resting SBP levels following role play
(at baseline) predicted decreased 6-month production of MIP-1a (b = 20.21, P , 044, R = 0.74) and
MIP-1b (b = 20.26, P , 016, R = 0.63). Additionally, the relations between SBP reactivity (b = 20.18,
P , 093, R = 0.67) and poorer recovery (b = 20.19, P , 079, R = 0.64) following anger recall and
decreased MIP-1b production approached statistical significance. In sum, prolonged SBP recovery
from mental stress at baseline significantly predicted decreased HIV-specific immune response at 6
month follow-up. These findings are consistent with previous results obtained at baseline, and suggest
that a chronic pattern of psychophysiological dysregulation may be associated with suppression of
anti-HIV b-chemokine production, and thus, may be implicated in HIV progression.
195 Stimulation of reverse cholesterol transport
potently suppresses HIV-1 replication
Michael Bukrinsky
The George Washington University, 2300 Eye St. NW, Washington, DC 20037
Cholesterol plays an important role in HIV life cycle, and infectivity of cholesterol-depleted HIV
virions is significantly impaired. Recently, we demonstrated that HIV-1, via its protein Nef, inhibits
activity of the cholesterol transporter ABCA1 and impairs cholesterol efflux from infected
macrophages. However, the role of this pathway in HIV infection of CD4+ T cells remained unclear.
In this study, we investigated the effect on HIV replication of an LXR agonist, TO-901317, which is
a potent inducer of ABCA1 expression and of reverse cholesterol transport. Treatment with TO-901317
restored cholesterol efflux from HIV-infected macrophages and CD4+ T lymphocytes, and potently
suppressed HIV-1 replication in both cell types. The drug also inhibited HIV-1 replication in ex vivo
cultured lymphoid tissue and in humanized Rag2/2gc2/2 mice. Cholesterol analysis demonstrated
a significant enrichment with cholesterol of membrane rafts in cells infected with wild-type, but not
Nef-deficient, HIV-1. Stimulation with LXR agonist reduced raft cholesterol in both infected and
uninfected cells. The amount of virion-associated cholesterol correlated with raft cholesterol: Nef-
deficient virus had less cholesterol than the wild-type virus, and cholesterol in both types of virions
was reduced by LXR agonist. Importantly, infectivity of virions closely matched the amount of virion-
associated cholesterol. These results demonstrate an extreme dependence of HIV infectivity on activity
of reverse cholesterol transport and describe a novel approach to reduce HIV virion cholesterol. They
suggest that drugs targeting LXR-ABCA1 pathway, in addition to their anti-atherogenic activity, are
potential anti-HIV agents working through a novel mechanism of action. Such drugs may provide
a double benefit to HIV-infected patients by reducing HIV replication and preventing development of
atherosclerosis associated with HIV infection.
196 Monocyte-Dependent and –Independent
Modulatory Effects of Vitamin D3 on X4 and R5 HIV-1
Replication in IL-2 Stimulated PBMC
Massimo Alfano,1 Chiara Rizzi,1 Giuseppe Penna,2 Luciano Adorini,2 and

Guido Poli1,3
1
San Raffaele Scientific Institute; 2BioXell SpA; 3Vita-Salute San Raffaele University, School of Medicine,
Via Olgettina n. 58, 20132 Milan, Italy
Background: Vitamin D3 (calcitriol) is a well known mononuclear phagocyte differentiating agent

as well as a potent modulator of the immune system. We investigated the potential role of Vit D3 on
the replication of R5 and X4 HIV-1 strains in primary PBMC stimulated with IL-2 in the absence of PHA,
a model system in which virus replication is dependent upon monocyte-T cell interaction and release
of endogenous cytokines.
Methods: IL-2 PBMC were incubated with Vit D3 or vitamin D receptor (VDR) agonists like calcitriol
analog BXL219 and infected with laboratory adapted or primary X4 and R5 HIV-1 strains. Virus
production and cytokines levels were measured in culture supernatants, chemotaxis and the levels
of CD4, CCR5 and CXCR4 were determined by cytofluorimetric analysis.
Results: VDR agonists enhanced laboratory-adapted and primary R5 and X4 HIV-1 replication in IL-
2 PBMC. Unlike R5 HIV-1, X4 virus spreading was initially inhibited by VDR agonists in the first 2-3
weeks of infection, as determined by virion-associated RT activity in cell culture supernatants. Neither
the levels of CD4, CCR5 or CXCR4 expression nor the chemotactic response to their ligands CCL4 and
CXCL12 were affected by calcitriol. Neither R5 nor X4 HIV-1 infection affected significantly the
production of pro-inflammatory cytokines (TNF-a, IL-1b, IFN-g and IL-6), IL-1ra and chemokines (CCL3,
CCL4, CCL5, CXCL8 and CCL11) by IL-2 PBMC. Calcitriol significantly inhibited the production of TNF-a,
IFN-g and CCL5, but not of the other tested molecules, in both infected and uninfected cells. Of
interest, X4 infection did not modify calcitriol effect on TNF-a and IFN-g whereas it abrogated the
inhibitory effect induced by calcitriol on CCL5 expression. Stimulation of IL-2 PBMC with calcitriol
enhanced the differentiation of monocytes into adherent macrophages. The replication of both R5
and, remarkably, X4 HIV-1 was greatly enhanced in adherent IL-2 PBMC (mostly monocyte/macroph-
ages) upon stimulation with VDR agonists whereas non-adherent lymphocytes showed patterns of
virus replication superimposable to those of unfractionated IL-2 PBMC.
Conclusions: VDR agonists predominantly enhance HIV replication in both lymphocytes and,
particularly, monocyte/macrophages regardless of which chemokine entry co-receptor is used in
PBMC stimulated with IL-2 suggesting caution in their clinical use in HIV-infected individuals.
197 Naturally Occurring C-terminally Truncated Isoform
of STAT5 (STAT5D) Is a Transcriptional Repressor of
HIV-1 Expression. Characterization of the Platinum-
Based STAT Inhibitor CPA-7
Giulia Della Chiara,1 Andrea Crotti,1 Heidi Kay,2 and Guido Poli1
1
AIDS Immunopathogenesis Unit - San Raffaele Scientific Institute, Milano, Italy; 2College of Public
Health, University of South Florida, Tampa, Florida
Signal transducers and activator of transcription (STAT) proteins, namely STAT1 and STAT5D, are
frequently constitutively activated in the PBMC of most of HIV-1+ individuals. We previously described
that STAT5D is also the only STAT5 isoform detectable in the chronically HIV-1 infected promonocytic
cell line U1 characterized by a constitutive state of viral latency and inducibility of virus expression by
PMA or several cytokines. We have recently reported that STAT5D can act as inhibitor of HIV-1
transcription in GM-CSF-stimulated U1 cells and IL-2 stimulated PBMC of HIV+ individuals due to its
binding to target DNA sequence in the provirus LTR causing an impaired recruitment of RNA Pol II
(A. Crotti et al., Blood, 2007). GM-CSF also triggered the late activation of an ERK/AP-1 dependent
pathway inducing HIV-1 expression in U1 cells. Selective inhibition of this pathway turned off, while
inhibitors of STAT5 enhanced, viral expression in GM-CSF stimulated U1 cells. We are currently investi-
gating a new platinum-based compound, termed CPA-7, reported to interfere with STAT3 in several
solid tumor cell lines by inducing apoptosis. Our preliminary results demonstrate that CPA-7 increased
HIV-1 expression in GM-CSF stimulated U1 cells. Further investigation will clarify the role of this
compound in the context of STAT-induced HIV-1 expression and will suggest a possible therapeutic use
of CPA-7 in the context of HIV reactivation from latently infected cells.
198 Modulation of CCR5 density with low doses of the
transplant drug Rapamycin sensitizes Vicriviroc-
resistant R5 HIV-1
Alonso Heredia
Institute of Human Virology, Rm N549, 725 W. Lombard Street, Baltimore, MD
CCR5 antagonist (CA) are a new class of anti-HIV drugs. With a novel mechanism of action, they
offer additional treatment options to patients, especially those infected with drug-resistant HIV-1.
Maraviroc is the first approved CA. Vicriviroc (VCV), another CA, is undergoing evaluation in Phase III
clinical trials. We show that reduction of CCR5 density (CCR5 receptors/cell) with low doses of the
transplant drug Rapamycin (RAPA) synergistically enhances VCV antiviral activity (see Abstracts by
Latinovic et al.). We have also evaluated the antiviral activities of RAPA and RAPA/VCV combination
against D1/85.16 and CC101.19, two VCV-resistant R5 HIV-1 strains that use CCR5 but can also use VCV-
bound CCR5. RAPA inhibited these viruses by up to 60%, in agreement with their dependence on CCR5
for infection. Importantly, RAPA partially sensitized these viruses to VCV, with the RAPA/VCV
combination inhibiting viral production in lymphocytes by ;90%. We further demonstrate that
resistance to VCV in IL-2 primed lymphocytes depends on donor CCR5 density, with lower densities
partially inhibiting virus replication. We confirmed the effect of CCR5 density on VCV resistance in
pseudovirus infection of cell lines with same CD4 but varying CCR5 densities. Titration of VCV against
VCV-resistant pseudovirus yielded incomplete inhibition curves, with inhibition plateau heights
inversely correlated to CCR5 density. In T-20 time-of-addition experiments, VCV-resistant pseudovirus
infection kinetics were considerably slower in VCV-bound than in CCR5 free usage. These results
indicate that VCV-resistant HIV uses VCV-bound CCR5 less efficiently than free CCR5 and suggest that
the affinity of the VCV-resistant envelope is lower for bound CCR5 than for free CCR5. An alternative
explanation to differences in affinity is that the energy requirements for induction of conformational
changes in gp41 following CCR5 binding are higher in use of VCV-bound CCR5 than in use of free
CCR5. Together, these results suggest CCR5 modulation with low doses of RAPA as a potential
approach to prevent and control resistance to VCV in patients.
199 Risk factors for Virologic Failure and Adverse
Reactions among Patients on Triple Antiretroviral
therapy
Adedayo Adeyemi, Oluseyi Adesola, and Oluyemisi Olaogun

Healthmatch International, Lagos, Nigeria
Introduction: Adverse effects from antiretroviral (ARV) drugs can result in poor compliance and
which may results in treatment failure. Recognition and management of adverse effects are important
issues in improving quality of lives of HIV positive patients. This study examined the prevalence of
adverse effects and the risk factors associated with adverse effects and virologic failure.
Methods: Retrospective chart review of 146 naive patients on antiretroviral therapy between January
2007 and December 2007 was done in Lagos Nigeria. The following information was obtained: time of first
diagnosis, demographical information, missing clinic, CD4 count, weight, tuberculosis/opportunistic
infections, viral loads (at least one in 6 months after starting ARV), and partner/family notification.
Results: Female 93 (64%), male 53 (36%); mean age: 33.4 years; 234 person-years of follow-up with
a mean of follow-up of 1.6 6 0.7 years; 126 (86%) had undetectable viral load at 6 months of ARV;
mean weight change from 56 kg to 62 kg; and 53 (36%) experienced adverse effects. Adverse effects
were peripheral neuropathy 35 (24%); gynecomasia 6 (4%) lactic acidosis 10 (7%); diarrhea 18 (12%);
nausea and vomiting 4 (3%); hepatitis 6 (4%); Stevens-Johnson syndrome 3 (2%); rash 20 (14%);
insomnia 12 (8%); lipodystrophy/lipoatrophy 9 (6%). CD4 count and Viral load at initiation were 2
significant predictors of virologic failure, with CD4 , 100 cells/mm3 (OR 1.6, 95% CI 1.2 to 1.9) and viral
load at initiation $100,000 copies/mL (OR 2.1, 95% CI 1.5–2.7). For the adverse effects: patients with
weight ,60 kg (OR: 3.2, 95% CI 1.9 to 4.2) and with age .40 years OR 2.4 (1.2 to 3.9) are more likely to
experience adverse effects. Mean change of regimen was 13.5 6 6.2 months.
Conclusion: Despite increasing access to ARV, good drug toxicity monitoring system should be
put in place so that life threatening adverse effect is adequately screened for and proper managed.
However, care should be taken with D4T associated peripheral neuropathy. More surveillance studies
on HIV drug toxicity are needed to enhance safety and effectiveness of the therapy in Nigeria.
200 Design, Synthesis, Anti-HIV and Cytotoxicity of
Novel Heterocyclic Compounds
Periyasamy Selvam
Amrita School of Pharmacy, Elmakkara, P.O. Kochi, 682026, Kerala, India
AIDS is a fatal pathogenic diseases caused by retrovirus HIV. Recently much attention has been
devoted for searching of effective chemotherapeutic agents for combat HIV/AIDS. Present work is to
design and synthesis of novel heterocyclic compounds form Indole, Benzoxazine, Quinoxaline,
Quinazolinone, Flouroquinolone, Phthalimide, benztriazole and benzimidazole lead molecules and
characterized by spectral analysis. Synthesized compounds were screened for in vitro antiviral activity
against HIV-1 & 2 in MT-4 cells. Cytotoxicity is also investigated in uninfected MT-4 cells by MTT assay.
From the results of anti-HIV activity, 2,3-diphenylquinoxaline and 3-sulphonamido-quinazolinones
inhibits and sulphadimidinylmethyl benzimidazole replication of HIV-1 and 2. Benztriazole and
Benzimidazole derivatives displayed marked cytostatic activity in MT-4 cells. Details of Design,
Synthesis, anti-HIV activity and cytotoxicity will be presented.
200a Inhibition of HIV replication and integrase activity
by isatin derivatives
Periyasamy Selvam
Amrita School of Pharmacy, Elmakkara, P.O. Kochi, 682026, Kerala, India
We evaluated the broadspectrum antiretroviral activity of some novel isatin (SPIII) and its
derivatives using different strains of HIV-1, HIV-2 and SIV. Several SPIII derivatives were found to inhibit
HIV-1 (IIIB and NL4.3) replication in MT-4 cells at EC50 values ranging from 3.6 to 49.6 mg/mL. No
cytotoxicity was observed at concentrations up to 125 mg/mL in mock-infected MT-4 cells. The
compounds were evaluated for their inhibitory effects against a variety of mutant HIV strains. The SPIII
derivatives showed low level resistance (up to 11-fold) to NL4.3 virus strains resistant to agents that
block virus entry, i.e. NL4.3 strains resistant to the binding inhibitor dextran sulphate or the CXCR4
antagonist AMD3100. Moreover, the SPIII-5Br and 5Cl derivatives showed some low level cross-
resistance to an integrase inhibitor resistant virus. We observed no cross-resistance against the fusion
inhibitor, nucleoside reverse transcriptase, nonnucleoside reverse transcriptase or protease inhibitor
resistant HIV strains. The SPIII compounds were found to inhibit the human immunodeficiency virus
type 1 integrase enzymatic activity, HIV-1 integrase binding to target DNA and adsorption of HIV-1 to
MT-4 cells, all at IC50 values in the mg/mL range. We conclude that it would be interesting to
synthesize and evaluate more derivatives of SPIII to identify more selective congeners. These
compounds would enable us to determine the structural requirements for inhibition of entry or
integration steps in the replication cycle of HIV.
201 Pathogenesis induced acidosis and hpercalceamia
evident in HIV/AIDS disease
Abdulrazak Hamza Yahaya

HIV/Immunology Laboratory, Pathology Department Murtala Muhammad Specialist Hospital, Kano,
Nigeria
Background: Metabolism is essential to functioning of cellular systems. HIV destroys the host
immune system and induced metabolic changes that interfere with their normal functions. It is
imperative to control the induced changes caused by the viral disease while rehabilitating the host
immune system through therapeutic care.
Methods: Serum electrolytes and calcium were studied among eighty volunteers, 50% HIV
infected and 50% were uninfected. Volunteers’ bio data were obtained using structured
questionnaires and blood samples were collected and tested for HIV antibodies and electrolytes
data were analyzed by SPSS v 12.0.
Results: Study observed marked variation in mean values of electrolytes between HIV infected
and uninfected volunteers. Respective serum concentration of Calcium, bicarbonate and potassium
recorded in infected and uninfected groups were 35.9 6 16.8 mg/dL, 18.6 6 3.8 mMol/L, 4.45 6 1.0
mMol/L and 10.4 6 1.0 mg/dL, 29.7 6 1.0 mMol/L, 3.9 6 0.9 mMol/L. HIV pathogenesis induce changes
in host system, most interesting are those related to immune coordination and metabolism; observed
alteration in calcium and bicarbonate in HIV/AIDS patients studied are significant at P , 0.01.
Conclusions: Great deal of care is required for HIV/AIDS patients to have a quality life. Finding
from our biochemical evaluation results showed variation in serum bicarbonate and calcium level
among infected cohorts as compared with uninfected cohorts, thus there is need for correcting these
abnormalities in HIV/AIDS disease. Normal hosts’ metabolisms required for correct cellular processes
and safety from drug toxicity can be altered by electrolytes imbalance.
Key Words: alteration; electrolytes; HIV
202 Retinopathy and enteropathy in HIV patients,
a hypothetical concept beyond Virus and CD4 levels-
a study from Varanasi, North India
Dr. V. Satya Suresh Attili, Prof, Shyam Sundra, Prof, V. P. Singh, Prof, and
Prof, A. K. Gulati
Room No 20, Yashoda Hospitals, Somajiguda, Rajbhavan Road, Hyderabad-500082
Background: Based on our previous observations which were published (Asian journal of
ophthalmology, 2006. July p341-48 and BMC Infectious Diseases 2006, 6:39) we thought of re
evaluating the results of the hospital based cohort to propose possible hypothesis and evaluate it in
prospective manner.
Aim: To evaluate the effect of local immunity on development of HIV retinopathy and
enteropathy in a hospital based cohort at Institute of Medical Sciences, Banaras Hindu university,
Varanasi, India.
Methods: All the patients with HIV retinopathy or enteropathy (diagnosis of which was
established by ruling out all other pathologies like opportunistic infections, malignancies in cases of
enteropathy and infections, malignancies and other confounders like hypertension, diabetes in
retinopathy cases) were enrolled and equal number of controls, who had similar symptoms during the
same time was recruited for comparison. The baseline characters, CD4 levels, duration of symptoms,
number of episodes and other possible confounders were compared in the groups.
Results: The multivariate analysis for the retinopathy suggested that the local inflammation &
immunity are most important risk factors as significant number of patients have near normal
circulating CD4 counts. All retinopathy patients had some form of local infection (which indirectly
indicated a decreased local immunity) which further supports our hypothesis. Similarly in patients with
enteropathy, we found that around 78% of chronic and 45% of acute diarrheal episodes were
experienced by patients with CD4 counts of .200 cells/mL, indicating a regional immunosuppression.
Similar observations were made earlier (GUT. 1995;37:524–29) where authors observed that, loss of
CD4 cells in intestinal mucosa of the patients with diarrhea, were more pronounced than peripheral
CD4 levels and their relation is quite variable.
Conclusions: The local immunity, an important factor to prevent retinopathy as well as enteropathy
may therefore be variable and therefore they can be observed even in patients with good immunity (i.e.
peripheral CD4 . 200). However, it would be premature to draw definitive conclusions beyond this, as
the present study is retrospective one and number of patients analyzed was relatively small.
203 Immunosuppression Level In HIV-1 –Infected
Patients Doesn’t Affect the Serological Diagnosis of
Hepatitis C Virus
Fernanda Leite, and Luciana Pinho

Clinical Haematology Dept- Hospital Geral Santo Antonio- Porto- Portugal
Objective: The aim of our work was to study immunological variables in RIBAÒHCV 3.0 results in
HIV-1/HCV Co-Infected Patients (CIP).
Methods: we retrospectively looked for HIV-1/HCV CIP with RIBAÒHCV 3.0 Indeterminate (Group 1)
and RIBAÒHCV 3.0 Positive (Group 2), between 1/2002 and 1/2007. We looked for age, gender, n° total
leucocytes, n° total lymphocytes, n° of CD3+, CD4+, CD8+, CD19+, NK cells and HIV-1 viral load. We
studied 87 individuals, 41-Group 1 (47, 1%) and 46- Group 2 (52, 9%); 17 were female (19, 5%) and 70
male (80, 5%) with ages between 1 and 69 years old with mean age in the subgroup 30–39 years. Two
individuals were excluded from the study because data wasn’t coincident in time. Data statistical analysis
was performed with SPSS program and P value , 0.05 was regarded significant.
Results: The 2 groups didn’t show differences statistically significant considering age. In Group 1
there wasn’t a significant relation between the HIV-1 viral load and CD4 cells in contrast with Group 2
that showed an inverse relation between these two variables (P = .040). In Group 2 the n° of CD4+ cells
was lower than Group 1 (P = 0.49) with mean CD4+ cells in total population of 407/il. We found
a significant positive relation between CD19+ cells and CD4+ cells in all population (Group 1- p. = .006;
Group 2-p. =.003). We didn’t find significant differences in the parameters studied in the two Groups;
except when CD4 were inferior to 50 cells/il (n = 7), Group 1 showed lower CD3+ cells than
Group 2 (P = .029).
Conclusions and Discussion: We didn’t find significant differences between the two Groups
considering the immunological variables and HIV-1 viral load. The number of CD4+ cells didn’t
influence RIBA HCV indeterminate results, in spite of only 7 individuals had less than 50 T CD4+ cells.
We hypothesize there’s a higher decrease in circulating memory B cell population in the HIV-1/HCV co-
infected RIBA indeterminate Group since very early in the infection occurs a preferential depletion of
splenic marginal zone-like peripheral blood CD27+B220- memory cells in HIV-1 individuals
after M. Morrow et al. (2006) not related with CD4 cells nor viral load what may contribute to the
impaired immunity response against pathogenic antigens. The positive significant relation found
between CD19+ and CD4+ cells highlight the importance of the study of B cells in the pathogenesis
and treatment of HIV disease.
204 Serum albumin: Could it be an inexpensive and
simple marker of immunosupression in HIV-infected
individuals?
Muthu Sundaram
YRG Centre for AIDS Research and Education (YRG CARE), Voluntary Health Services Campus, Chennai
600 113, India
Background and Aims: Several studies have suggested that albumin could serve as a useful
marker of HIV disease progression in resource-limited settings. We identified the relationship between
serum albumin and absolute CD4+ cell count in subjects attending a HIV referral centre in Chennai,
Southern India.
Methods: In a cross-sectional study, 200 HIV-1 infected individuals were studied. The absolute
CD4+ cell counts were determined using Guava Personal Cell Analyser (Guava Technologies Inc.,
Hayward, CA, USA). Serum albumin was estimated by Bromo Cresol Green method (BCG) using
Olympus AU 400 clinical chemistry autoanalyzer (Olympus Optical, Japan).
Results: We observed that the correlation co-efficient of the serum albumin assay as compared
with the absolute CD4+ cell count was 0.90 (CI:058–0.92; P = 0.0002). Subjects that had absolute CD4+
cell count ,200 cells/mL had a better correlation (R = 0.83, 95% CI:0.72–0.92; P = 0.001) than those with
200–350 cells/mL (R = 0.82, 95% CI:0.72–0.93; P = 0.001) and .350 cells/mL (R = 0.71, 95% CI:0.52–0.80;
P = 0.106). The sensitivity and specificity between serum albumin and absolute CD4+ cell ,200
cells/mL were 97% and 44% respectively.
Conclusions: Serum albumin was strongly associated with CD4+ cell count. However, this
warrants further investigation on their possible application as a useful marker of HIV disease
progression and monitoring of treatment outcome in resource-limited settings.
205 Increased IFN-alpha expression in circulating
plasmacytoid dendritic cells of HIV-1 infected
patients despite selective loss
Clara Lehmann, MD,1,3 Dirk Taubert, PhD,2 Jill M. Harper, PhD,3

Norma Jung, MD,1 Pia Hartmann, MD,1 Gerd Fätkenheuer, MD,1 and
Fabio Romerio, PhD3
1
First Department of Internal Medicine, University of Cologne, Cologne, Germany; 2Department of
Pharmacology, University of Cologne, Cologne, Germany; and 3Institute of Human Virology, University
of Maryland School of Medicine, Baltimore, MD
Background: The role of plasmacytoid dendritic cells (PDC) and interferon alpha (IFNg) in HIV-1
infection is still unclear. On one hand, HIV-1 disease is associated with a progressive decline of pDC,
which display reduced ability to produce IFNg after in vitro challenge. On the other hand, high IFNg
serum levels in HIV-1 infected individuals have been proposed to promote immune hyper-activation
and disease progression. Further, it remains unknown whether distinct IFNg subtypes are expressed
with disease progression.
Methods: We sought to determine whether disappearance of pDC in HIV-1 disease is due to
homing in lymphoid tissues. We also studied IFNg and MxA expression as well as the profile expression
levels of twelve known IFNg subtypes in unstimulated pDC, and correlated these results with selected
clinical and laboratory parameters in PBMC from normal donors and HIV-1 patients at CDC stage A and
stage C of the disease.
Results: PDC decline markedly in peripheral blood of HIV-1 patients progressing to disease, but at
the same time express much higher levels of IFNg and MxA compared to control individuals. On the
other hand, steady pDC counts in lymph nodes of HIV-1 patients are observed, which correlated
directly with CD4 cell counts and inversely with viral load. However, no correlation between IFNg and
MxA expression levels, CD4 counts, and viral load were found. The analysis of IFNga subtype
expression pattern demonstrated high IFNg2 and 6 mRNA levels in both patient groups. Especially,
IFNg2 was upregulated .60-fold in stage A and .400-fold in stage C patients compared to controls,
which correlated with declining CD4 counts. The subtypes IFNg1/13, 8, 14, 16, 17 and 21 were
upregulated in stage C but not stage A patients. The other IFNg subtypes did not change among the
three groups.
Conclusions: Circulating pDC decline sharply in the course of HIV-1 disease, but express high
levels of IFNg. Furthermore, distinct IFNg subtypes are sequentially activated during HIV-1 infection,
which may be predictive of disease progression.
206 HIV-Induced Alterations in Vg2Vd2 T-cell Phenotype
and Function Impact Mechanisms for Tumor
Immunity in AIDS
Jean-Saville Cummings
IHV- Room s507A, 725 W. Lombard St, Baltimore, MD 21201
Human Vg2Vd2 T-cells represent 3-10% of CD3+ lymphocytes and are known to function in innate
tumor surveillance. Several studies have demonstrated that HIV infection causes rapid and lasting
defects in the population of Vg2+ cells, specifically describing a reduction in the overall frequency of
circulating Vg2Jg1.2-Vd2 T-cells. In an effort to further describe the impact of HIV on these cells, we
examined PBMC samples from HIV+ patients receiving HAART who had displayed prolonged viral
control and CD4 counts above 300 cells/mm3. Among HIV+ individuals we observed lower frequencies
of CD27-/CD45RA- Vg2Vd2 cells when compared to uninfected controls. Vg2+ T-cells from HIV+
donors had lower expression of granzyme B and displayed reduced cytotoxicity against Daudi cell
targets after in vitro stimulation. Additionally, the frequency of tumor reactive CD56+ Vg2 effectors
was reduced among HIV+ individuals when compared to controls. Similar reductions in CD56+
frequency were observed among CD3- NK cells and CD3+Vg2- aa T cells within the HIV patient group.
There was increased expression of Fas receptor (CD95) on Vg2 cells from HIV+ PBMC, which may be
a mechanism for depletion of Vg2 cells during disease. In addition to the well-characterized defects in
Vg2 repertoire and functional responses to phosphoantigen, the proportion of CD27-/CD45RA-
Vg2Vd2 T-cells after IPP stimulation was reduced sharply in HIV+ donors versus controls. Similarly, the
CD56+ frequency of Vg2+ cells generated after PBMC stimulation in vitro was also reduced among
HIV+ individuals. None of the differences observed among HIV+ donors were related to CD4 count or
vRNA burden. Thus, HIV infection has multiple impacts on the circulating Vg2Vd2 T-cell population
that combine to reduce the potential effector activity in terms of tumor cytotoxicity. Changes in
Vg2Vd2 T-cells, along with concomitant effects on NK and NKT cells that also contribute to tumor
surveillance, may be important factors for elevating the risk of malignancy during AIDS.
207 Mechanisms regulating Vg2Vd2 tumor cell
cytotoxicity: Roles for CD56 and T cell receptor
Kun Luo
Institute of Human Virology University of Maryland, 725W Lombard Street, Baltimore, MD 21201
Human Vg2Vd2 T lymphocytes respond to stimulation with non-peptidic phosphoantigens plus

IL-2 and become cytotoxic for a variety of human tumor cell lines. Recently, we reported that CD56
expression marked a subset of activated Vg2Vd2 T cells that were most potent in cytotoxicity assays
(Alexander, et al., Clin. Cancer Res. 14: 4232, 2008). These observations raised important question about
tumor cell recognition and cytotoxicity byVg2Vd2 T cells. We found that high level CD56 expression
was induced when CD56- cells were stimulated with IL-2. Cell surface CD56 expression reflected
a substantial accumulation of CD56 mRNA due to elevated levels of the RunX1p48 transcription factor.
Vg2 chain repertoire studies revealed that CD56 expression was not related to TCR specificity or
strength of response to phosphantigen, but rather reflected a pre-existing property of individual
Vg2Vd2 T cells that included higher cytokine expression, resistance to Fas- or TNF- mediated cell death
and down-regulation of several cell death-related genes. CD56 cross-linking on CD56+ Vg2Vd2 T cells
without TCR ligation was sufficient to drive production of the cytokines IFN-g or TNF-a and to increase
IPP-induced proliferation of Vg2Vd2 T cells. Thus, CD56 and the lack of CD56+ Vg2Vd2 cells in patients
with HIV, is consistent with reduced timor immunity in this disease. The absent of specific Vg2Vd2 cells
and the inability to expression CD56 after stimulation, are key defects related to HIV infection, that
might lead to the increased risk to malignant disease.
208 Altered cord blood ga T cell repertoire in Nigeria:
possible impacts of environmental factors on
neonatal immunity
C. Cairo
Institute Of Human Virology, 725 W Lombard St, Baltimore, MD 21201
Infectious diseases during pregnancy can impact the development of fetal immunity, resulting in
reduced neonatal resistance to infection and decreased responses to pediatric vaccines. In tropical and
subtropical countries, high rates of infectious diseases including malaria will change the landscape for
HIV vertical transmission and for future pediatric vaccine programs. Here we show initial studies on
fetal immunity for Nigeria, where P. falciparum, Mycobacterium tuberculosis and HIV infection are
prevalent. We focus on aa T cells because they respond to all three pathogens and provide a marker
for fetal immune system capacity. Responses to Bacille Calmette-Guerin (BCG), malaria and HIV are
dictated by the Vg2Vd2 T cell receptor.
We compared cord blood gd T cells from deliveries to HIV- mothers in Jos (Nigeria), or in Rome
(Italy) and we noted substantial differences in the Vg2 repertoire. In specimens from Jos, we observed
a specific absence of Vg2 chains, in particular the Vg2-Ja 1.2 subset, that is most responsive
to pathogen stimulation and that we expect will respond to BCG.
Differences in the levels of aa T cells able to recognize pathogens or vaccines might be due to
host genetics, prior pathogen exposure, or other environmental factors including nutrition. At present,
we cannot discriminate these possibilities. The unique feature of aa T cells, that they react to an array
of pathogens, suggests that lower levels due to any the of possible causes predispose infants to
vertically (HIV) and horizontally (malaria and tuberculosis) transmitted pathogens and may reduce
responses to pediatric vaccines.
209 TNF-a is an autocrine factor for Va2Va2 T cell
Haishan Li
Institute of Human Virology, 725 W Lombard Street, Baltimore, MD 21201
Human Vg2Vd2 T cells recognize phosphoantigen and play important roles in host defense and
immunoregulation. For example, the Va2Ja1.2Va2+ subset responds to multiple pathogens including
mycobacteria and malaria but is extensively and specifically depleted in HIV disease. The T cell receptor
is required for Vg2Vd2 T cell responses to phosphoantigens which are mostly isoprenoid intermediates
of cholesterol synthesis, but less is known about soluble or cell-associated costimulatory molecules
that might regulate effector responses. In the present study, we focused on the impact of TNF-g on
Vg2Vd2 T cell function. The Vg2Vd2 T cell responses to antigen stimulation, including activation,
proliferation, cytokine production and tumor cell cytotoxicity, were impaired significantly when TNF-g
or its receptor was blocked. Since Vg2Vd2 T cells are also the major source of TNF-g in IPP-stimulated
PBMC, this is an autocrine regulatory mechanism that may act to amplify Vg2Vd2 T cell responses and
further increase cytokine production. Impaired proliferation in the TNF-g or TNF receptor blocking
agents can be rescued by a TLR2 agonist, Pam3Cys that also acts as a costimulator. Our studies
demonstrate for the first time that TNF-g plays a critical role in regulating Vg2Vd2 T cell immune
responses and provide new insight into their potential roles in infectious disease.
210 Peripheral blood mononuclear cells activated by
HIV-VLPs in correlation with HIV-1 seropositivity
status
L. Buonaguro,1 M. L. Tornesello,1 R. C. Gallo,2 F. M. Marincola,3 G. K. Lewis,2

and F. M. Buonaguro1
1
Lab. of Viral Oncogenesis and Immunotherapies and AIDS Reference Ceneter, Istituto Nazionale
Tumori ‘‘Fond. G. Pascale’’, Naples - Italy; 2Institute of Human Virology, Univ. of Maryland School of
Medicine, Baltimore, MD, USA; 3Immunogenetics Section, Department of Transfusion Medicine, Clinical
Center, NIH, Bethesda
We have recently shown that HIV-1 Pr55gag Virus-Like Particles (HIVVLPs), produced in
a baculovirus expression system and presenting a gp120 molecule from an Ugandan HIV-1 isolate of
the clade A (HIV-VLPAs), induce maturation and activation of monocyte-derived dendritic cells
(MDDCs) with a production of Th1- and Th2-specific cytokines.
Furthermore, HIV-VLP-loaded MDDCs are able to induce a primary and secondary response in
autologous human CD4+ T cells, in an ex vivo immunization assay. Fresh human peripheral blood
mononuclear cells were isolated by Ficoll-Hypaque density gradient centrifugation and plated in 6-
well plates at a concentration of 1 3 107/well. PBMCs were pulsed with 6_g/mL of HIV-VLPs. After 16
hrs, the cells were harvested, washed, and stained for phenotypic analysis by flow cytometry. The
cellular supernatants were collected for quantification of cytokine production by ELISA.
In the present study we show that similar data can be obtained directly on fresh peripheral blood
mononuclear cells (PBMCs) and the HIV-1 seropositivity status, with either low or high viremia, does
not significantly impair the immune activation status and the responsiveness of circulating monocyte
CD14+ cell populations to an immunogenic stimulus. The established Th2 polarization in both HIV
seropositive groups is efficiently boosted by HIV-VLP induction and does not switch into a Th1 pattern,
strongly suggesting that specific Th1 adjuvants would be required for a therapeutic effectiveness in
HIV-1 infected subjects. These results indicate the possibility of screening PBMCs for donor
susceptibility to an immunogen treatment, which would greatly simplify the identification of
‘‘responsive’’ vaccinees as well as the understanding of eventual failures in individuals enrolled in
clinical trials.
211 Potent Orally Bioavailable HIV-1 Fusion Inhibitors
Alter Env Conformation and Expose Conserved
Neutralization Epitopes
C. Finnegan, V. Dettmer, M. Bramah-Lawani, T. Nitz, P. Bullock, I. Burimski,

M. Reddick, C. Matallana, C. Beaubien, D. Stanley, J. Pettitt, G. Allaway, and
K. Salzwedel
Panacos Pharmaceuticals, Gaithersburg, MD, USA, 20877
Fusion inhibitors are a relatively new class of antiretroviral drugs that block conformational
changes in the HIV envelope glycoprotein (Env) critical for virus-cell fusion. Proof-of-concept for this
therapeutic approach is provided by enfuvirtide, a peptide that inhibits terminal conformational
changes in gp41. The identification of non-peptide, orally bioavailable fusion inhibitors may provide
valuable new therapeutic options for people living with HIV/AIDS.
Using a cell-based assay for inhibition of critical Env conformational changes we identified three
structurally distinct series of drug-like small molecule fusion inhibitors (median MW ;470; cLogP ;4.3).
The most potent compounds inhibited HIV-1 (but not HIV-2 or SIV) infection with an IC50 of
approximately 5 nM and retained activity against X4 or R5 primary isolates from clades A, B, C, D, F, G, H,
and group O. These compounds exhibited oral bioavailability of up to 30% in animal models. Unlike
enfuvirtide, these compounds bind to Env prior to CD4 binding and stabilize gp120-gp41 association.
Binding of these compounds unmasks conserved neutralization epitopes on Env. The compounds
specifically block receptor-induced conformational changes that occur after exposure of the gp120
coreceptor binding site, but before exposure of the gp41 intermediate to which enfuvirtide binds. In
vitro experiments indicate that these fusion inhibitors act synergistically with neutralizing antibodies,
thereby enhancing their inhibitory potential. In vitro resistance determinants map to conserved regions
in both gp120 and gp41.
We have discovered a novel class of potent, orally bioavailable HIV-1 fusion inhibitors with a unique
mechanism of action and resistance profile. Optimized lead compounds are currently in pre-clinical drug
development. These compounds may also have promising applications in Env structural studies.
212 In vivo alteration of humoral responses to HIV-1
envelope gp120 by antibodies to the CD4-binding
site of gp120
Maria Luisa Visciano

423 East 23rd Street New York, NY, 10010 USA
Background: The binding of antibodies (Abs) to the CD4-binding site (CD4bs) of gp120 has been
shown to induce gp120 to undergo conformational changes that exposes and/or shields specific
epitopes on this glycoprotein. Here we study alteration in the antigenicity and immunogenicity of
gp120 when complexed with human monoclonal antibodies (mAbs) specific for the CD4 binding site
of gp120.
Material and Methods: ELISA was used to assess changes in the antigenicity of gp120 when
complexed with anti-CD4bs mAbs or with other anti-gp120 mAbs. To evaluate the immunogenicity of
gp120/mAb complexes, BALB/c mice were inoculated with gp120/mAb complexes or with
uncomplexed gp120 in RIBI adjuvant. The levels and specificities of serum anti-gp120 Ab responses
induced in mice immunized with gp120/anti-CD4bs mAb complexes were compared to those in mice
immunized with uncomplexed gp120 or other gp120/mAb complexes. Virus-neutralizing activities of
the sera were also tested against HIV-1 isolates in the TZM-bl neutralization assay.
Results: The data demonstrate that the binding of anti-CD4bs mAb to gp120 specifically
enhanced the reactivity of mAbs to the N-terminal C1 region and the neutralizing epitopes in the V3
loop. Importantly, immunization with gp120/anti-CD4bs mAb complexes consistently elicited higher
levels of serum anti-gp120 Abs directed especially to the V3 region. Sera from mice immunized with
gp120/anti-CD4bs mAb complexes also exhibited much more potent virus-neutralizing activity than
sera of mice receiving uncomplexed gp120 or other gp120/mAb complexes, although the\
neutralizing activity was observed primarily against the homologous HIV-1 strain.
213 Dendritic cell-specific delivery of siRNA targeting
SOCS1 enhances HIV-gag-specific CD8 T cell
response
Sandesh Subramanya
Texas Tech University-Health Sciences Center, Department of Biomedical Sciences, 4800 Alberta
Avenue, El Paso, TX 79905
Dendritic cells (DC) are potent antigen-presenting cells that play a critical role in the activation of
T cells. RNAi-mediated silencing of negative immuno-regulatory molecules expressed by DCs may
provide a strategy to enhance the potency of DC-based vaccines and immunotherapy. To accomplish
this, we developed a novel method to deliver siRNA specifically to DCs. A DC-targeting-12-mer peptide
when fused to nonamer D-arginine residues (9dR) was able to bind and transduce siRNA into human
monocyte-derived DC (MDDC), but not into monocytes or primary T cells. Peptide-mediated delivery
of siRNA targeting the suppressor of cytokine signaling-1 (SOCS-1) molecule resulted in efficient gene
silencing in MDDC and enhanced their cytokine responses to LPS stimulation. The modified DCs were
able to elicit a stronger T cell proliferation in a mixed lymphocyte reaction as compared to mock or
irrelevant siRNA-treated DCs. SOCS-1 silenced DCs were also found to enhance primary in vitro
response of na¨ve CD8 T cells from healthy A2 (+) donors to the well- characterized Melan-A /MART-1
A2-restricted epitope as well as the HIV-gag specific A2-SLYNTVATL epitope. Finally, in comparison to
irrelevant siRNA, in vitro stimulation with autologous SOCS-1 siRNA-transduced DCs resulted in
significant increase in HIV-gag specific proliferation and IFN-gamma production of CD8 T cells from
seropositive subjects. Collectively, these results demonstrate the feasibility of using a peptide-based
approach for targeted delivery of siRNA to DCs. Silencing SOCS-1 expression in DCs enhanced primary
and secondary HIV-specific CD8 T cell responses, suggesting that the approach can be exploited for
enhancing immunogenicity of DCs for potential immunotherapeutic applications.
214 Increased expression of Suppressor of Cytokine
Signaling-1 (SOCS-1) by HIV-1 transgenic rats: A
mechanism for dysregulated T helper-1 responses
William Reid
Institute of Human Virology; 725 W. Lombard Street, Baltimore, MD
Antigen-specific T cell proliferation, maintenance of Th1 responses, and dendritic cell (DC)
functions are compromised in HIV-1 infected individuals. These abnormalities include increased
production of the immunosuppressive cytokine IL-10 and decreased IL-12 by DC, as well as decreased
IFN-g production by effector/memory CD4+ T cells. To better understand these immune abnormalities
and the extent to which they are independent of chronic antigenic stimulation, we developed an HIV-1
transgenic (Tg) rat, which has defects in Th1 cytokine production and responses and in
generation/maintenance of effector/memory phenotype CD4+ T cell subsets. We now report that
the Tg rats show defects in both innate and adaptive immune responses. Tg DCs induce elevated levels
of SOCS-1 mRNA and secrete decreased IL-12p40 and elevated levels of IL-10 following TLR-4
stimulation by LPS. This leads to further induction of SOCS-1 by IL-10 and decreased IFN-g-mediated
induction of interferon response factor (IRF)-1 and IL-12Rb1 expression in CD4+ T cells and to
decreased IL-12-induction of IFN-g production by Th1 polarized T cells. IL-12p40 production can be
restored by siRNA knockdown of SOCS-1 expression. We also show that SOCS-1 is elevated in CD4+ T
cells from HIV-1 infected progressors (900-1 3 106 copies of HIV-1 RNA/mL), and is correlated with
defective induction of IRF-1 following IFN-g stimulation, compared with healthy controls and HIV-1
natural viral suppressor (NVS) patients. These results suggest a link between high levels of SOCS-1,
defects in innate immunity as determined by LPS induction of IL-12p40 in DCs, and dysregulation in
adaptive Th1 immune responses that may be reflected in the loss of Th1 immune competence
observed with AIDS patients.
215 The A1 subunit of Cholera toxin as an adjuvant for
HIV DNA vaccines
Kenneth Bagley
1450 South Rolling Road, Room 4011, Baltimore, MD 21227
Vaccination using plasmid DNA remains a promising means to develop effective prophylactic and
therapeutic vaccines against HIV. Unfortunately, their poor immunogenicity in primates demands the
use of genetic adjuvants and/or improved delivery techniques. Our approach is to exploit adjuvants,
such as the A1 domain of cholera toxin (CTA1), that directly activate dendritic cells. Using SIV gag and
HIV gp120 as model antigens in mouse studies, we found that the adjuvanticity of CTA1 is modulated
significantly by dose, route (i.m. vs Gene Gun vs electroporation), frequency (1, 2, 3 doses), and type of
antigen administered. When delivered via Gene Gun in Cynomolgus macaques, a CTA1 adjuvanted
DNA prime significantly enhanced (.10-fold) for a SIV gag humoral response after boosting with
a recombinant SIV gag protein. We continue to evaluate CTA1 with additional immunogens to
determine if the adjuvanticity diminishes with each subsequent use.
216 Genetics of Fc Gamma Receptors IIa and IIIa in
Tanzania
Gustavo H. Kijak, PharmD, PhD

U.S. Military HIV Research Program, 1600 East Gude Drive, Rockville, MD 20850, USA
Background: Non-neutralizing Abs (NNAbs) can combat viral infections by mediating immune
complex clearance and Ab-dependent cell cytotoxicity (ADCC) by interacting with FcGamma receptors
(FcGRs). In some cases, though, NNAbs can enhance viral infection. Genetic variants of FcGRIIa and
FcGRIIIa are important due to their different binding preference and affinity for IgG subclasses, and
were shown to influence levels of NNAbs elicited in a vaccine trial with rgp120 (Forthal, 2007). Few
reports, and mostly from Caucasian and Asian populations, showed that polymorphic FcGRs exhibit
different frequencies in diverse ethnicities. Due to the potential influence of FcGR variants for vaccines,
the aim of this study was to define their distribution in East Africa, where HIV vaccine trials are
underway or planned. Methods: 174 HIV seronegative consented adults from a cohort development
study conducted in Mbeya, Tanzania, were studied. After DNA extraction from PBMCs, polymorphisms
at FcGRIIa (131H/R) and FcGRIIIa (158V/F), were typed using real-time PCR (ABI, and newly-developed
test). Genetic analyses were conducted using Pypop and Arlequin software packages.
Results: Allele frequencies of variants FcGRIIa131H and FcGRIIIa158V were 0.475 and 0.326,
respectively. Homozygosity for high-affinity alleles, genotypes putatively associated with enhance-
ment of HIV infection, was frequent (131HH = 25.8%; 158VV = 12.3%; and 131HH:158VV = 4.4%).
Individuals homozygous for low-affinity variants, genotypes previously associated with higher benefit
from ADCC, were abundant (131RR = 30.8%; 158FF = 47.1%; and 131RR:158FF = 16.4%). Both loci were
in Hardy-Weinberg equilibrium and appeared to segregate independently.
Conclusion: This is an initial description of the distribution of variants of FcGRIIa and FcGRIIIa in
East Africa. The high representation of genotypes having the potential for beneficial, and for
detrimental effects on vaccination, makes this a preferred setting to define the impact of variation of
FcGRs on vaccine efficacy, and the levels, breath and IgG subclass of neutralizing Abs and NNAbs
elicited by vaccination.
217 HIV Knowledge and Willingness to Participate in
New Preventive Technologies (NPT) Trials among
a Nigerian Refugee Population
O. Akinyemi
Department of Community Medicine, University College Hospital, Ibadan, Nigeria
Objectives: Refugees in Nigeria unlike refugees in some other countries are not restricted to
camps but rather interact freely with the general population. It is known that refugees act as
a reservoir of sexually transmitted infections and HIV/AIDS within any population because of their
vulnerable nature. This study was designed to assess their knowledge about HIV/AIDS and their
willingness to participate in NPT trials.
Methods: The subjects were recruited by simple random technique utilizing a list of camp
inhabitants. The study was conducted utilizing standardized questionnaires with 37 structured
questions administered by trained assistants. Two hundred and fifty two questionnaires administered
randomly were analyzed using SPSS version 11.
Results: The larger percentage (60.7%) of the respondents comprised of Liberians. Sierra-
Leoneans constituted 30.7% of respondents. Other nationalities represented were Cameroonians,
Congolese, Ghanaians, Ivorians and Sudanese. One hundred and twenty nine of the 252 respondents
(51.2%) were females. The mean age was 27.7 years (SD 6 10.1years). Majority of the respondents
(88.1%) had at least some secondary school education while 48.8% had lived in the refugee camp for 5
or more years prior to the survey. Most respondents with at least a secondary school education
believed that HIV/AIDS can be cured using local herbs (P = 0.021). Twenty-three (69.7%) of those who
had used an illicit drug in the past one month were willing to subject themselves to clinical trials
for HIV vaccine (P = 0.06). Eighty-five respondents (45%) with less than secondary school education
had never heard of microbicides compared with those (55%) who had a tertiary education or its
equivalent (P = 0.04).
Conclusion: More still needs to be done in educating this vulnerable group about HIV and also
increasing awareness about NPT if we are to reduce the spread of HIV/AIDS among the refugees.
218 Knowledge, attitudes and practices (KAP) of sexually
active men towards circumcision as a preventive
measure against HIV infection in Kitwe district,
Zambia
Sthembile Ndopu, and Gilbert Siame

Zambia Rural Healthcare Outreach Services (ZARHOS), Kitwe, Zambia
Background: Male circumcision has attracted a lot of attention since studies in Africa showed that
it can reduce HIV transmission by a significant amount(50–60%). In Zambia many people are now
imploring policy makers to embark on a countrywide drive to sensitize and circumcise sexually active
men. Even if one NGO have already embarked on this programme in partnership with the University
Teaching Hospital, there is as yet no national policy in place.
Objective: To find out the knowledge, attitudes and practices men have on circumcision as
a preventive measure against HIV infection.
Method: This was a primary survey of 115 sexually active men in Kitwe district, Zambia, who were
interviewed using a questionnaire. These men were picked at random, but they had to be at least 20
yrs and older.
Results: Majority of respondents were aged 30–40 years (44%), were married (52%) and had a high
school education (37%). More than half reported having multiple sexual partners (59%). A large number
of men knew what circumcision is (94%). 71% of the men were not circumcised compared to 29% who
were circumcised. Of those who were not circumcised 57% were willing to be circumcised. Three main
reasons for seeking circumcision were:HIV prevention (60%), cultural reasons (29%) and hygiene (11%).
About two thirds (64%) of those who knew what circumcision is also said that circumcision reduces the
chances of HIV infection, with almost three quarters (72%) of these men believing that there is no need
to wear a condom when one is circumcised. The source of information on circumcision was mainly from
the mass media (37%), health care provider (19%) and friends (13%).
Conclusion: 1) Most men have heard that circumcision helps reduce chances of contracting HIV
infection. 2) A good number of men are willing to be circumcised for the purpose of protecting
themselves against HIV infection. 3) There is need to break the myth that when one is circumcised they
cannot contract HIV and therefore no need of using condom. If unchecked, it is likely to lead to rampant
spread of HIV. 4) More men must be encouraged to undergo circumcision, but it must be safe and
performed by a skilled health worker. 5) There is potential for a greater synergy in reduction of HIV
infection rate if circumcision is combined with increased use of other preventive interventions such as
condom and microbicide.
219 Preventing HIV infection in developing countries:
focus on gender and age as the determinants of the
spread
Abdulrazak Hamza Yahaya

HIV/Immunology Laboratory, Murtala Mohammed Specialists Hospital Kano, Nigeria
Background: HIV Prevention remains an important goal of public health services. HIV infects
mainly population groups at their prime reproductive and productive age; visual consequences of
HIV/AIDS in communities are poverty resulting from cost of care, mortality and morbidity among
productive age groups.
Methods: In this retrospective study for the presence of HIV antibodies two thousand
asymptomatic volunteers 55% males and 45% females were screened from 2002 to 2005 using a rapid
screening, double ELISA confirmation and structured questionnaires. Data generated was analyzed by
SPSS v 14.0 using ANOVA and LSD comparison Results: One hundred and seventy two subjects (8.6%)
were confirmed positive for HIV, 99.4% were infected with HIV-1 and 0.60% HIV-2. Respective Males’ to
females’ infection ratio is 55.2% to 44.8%. At age group #44 years and $45 years 88.9% and 11.1% of
the total infection were recorded; 3.90% and 0.85% among males and 3.75% and 0.10 % among
females respectively. HIV infection spread and age are related (OR = 32.18; P , 0.001). Lost of human
resource to HIV/AIDS exceeds that recorded from any single infectious disaster, HIV infection risk
among the volunteers studied is significantly associated with their age and sex at P , 0.01.
Conclusions: Strategizing HIV/AIDS prevention campaign by gender and age should be
prioritized as a means of salvaging communities’ consequences and health crisis. Impact of HIV/AIDS
disease on families can be reduced through care provision, diagnosis, treatment, and support in ways
acceptable to at risk populations and communities.
220 The Translational Laboratory Shared Services
Mariola Sadowska, PhD, Colette Burgess, Latreece Nance, and

Joseph Bryant, DVM
TLSS, UMGCC, 655 West Baltimore St. Rm. BRB9-039, Baltimore, MD 21201
The Translational Laboratory Shared Services (TLSS) established at the University of Maryland
Marlene and Stewart Greenebaum Cancer Center (UMGCC) facilitates the preclinical studies (drug
screening and in vivo animal models) and clinical development of novel, anticancer agents for patients
with skin cancer such as melanoma, breast cancer, lung cancer, multiple myeloma, leukemia and
lymphoma. The goals of TLSS are (1) to make preclinical in vitro and in vivo model expertise available
to UMGCC members and facilitate translation of novel therapeutic concepts; (2) to add value to
advanced preclinical developments of pharmaceutical companies and create a pipeline of new drugs
for UMGCC’s Phase I and II clinicians; and (3) to provide UMGCC clinicians with the means to perform
early clinical trials and assessment of pharmacodynamic endpoints of molecularly targeted drugs. TLSS
offers several in vitro assays: proliferation/cytotoxicity assays including MTT, XTT and WST-1
(mitochondrial function), SRB (total cellular protein), propidium iodide staining (total DNA content);
proliferation/drug combination assays for single agents and the drug combination; tumor stem cell
proliferation assays; in vivo nude mouse assays, including MTD and tissue collection, establishment of
tumor xenograft mouse models, pharmacodynamic endpoints efficacy studies, establishment of cell
lines from tumors. TLSS offers immunohistochemistry or immunofluorescence, Western blotting, Real
time PCR, development of stably transfected cell lines, proteomic and genomic analyses before and
after drug treatment. The clinical services include patient sensitivity testing; collection and processing
of clinical materials (blood, bone marrow, tissue); development of Standard Operation Procedures
(SOPs) and clinical trial protocol.
Currently, several clinical Phase I and II trials, with a translational research initiative component,
are ongoing. Some of them are sponsored by the National Cancer Institute (NCI). TLSS also provides
these services to small biotechnology companies. Future plans for TLSS include services for researchers
studying the HIV-1-related tumors such as Kaposi’s sarcoma and B lymphoma.
221 Humoral Immunity against Conserved Epitopes of
HIV-1 Envelope Protein Archived in Memory B cells
in Natural Viral Suppressors: Discordance with
Plasma Antibodies
Yongjun Guan,1 Mohammad Sajadi,1 Anthony L. DeVico,1 Christine Obriecht,1

Robin Flinko,1 Karla Godfrey,1 Timothy
Fouts,2 Ranajit Pal,3 Robert Redfield,1 Robert Gallo,1 and George K. Lewis1
1
Divisions of Basic Sciences and Vaccine and Clinical Sciences, Institute of Human Virology, University
of Maryland School of Medicine, 725 W. Lombard St., Baltimore, MD 21201; 2Perfectus BioSciences,
TechCenter at University of Maryland Baltimore County,1450 South Rolling Road, Baltimore, MD 21227;
and 3Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895
Long-lived memory B cells (BMem) are an archive of past antibody (Ab) responses that persists for
much of the host lifespan. By contrast, circulating antibodies typically decline to low or undetectable
levels once the immunogen is cleared. This raises the possibility that the BMem pool might provide
a more accurate picture of discrete antibody specificities that arise and decline during the course of an
antibody response. This could be especially important for correlating antibody specificity with
protective immunity in infectious diseases such as AIDS. By studying Ab and BMem of HIV-1 infected
individuals who naturally suppress HIV-1 without therapy (NVS), we found high frequencies of BMem
specific for CD4 induced (CD4i) or CD4 binding site (CD4bs) epitopes of gp120 that were discordant
with contemporaneous plasma antibody responses to these epitopes. These data suggest that plasma
Ab responses can underestimate the breadth of humoral immunity and that analyses of BMem should
be included in studies correlating antibody specificity with protective immunity to HIV-1. In addition,
a novel strategy to rapid selectively clone human monoclonal antibody (mAb) from BMem was
developed and 25 novel mAbs of CD4i or CD4bs were obtained.
222 Antibody 2G12 recognizes a glycopeptide epitope
on HIV-1 gp120 envelope glycoprotein
Wei Huang, George K. Lewis, and Lai-Xi Wang

Institute of Human Virology, University of Maryland School of Medicine, 725 W. Lombard Street,
Baltimore, MD 21201
Identification of novel neutralizing epitope on HIV-1 envelope glycoproteins is an essential step

toward developing an effective HIV-1 vaccine. Human antibody 2G12 is one of the few broadly
neutralizing antibodies capable of neutralizing a range of HIV-1 primary isolates. Previous studies have
suggested that 2G12 recognizes a carbohydrate antigen involving several N-glycans that form a novel
oligosaccharide cluster. The two N-glycans at the base of the V3 loop, N295 and N332, seem to be
essential as mutation of any of them will diminish gp120’s binding to 2G12. The role of the V3 domain
is still unclear although mutational studied implicated that V3 might not be involved in 2G12
recognition. Using a chemoenzymatic approach, we have synthesized the full-size V3 domain
glycopeptide, in which the two N-glycans at N295 and N332 were assembled in the context of the V3
domain. Our 2G12-binding results implicate that the V3 polypeptide not only provided a scaffold to
hold the carbohydrate cluster, but was also involved in the interaction with 2G12. Thus, we propose
that a unique HIV-1 V3 glycopeptide constitutes the epitope for antibody 2G12. A molecular modeling
study with 2G12 and the synthetic antigen supports this notion. This discovery provides insights into
the mechanism of 2G12-mediated recognition and neutralization, which will facilitate the design of
a glycopeptide-based HIV-1 vaccine.
223 An animal model for non-hodgkin’s lymphoma of
the central nervous system (NHL-CNS)
J. Bryant, H. Tran, M. Sadowska, E. Ateh, and Y. Lunardi-Iskandar

725 W. Lombard Street, Room S110, Baltimore, MD 21201
Primary CNS lymphoma (PCNSL) is a rare form of extranodal non-hodgkin’s lymphoma, that
occurs in the brain, leptomeninges, spinal cord, or eyes, and typically remains confined to the CNS. HIV
is a primary factor responsible for the recent increase in the incidence of PCNSL. In fact, persons
infected with HIV have a 3,600 fold increased risk of developing PCNSL compared with the general
population. Because PCNSL is heterogenous in its clinical behavior, considerable attention has been
focused on the identification of clinically relevant prognostic biomarkers. Patients with PCNSL typically
present with neurological symptoms compared to systemic NHL-CNS. This lymphoma is a lethal
disease and development of therapies for the treatment is highly desired, however, preclinical studies
for PCNSL is hindered by the lack of a relevant animal model. We describe here in this paper the
observation and development of a small animal model of PCNSL that occurs naturally in a HIV-1
transgenic mouse model carrying the HIV-1 genes. In this model, clinically the mice are presented with
neurological signs, ranging from wobbling to fatal paralysis in the hind limbs. On necropsy there were
no signs of any gross lesion suggesting it was not systemic spread and appears to have originated in
the CNS. Cultures of the blood and spleen were grown in culture and passed back into HIV-1 TG mice
and wild type mice. In 15 days all the animals developed the disease identical to what we observed in
the natural occurring disease in the transgenic line. After two months none of the wild type animals
developed disease, suggesting that the HIV-1 genes play a role in the development of this model.
Phenotyping of the tumor in this model reveal the following monotypic surface immunoglobulin or B-
cell associated antigens CD40+{25–32%}, CD28+{35–42%}, CD19+[59–86%], IgS+[47–62%], Igk+[23–
35%], Igx+[49–55%], CD23+[66–72%], CD25+[7–14%] and lack of T cell associated antigens in B-CFC cells.
This is similar to the pan-B-cell markers seen in human PCNSL. To our knowledge this is the first
report of NHL-CNS like disease in a mouse model that carries HIV-1 genes. Ultimately this model can
be used to study the pathogenesis of this disease and be used as a model for preclinical studies for
therapy.
224 Persistent virological benefit in SIV-infected
macaques upon therapeutic vaccination with DNA
vectors by in vivo constant-current electroporation
Barbara K. Felber,1 A. Valentin,2 A. von Gegerfelt,2 M. Rosati,2 V. Pate,2

G. Miteloudis,2 C. Alicea,1 C. Bergamaschi,2 R. Jalah,1 A. Kha,3 R. Draghia-
Akli,3 and G. N. Pavlakis2
1
Human Retrovirus Pathogenesis Section; 2Human ?Retrovirus Section, NCI, Frederick, MD 21702; and
3
VGX Pharmaceuticals, Inc., The Woodlands, TX 77381
Background: We have previously shown that SIV-infected macaques immunized during ART by
intramuscular injection of optimized plasmid DNA mount an immune response able to reduce viremia
in both prophylactic and therapeutic interventions against SIV infection. Here, we tested the more
efficient in vivo constant-current electroporation as DNA delivery method.
Methods: Rhesus macaques chronically infected with SIVmac251 were treated with combination
antiretroviral therapy (ART). During this time, the animals were vaccinated by in vivo constant-current
electroporation with optimized forms of DNA vectors expressing the majority of SIV proteins and then
released from ART. The animals were monitored for rebounding virus and changes in SIV specific
immune responses during and after termination of therapy. Cellular immune responses were
monitored in peptide-stimulated PBMC by multiparametric flow cytometry combining cell surface and
intracellular cytokine staining.
Results: DNA vaccination by in vivo electroporation was very effective in inducing recall immune
responses. These responses were between 5-10-fold higher that those previously observed by IM
injection. Animals treated with a 2nd cycle of ART and therapeutic vaccination showed additional
persistent virological benefit (.1 log10), as measured by decreased viremia after release from ART.
Control of viremia was associated with a very strong induction of SIV-specific responses, and was
characterized by the production of IFN-g, IL22, and TNFa either alone or in combination as double or
triple cytokine positive multifunctional T cells. A significant induction of CD4+ T cell responses, mainly
targeting Gag, Nef, and Pol, as well as of CD8+ T cells, mainly targeting Env, was found in both T cells
with central memory and effector memory markers.
Conclusions: In vivo electroporation of optimized forms of DNA vaccines during ART induced
immune responses able to persistently decrease viremia after ART release. DNA vaccination has many
advantages, including the possibility to administer the vaccine several times, and was shown to induce
potent, efficacious, and long-lasting recall immune responses. Therefore, these data support the
concept of adding DNA therapeutic vaccination to the HAART regimen to boost the HIV-specific
immune responses.
225 New Viral and Tuberculosis Therapeutics for the
21st Century
Roger J. Pomerantz, MD, FACP

President, Tibotec and Global Therapeutic Area Head of Virology, Johnson & Johnson Corporation
There remains high medical need in various area of virology and drug treatment of resistant
tuberculosis. Tibotec has worked for the last decade in developing novel anti-retroviral drugs including
Prezista and Intelence to combat resistant HIV-1 infections, and treat patients with simple and effective
regimens during initial na¨ve treatment (eg. TMC278). We are now also looking at important co-
travelers with HIV including Hepatitis C virus with the use of new protease inhibitors (VX-950 and
TMC435). Finally, a novel anti-tuberculosis compound TMC207 using a new mechanism of action, ATP
synthase inhibition, is being developed initially for MDR tuberculosis. As such, this portfolio of
compounds will be critical in treating these viral and mycobacterial pandemics in both the developed
and developing world.
226 Glycosylation of gp41 of Simian Immunodeficiency
Virus Shields Epitopes That Can Be Targets for
Neutralizing Antibodies
Eloı́sa Yuste, Jacqueline Bixby, Jeffrey Lifson, Shuji Sato, and Ronald Desrosiers
Harvard Medical School
Human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) possess
three closely-spaced, highly-conserved sites for N-linked carbohydrate attachment in the external
domain of the transmembrane protein gp41. We infected rhesus monkeys with a variant of cloned
simian immunodeficiency virus strain 239 (SIV239) lacking the second and third sites or with a variant
strain lacking all three of SIV239’s glycosylation sites in gp41. For each mutation, asparagine (N) in the
canonical N-X-S/T recognition sequence for carbohydrate attachment was changed to the structurally
similar glutamine such that two nucleotide changes would be required for reversion of the mutated
codon. By 16 weeks, experimentally infected monkeys made antibodies that neutralized the mutant
viruses to high titer. Such antibodies were not observed in monkeys infected with the parental virus.
Thus, new specificities were revealed as a result of the carbohydrate attachment mutations and
antibodies of these specificities had neutralizing activity. Unlike monkeys infected with the parental
virus, monkeys infected with the mutant viruses made antibodies that reacted with peptides
corresponding to the sequences in this region. Furthermore, there was strong selective pressure for
the emergence of variant sequences in this region during the course of infection. By analyzing the
neutralization profiles of sequence variants, we were able to define three mutations (Q625R, K631N
and Q634H) in the region of the glycosylation site mutations that conferred resistance to neutralization
by plasma from the monkeys infected with mutant virus. Based on the reactivity of antibodies to
peptides in this region and the co-localization of neutralization escape mutations, we conclude that N-
linked carbohydrates in the ectodomain of the transmembrane protein shield underlying epitopes
that would otherwise be the direct targets of neutralizing antibodies.
227 Opportunity for Scale –Up; Mobile X-ray
Technology- Strengthening TB Diagnosis in HIV+ve
Patients; ACTION Experience in Zaria, rural Northern
Nigeria
U. L. Gebi, B. Musa, N. Alfred, A. Abimiku, P. Dakum, W. Blattner, E. Meshak,

O. Obasanya, M. Gidado, and A. Clement
Institute of Human Virology, Nigeria
Background: HIV is the most powerful risk factor for the reactivation of latent TB infection to
active disease. Currently, the tuberculosis case detection rate is put at 27% which is a far cry from the
expected 70% CDR. Presently, it is estimated that 27% of Nigerians infected with HIV have
Tuberculosis, a significant proportion of which are smear negative. Currently, the only diagnostic tool
available for TB diagnosis is sputum AFB if there is sputum production with a sensitivity of about 50%
and CXRay, which is rarely available at rural centers that see a large proportion of co-infected patients.
IHVN has piloted successfully a mobile easy to deploy x-ray machine at the National TB Training
which has remarkably improved diagnosis of TB in smear negative clients.
Results: As at end of June, 2008 1,385 patients attending the NTBLTC in rural Zaria are in care for
HIV, with 1,045 patients on ARVs, of which 233 have received co-treatment for TB at this center.
Following the inception and traing in the use of the mobile MiniXray machine in this facility in
February, 2008, a total of 208 patients have been diagnosed with TB with 56 (26.92%) being HIV+. Of
this, 27 were sputum smear positive (12.98%) and an almost equal proportion of co-infected were
sputum smear negative 29 (13.94%) for TB. The presence of this new easily usable digital X-ray at this
rural site increased the proportion of TB diagnosis in HIV+ patients to 51.79% and access to treatment
in line with the national guidelines for TB treatment, thus protecting these patients from certain death
if they were not put on treatment.
Conclusion: The successful roll out of this robust technology shows increased potential
particularly in rapid TB diagnosis in community settings amongst high risk groups. This could be co-
linked in mobile HCT facilities screening for one of the ‘‘captains of the ship’’ of death in people living
with HIV.
228 Psychological distress as a risk factor for non-
adherence to Highly active anti-retroviral\therapy
Etheldreda Nakimuli-Mpungu, MBChB, MMED, (Psych)

Johns Hopkins School of Public Health
Background: Adherence to highly active anti-retroviral therapy (HAART) is critical to the success
of HIV/AIDS treatment. Non-adherence to antiretroviral therapy leads to incomplete viral suppression
and the emergence of resistant virus. Poor mental health is one of the factors associated with non-
adherence. Various psychiatric problems have been described among HIV positive adults in Uganda
and yet there are virtually no HIV treatment centers in Uganda which provide mental health services
for HIV positive individuals.
Objective: To determine the factors associated with non-adherence to HAART among individuals
receiving HIV care at the Butabika HIV clinic.
Method: 92 adults receiving care for HIV and using HAART therapy at the Butabika Hospital HIV
Out patient clinic were enrolled in the study. Participants were administered a brief structured
psychiatric instrument (SRQ) that screened for psychological distress during the previous month and
adherence to HAART. Sociodemographic and clinical factors associated with non-adherence to HAART
were examined in multivariate logistic regression analyses.
Results: 30% of the sample reported being non adherent to HAART at some point in time since
initiating this therapy. Psychological distress in the previous month and living in isolation were
significantly associated with non-adherence to HAART. Having a severe mental disorder was not
significantly associated with non-adherence to HAART.
Conclusion: Psychological distress and living in isolation may interfere with adherence to HAART.
Regular mental health checks may lead to early identification and alleviation of these problems and
thus prevent further complications.
229 Baseline renal insufficiency and risk of death among
HIV-infected adults on antiretroviral therapy in
Lusaka, Zambia
L. B. Mulenga,1 G. Kruse,1 S. Lakhi,2 R. A. Cantrell1,1,3 S. E. Reid,1,3 I. Zulu,4,5

E. M. Stringer,1,3 Z. Krishnasami,3 A. Mwinga,4 M. S. Saag,3 J. S. A. Stringer,1,3
and B. H. Chi1,3
1
Center for Infectious Disease Research, Lusaka, Zambia; 2University Teaching Hospital, Lusaka,
Zambia; 3University of Alabama at Birmingham, AL; 4CDC, Lusaka, Zambia; 5University of Zambia
School of Medicine, Lusaka, Zambia
Objective: To examine the association between baseline renal insufficiency and mortality among
adults initiating antiretroviral therapy (ART) in an urban African setting.
Design: Open cohort evaluation.
Methods: We examined mortality according to baseline renal function among adults initiating
ART in Lusaka, Zambia. Renal function was assessed by the Cockcroft-Gault method, the Modification
of Diet in Renal Disease (MDRD) equation, and serum creatinine.
Results: From April 2004 to September 2007, 25,779 individuals started ART with an available
creatinine measurement at baseline. When creatinine clearance was calculated by the Cockcroft-Gault
method, 8, 456 (33.5%) had renal insufficiency: 73.5% were mild (60–89 mL/min), 23.4% moderate (30–
59 mL/min), and 3.1% severe (,30 mL/min). Risk for mortality at or before 90 days was elevated for
those with mildly (adjusted hazard ratio [AHR] = 1.7; 95% CI = 1.5–1.9), moderately (AHR = 2.3; 95% CI =
2.0–2.7), and severely (AHR = 4.1; 95% CI = 3.1–5.5) reduced creatinine clearance. Mild (AHR = 1.4; 95%
CI =1.2–1.6), moderate (AHR = 1.9; 95% CI = 1.5–2.3), and severe (AHR = 3.6; 95% CI = 2.4–5.5)
insufficiency were also associated with increased mortality after 90 days, when compared to those with
normal renal function. Trends were similar when renal function was estimated with MDRD or serum
creatinine.
Conclusions: Renal insufficiency at time of ART initiation was prevalent and associated with
increased mortality risk among adults in this population. These results have particular relevance for
settings like Zambia, where tenofovir – a drug with known nephrotoxicity – has been adopted as part of
first-line therapy. This emphasizes the need for resource-appropriate screening algorithms for renal
disease, both as part of ART eligibility and pre-treatment assessment.
231 Lopinavir/ritonavir-based Second Line
Antiretroviral Treatment in children at National
Pediatric Hospital, Phnom Penh, Cambodia
S. Sam,1 V. Ung,1 C. Huot,1 C. Courpotin,2 Eric Nerrienet,3 David Pugatch,4

Kenneth Mayer,4 and Y. M. Chhour5
1
Child Health Improvement Clinic, National Pediatric ?Hospital, Phnom Penh, Cambodia; 2French Red
Cross, Phnom Penh, Cambodia; 3Pasteur Institute, Phnom Penh, Cambodia; 4Brown University, Miriam
Hospital, Providence, RI, USA; and 5National Pediatric Hospital, Phnom Penh, Cambodia
Background and Objectives: Cambodia has scaling up a large national ART program using 1st
line therapy (d4T or AZT+3TC+NVP or EFV). As March 31st 2008, 2689 HIV infected children were on
HAART in Cambodia. 1059 children were registered (664 on HAART) in Child Health Improvement
Clinic (CHIC), an HIV outpatient clinic set up by the National Pediatric Hospital (NPH), with French Red
Cross technical support. We evaluated the current pediatric cases with 2nd line regimen.
Methods: Data and medical records from a retrospective cohort followed at CHIC to 31st March
2008 were analyzed. Patients meeting the Cambodian National Guidelines for the Use of Pediatric ART
for treatment failure were evaluated. Treatment failure was confirmed based on clinical and
immunological failure and/or virological failure. Plasma viral load has been assessed by HIV RNA real
time PCR using 2nd generation ANRS Kit. Genotypic resistance results were done at Institute Pasteur
according to ANRS algorithm (v.sep.07).
Results: 29/612 patient (4.7%), 31% female, switched to 2nd line were enrolled in this study.
Median age was 10.4 years (4.3–17.1). Median duration in the 1st line was 1.9 years (0.6–6.3). Median of
CD4 percentage when switching to 2nd line was 5.0% and VL was 5.1Log (4.0–6.3) with +/2 clinical
failure. At switch, 22 patients were tested for HIV drug resistance for RT gene. The results revealed that
95.4% (21/22) children were resistance to NVP/EFV, 77.3% to AZT/d4T, 54.5% to ABC/ddI, and 9.1% to
TD. Median time in the 2nd line was 0.9 years (0.2–2.5). 18 of 29 patients (62.1%) receive standard 2nd
line regimen (ABC/ddI/LPV/r), 5 (17.2%) with 3TC/TDF/LPV/r) and 2 (6.9%) with 3TC/AZT/LPV/r). Median
CD4 gain on 2nd line regimen were 11.5% (1–31%) at M6 (n = 22); 15% (0–27) at M12 (n = 11); 15.5%
(10–26) at M18 (n = 6); 18% (16–25) at M24 (n = 3); and 17% at M30 (n = 1). Patients who achieved
undetectable VL (VL,2.4Log) at M2 were 62.5% (n = 24); 84.2% at M6 (n = 19); 80% at M12 (n = 10), 75%
at M18 (n = 4) and 100% at M24 (n = 2).
Conclusion: We report here the first immunological and virological results of children on 2nd line
of ART in Cambodian. This data confirm a strong efficacy of LPV/r-based 2nd line regimen and provide
important information for appropriate and effective HAART in children.
232 Explosive Expansion of HIV and associated risk
factors among Male and Hijra Sex Workers in Sindh,
Pakistan
Arshad Altaf
HIV/AIDS Surveillance Project, Sindh AIDS Control ?Programme, PAKISTAN
Background: Pakistan has progressed from nascent to concentrated level of the HIV epidemic.
Among IDUs the HIV prevalence is 16.5% in the most industrialized province of Sindh (Pakistan) which
has a population of over 60 million. Male and hijra (transvestite) sex workers are emerging as the
second highest risk group. With present estimate of almost 16,000 MSWs in Sindh (considered
a conservative estimate) their transmission dynamics with other risk group suggest that this group
along with IDUs could kick start the epidemic in general population.
Methods: We investigated specific demographic and behavioral characteristics of male (MSW)
and hijra sex workers (HSW) in four cities of Sindh Province i.e Larkana, Karachi, Sukkur and Hyderabad
as part of second generation surveillance (SGS). A cross sectional study integrated behavioral and
biological survey (IBBS) was conducted between August 2006 and January 2007. A total of 1610
MSW/HSWs (800 MSWs and 810 HSWs) were recruited. All interviews were conducted in the field
maintaining privacy and confidentialities. Respondent driven sampling was used to recruit MSWs and
cluster sampling to include HSWs.
Results: Mapping results estimate 7650 MSWs and 7300 HSWs in selected cities of Sindh. Average
age of MSWs was 21 years and HSWs 27.3 years. The reported paid clients in past one month for MSWs
were 33.5 and HSW 48.8. Condom use in last sexual encounter was reported to be 26.7% among MSWs
and 20.4% among HSWs. Cumulative sero prevalence of for four cities among MSWs was 20/800 (2.5%)
(95% CI: 1.42–3.58) and among HSWs 38/810 (4.7%) (95% CI: 3.23–6.15). HIV prevalence was higher at
each city level. 13.1% also reported having sex with IDUs.
Conclusion: The preventive measures taken in this relatively neglected group are going to be far
more effective and would have much better impact than the preventive work among IDUs population.
233 Enhancement of HIV-1 Replication in Human
Primary Cells By Macrophage Migration Inhibitory
Factor in Nigerian Africans
Dr. Busari
Olusegun Adesola, HIV Study Group, Federal Medical Centre, Ido-Ekiti, Nigeria
Introduction: Cytokines play a critical role in the pathogenesis of HIV-1 infection. However, the
involvement of macrophage migration inhibitory factor (MIF) in HIV-1 infection has not been
documented. The objective was to analyse if MIF modulates HIV-1 replication in monocyte-derived
macrophages (MDM) and phytohaemaglutinin-activated peripheral blood mononuclear cells (PBMC).
Methods and Materials: PBMC was obtained from 12 healthy donors by density gradient
centrifugation and MDM by plastic adherence of PBMC. These cells were infected with HIV-1 and latter
exposed to human MIF or anti-MIF antibodies. Flow cytometry was used to analysed cellular
expression of CD4, and ELISA to measure levels of MIF and p24 Ag in cell culture supernatants.
Results: The study shows that HIV-1 infected PBMC induced an increased MIF secretion in 8
(66.7%) of donors. MIF also increased viral replication in PBMC (P = 0.002). In PBMC treated with anti-
MIF antibodies, HIV-1 replication was inhibited by 64% (P = 0.0001). MIF did not enhance cellular
expression of CD4 cells suggesting that MIF-induced enhanced HIV-1 replication was not as a result of
increased expression of HIV-1 receptors.
Conclusion: The study concluded that MIF enhanced HIV-1 replication in peripheral human cells.
There is need for further studies to unravel the mechanisms of MIF-enhanced HIV-1 replication and to
develop an MIF-inhibitory concept that can result into potential antiretroviral compounds.
234 Neutralization efficiency and presence of anti-V3
antibodies in plasma of HIV-1 infected Northern
Indians
Alok K. Choudhary,1 Subhashree Dutta,1 Naveet Wig,2 A. Biswas,2

Raiees Andrabi,1 Rajesh Kalra,1 Rama Bhasin,3 Susan Zolla Pazner,4
Suman Laal,4 and Kalpana Luthra1
1
Department of Biochemistry; 2Department of Medicine; 3Blood Bank, CN Centre, All India Institute of
Medical Sciences, New Delhi, India; and 4Veterans Affairs Medical Center, NYU School of Medicine New
York, NY
Background: Clade C viruses are primarily responsible for the HIV pandemic. Antibodies which
bind HIV and effectively neutralize the virus are needed to reduce the viral load. These antibodies are
targeted to different regions of the viral envelope such as the membrane proximal external region of
gp41, the CD4 binding on gp120, complex glycans on gp 120, the CD4 induced epitopes in and around
the gp120 bridging sheet and the V3 loop of gp120. The present day novel immunization approach is
directed at designing an ‘‘immunofocussing vaccine’’. In order to enhance the quality and/or quantity
of neutralizing antibodies, the V3 loop of the gp120 has been focused. Anti V3 antibodies have been
shown to efficiently neutralize primary isolates of HIV.
Methods: In order to characterize the immune response elicited by HIV-1 infected patients visiting
AIIMS, New Delhi, we detected the presence of anti V3 antibodies in their plasma and determined the
neutralizing efficiency. Neutralizing activity of the patient plasma at different dilutions was measured
using the TZM assay against a primary isolate raised. Presence of anti V3 antibody in the plasma of
these patients was detected using consensus C V3 peptide ELISA.
Results: Out of the 29 plasma samples, 17 showed .90% neutralization, 2 showed neutralization
between 70–80%, 6 showed ,50% neutralization, while 4 plasma samples revealed increased
infection in comparison to the virus control. All the plasma samples were found positive for V3
antibodies in comparative amounts based on the A405 obtained.
Conclusions: Our study suggests the presence of immunogenic epitopes which are more exposed
and therefore responsible for eliciting neutralizing antibodies, in majority of the patients. Besides V3,
there may be other immunogenic determinants responsible for efficient neutralization of HIV and this
need to be characterized. These sera are being tested further to check for neutralization of different
primary isolates raised in the laboratory to determine if they exhibit a broad cross-clade neutralization
of different viruses.
235 Quality ART Scale Up Through Regionalization of
Laboratory Services In Nigeria
C. Ezeaku, J. Farley, P. Dakum, T. Croxton, N. Constantine, A. Abimiku, and

W. Blattner
Issues: Setting up of standard lab infrastructure and provision of free laboratory tests by PEPFAR
has improved health care delivery system and services to PLWHAs in Nigeria. We here describe the
challenges and successes of the tier laboratory infrastructure established by the Institute of Human
Virology - University of Maryland PEPFAR funded AIDS Care and Treatment in Nigeria (ACTION)
program to support the diagnosis, care and treatment of HIV infected Nigerians.
Description: The ACTION program currently treats over 80,000 and cares for another 150,000 HIV-
infected Nigerians; also, about 300,000 are screened. To support this, the laboratory infrastructure
expanded from 12 to a regionalized tier system with 22 tertiary, 25 secondary and 12 primary
laboratories all linked with appropriate oversight and referral system. Choice of laboratory equipment
was based on ease of use, in country vendor/service provider, ease of purchase of reagents, and overall
manufacturer’s support.
Implementation included:
1) Decentralization of laboratory field services into regions with central oversight
2) Detailed site assessments/renovations
3) Step wise site activation in HIV Rapid Testing, Pregnancy Testing, Manual CD4 Count, HbAg
screening, CyFlow SL (tertiary) or Counter (secondary) for CD4, Sysmex KX 21N for Hematology, Vitros
V250 (tertiary) DT 60 II (tert/sec) or Reflotron (sec/prim) for Chemistry assays.
4) Monitoring and mentoring of site personnel through an aggressive QA/QC program.
5) Hub and spoke linkage between comprehensive and satellite sites.
Lessons learned:
Tier lab network with adequate mentoring and referral led to a successful lab expansion to 59 (22
tertiary, 25 secondary and 12 primary) sites performing HIV Serology, CD4, Pregnancy, Hematology,
Chemistry, Hepatitis B, Cryptococcus, and TB tests. Also, 57 HCT/PMTCT sites performing HIV Serology
Tests.
Improved proficiency at sites from 40–60% to 80–95%.
Recommendations:
Regionalization and tier lab system is critical for scale up.
Defined roles for lab personnel creates a good network to support rapid scale up.
QA/QC program involving site monitoring is critical for sustained high quality lab results.
236 T-Cell Receptor (TCR) Activation Mediates Efficient
and Sustained HIV Transcriptional Elongation and
Initiation through Multiple Signal Pathways
Joseph F. Hokello
Department of Molecular Biology and Microbiology, CWRU School of Medicine
The persistence of HIV in a latent pool during intensive drug therapy remains the single greatest
obstacle to successful HIV eradication. T-cell receptor (TCR) activation induces multiple signal
transduction pathways resulting in activation of several factors previously reported to regulate HIV
transcription such as NF-kB, NF-AT and AP-1. We analyzed the contribution of each of these
transcription factors in the reactivation of latent HIV using Jurkat T-cell clones harboring single latent
HIV proviruses. Jurkat T-cells were activated through the TCR by co-treatment with anti-CD3 and CD28
antibodies. The kinetics of proviral reactivation following mobilization of NF-kB by treatment with TNF-
a which induces NF-kB as the major transcription factor is characterized by an initial 4 hr lag phase
where elongation is restricted until new Tat is synthesized. In contrast, TCR activation induces efficient
and sustained HIV transcriptional elongation of latent proviruses. At initial time points, RNAP II is
recruited to the HIV LTR in parallel to NF-kB induction. However, during latter stages of activation
when AP-1 is induced, both NF-kB and AP-1 co-assemble on the promoter. Treatment of cells with
MAPK inhibitor PD98059 blocked AP-1 mobilization and recruitment to the promoter and these result
in a remarkable loss of RNAP II elongation as demonstrated by strong reduction in RNAP II levels
present downstream of the promoter. Studies using cyclosporine and LTR mutants demonstrate that
NF-AT does not play a major role in activating HIV transcriptional elongation or initiation when NF-kB is
present. However, in the absence of NF-kB, a combination of NF-AT and AP-1 can stimulate
transcriptional initiation. In addition to regulation of elongation by AP-1, we have found that P-TEFb is
‘‘pre-activated’’ by a novel pathway following TCR activation. During the first hour after TCR activation,
there is enhanced P-TEFb recruitment and RNAP II processivity which can be blocked by ERK inhibitor
U0126. We conclude that HIV transcription is a highly dynamic process which is tightly regulated by
multiple and complimentary signal pathways during TCR activation.
237 N-linked glycans on HIV-1 gp120 are critical
determinants for the recognition of CD4 helper T cell
epitopes
Hualin Li
VA Medical Center, New York, NY
Background: The heavy glycosylation of HIV-1 envelope gp120 shields this important antigen
from recognition by neutralizing antibodies and cytolytic CD8 T cells. However, very little work has
been done to understand the influence of glycosylation on the generation of MHC class II-restricted
gp120 helper epitopes and their recognition by the CD4 T cells.
Methods: Three glycans in the C4 region of gp120 (linked to N406, N448 and N463) and three
glycans in the C2 region (linked to N197, N230 and N234) were removed individually or in combination
by N-to-Q substitutions. These glycans were selected because they flank Ôhot-spotsÕ where many of the
helper gp120 epitopes are located. The wild type and mutant gp120 proteins were produced in CHO
cells and tested for recognition by CD4 T cell lines specific for different gp120 epitopes. The effect of
the N-linked glycan removal on proteolytic processing of gp120 was evaluated by trypsin digestion
and mass spectrometry (MS) analysis.
Results: The mutant proteins lacking the N448- or N230-glycans did not effectively stimulate CD4
T cells specific for the nearby epitopes, although the same mutants were recognized well by CD4 T cells
specific for more distant epitopes. Notably, both of these glycans were not part of the helper epitopes
and were located 2 amino acids downstream from the core epitopes. Data from trypsin digestion and
MS analyses demonstrated that the N448-glycan removal impeded the proteolytic cleavage of the
nearby C4 region, without affecting more distant sites. In contrast, the N230-glycan had no discernible
effects on the trypsin digestion of the neighboring C2 region, but the exact enzymatic processing
necessary for the generation of the C2 epitopes remains unclear.
Conclusions: The presence or absence of a single N-glycan on HIV-1 gp120 can dramatically alter
CD4 T cell recognition of this antigen. While the mechanisms by which the N-glycans affect CD4 T cell
recognition of gp120 are not yet fully understood, the glycans may dictate the local conformations of
the nearby gp120 regions and therefore influence the way CD4 helper epitopes are processed and
generated. Hence, N-linked glycans are critical determinants that can profoundly influence CD4 T cell
recognition of HIV-1 gp120.
238 Comparative Evaluation of the Performance of
Abbott m2000rt Real-Time HIV-1 Assay for
Measurement of HIV-1 Plasma Viral Load
M. Vidya,1 S. Saravanan,1 S. Kartik,1 K. Venkatesh,2 P. Balakrishnan,1

N. Kumarasamy,1 K. G. Murugavel,1 Sunil S. Solomon,1 Suniti Solomon,1
and Kenneth H. Mayer2
1
YRG Centre for AIDS Research and Education, Voluntary Health Services, Taramani, Chennai, India;
2
Brown University, RI, USA
Background: Measurement of HIV-1 plasma viral load has become a corner stone in the clinical
management particularly in the resource-limited settings. The objective of the present study is to
evaluate the performance of recently introduced ABBOTT Real Time HIV-1 PCR assay (ABBOTT
m2000rt) in comparison with the standard, COBAS Roche HIV-1 Monitor version 1.5 assay.
Materials and Method: A total of 50 stored (270°C) HIV-1 positive plasma samples with known viral
copies/mL {,400 not detected (n = 10); ,400 detected (n = 10); .400 – 5000 (n = 10); .5000 – 50000
(n = 10) and .50000 – 500000 (n = 10)} tested using the Roche HIV-1 Monitor ver. 1.5 assay and 20 HIV-1
low-risk seronegative (SN) samples were randomly used for the validation. Plasma RNA was extracted and
concentrated using ABBOTT manual specimen preparation, which uses magnetic particle technology to
capture nucleic acid extracted from 600 uL of plasma (detection range of 40– 100000 copies/mL). Plasma
HIV-1 RNA levels were log10 transformed before being subjected to statistical analysis. Averages were
statistically compared using the paired Z test. Linear regression and Bland-Altman were done to assess
the agreement between these two assays.
Results: The good correlation and agreement between ABBOTT Real Time and Roche was
observed. Although the difference in viral load determinations was positively skewed in favor of the
Roche assay, the difference in each PVL category, between the assays was within 0.5 log and the results
were well correlated [R = +0.89 (P , 0.05)]. Bland-Altman analysis found 95% of the samples within
clinically acceptable limits of agreement (21.001 to 0.52 log copies/mL). The sensitivity and specificity
of ABBOTT Real Time PCR detecting the viral copies of more than 1000/mL is 100% and .87%,
respectively. The specificity of 100% by ABBOTT Real Time PCR with the testing of SN for HIV-1/2 plasma
samples was observed.
Conclusion: The new ABBOTT RealTime PCR plasma viral load assay is sensitive and meets the
requirements for a wide dynamic range and providing higher throughput reliable quantification of HIV-1
RNA in plasma specimens.
239 Barriers to timely initiation of antiretroviral therapy
among HIV infected children admitted to Mulago
Hospital Paediatric wards
Dr. Eleanor Namusoke

Joint Clinical Research Centre, Kampala, Uganda
Background: The median survival of HIV infected children in the absence of treatment is 2 years;
thus early initiation of ART is desirable.
In Uganda, ART is now being provided free of charge. However, some children still do not access
the treatment. No study has been done in our setting to establish what could be the possible barriers
to timely initiation of ART.
Objective: To determine the proportion of children eligible to start antiretroviral therapy who have not
yet started by 4 weeks after discharge, and the factors that hinder access to timely initiation of antiretroviral
therapy in these children.
Methods: A cross sectional study was conducted on the paediatric wards at Mulago Hospital and
the Paediatric Infectious Disease Clinic (PIDC). 113 newly diagnosed HIV infected children in WHO
clinical stage 3 and 4, aged 3 months to 12 years were recruited. Questionnaires to assess factors
affecting timely initiation of ART were administered at recruitment and at 4 weeks after discharge.
Home visits were also done. Qualitative data was collected using focus group discussions, and in-
depth interviews.
Results: The proportion of children who were eligible to start antiretroviral therapy but had not
yet started by 4 weeks after discharge was 97.3% (110/113). Incomplete adherence counselling, default
and death were the major barriers that prevented initiation of ART.
Conclusion: 97.3% of the children had not started ART by 4 weeks. Caregiver factors were the
main barriers that prevented initiation of ART in these children.
240 Patient Retention in a University Hospital-based ART
program in Uganda
Ouma Joseph
Faculty of Medicine, Makerere University, Kampala, Uganda
Background: Access to antiretroviral therapy (ART) in resource limited settings has increased over
the past five years. ART programs report good adherence but these data are usually generated from
patients who are retained in care. Documentation on long term follow up of patients on ART is limited.
We evaluated the two-year retention of patients receiving ART from 12 clinics (8 in urban and 4 in rural
settings) supported by Mulago-Mbarara Teaching Hospitals’ Joint AIDS program (MJAP). MJAP
provides ART to over 14,000 patients. Patients are initiated on ART if CD4+ counts ,200 cells/mm3 or
WHO stage III or IV. The preferred first line ART regimen is Nevirapine and Combivir containing.
Patients are given monthly appointments for ART refills, clinical evaluation and counselling. Treatment
supporters are encouraged. Telephone calls and/or home visits are used to track patients who
miss appointments.
Methods: We retrospectively analyzed data for 6,067 ART na¨ve adult (18+ years) patients initiated
on ART between November 2005 and November 2007. We assessed predictors for retention on ART
using Cox proportional hazard model.
Results: The median CD4+ count at ART initiation was 132 cells/mm3 (IQR: 56,193). Total attrition
was 15% for urban based sites and 7% for rural based sites. Death rate was 5.4% overall (5.7% urban
and 4.9% rural). Overall transfer out was 5.5% (6.5% urban and 2.3% rural), and lost to follow up 2.2%
(2.8% urban and 0.2% rural). Estimated attrition rate was 30.5 per 100 person years over 2.4 years of
follow up. Nevirapine based ART regimen, Female sex, weight .50 kilograms, CD4+
counts .200cells/mm3, and rural clinics were associated with lower loss to follow-up, P-value ,0.001.
Conclusion: Retention was high and comparable to other ART programs in Africa. Loss to follow-
up is higher in urban sites, possibly related to mobility especially of male patients. Enhanced patient
tracking and support may be necessary for patients accessing ART services in the urban areas and
those with baseline CD4 count below 200 cells/mm.
241 High Incidence Cohort of IDUs Infected with HIV with
Low Genetics Diversity for HIV Vaccine Efficacy Trials
Sergey Verevochkin
St. Petersburg, Russia
Background: In the previous study (NIH HPTN 033) HIV incidence was determined in the cohort of
injection drug users (IDU) from St. Petersburg, Russia. Between March and December 2002, 898 IDU
were screened for HIV and 270 (30%) were HIV positive. 520 uninfected IDU were enrolled in the cohort
study. The retention rate at the 12-month follow-up period was 80%. HIV-1 incidence rate was 4.5 per
100/PY. Similar incidence (4.8%) was obtained using STARHS detuned method.
Methods: There are two current studies at the Biomedical Center among high-risk groups for HIV
infection. The cross-sectional study (SATH-CAP) used the respondent-driven methodology to recruit
DUs, their sex partners, and sex partners of sex partners through chain referral. Sera from HIV positive
study participants were tested with Calypte HIV-1 Incidence EIA. Incidence was calculated in
accordance with CDC consensus formula (comes with manual). Interview data were used for
confirmation of infection date. The Randomized Controlled Trial of Russia IDU Peer Network HIV
Prevention Intervention includes follow-up at 6 and 12 month. Incidence data based on
seroconvertions during follow-up visits were compared with Calypte assay and interview data.
Results: During 2005–2006 years 419 from 909 IDU patients were found positive for HIV-1
antibodies in EIA and confirmed by WB (46,1%). 72 of them were negative on Calypte assay. Interview
showed that 15 patients were found positive for HIV-1 antibodies at least 6 months before Calypte test.
In addition, follow-up sera from 6 patients were negative on Calypte assay at least 6 month after first
Calypte test. So, only 51 specimens were classified as recent infections, which gives 21, 2 per 100 p-y.
The incidence data from the cohort study performed during the same period showed 19.6 per 100 p-y.
Conclusion: These two studies show that incidence of HIV infection among IDU has substantially
increased during the last several years. Different approaches (the cohort study and laboratory testing
for incidence cases) produced close results of 19.6 and 21.2 percent incidence. This is the highest
published incidence of HIV infection to our knowledge. The current situation of high HIV incidence
rates suggests the need for implementation of intense prevention strategies. In the context of
a homogeneous infecting HIV strain the situation is also ideal for mounting efficacy trials of HIV
vaccines among IDUs in St. Petersburg.
242 Mycobacterium avium KatG protein (MAV_2753):
a putative candidate for the serodiagnosis of MAC
disease
Kapil Gupta,1 Ajay Wanchu,2 Romica Latawa,1 S. Laal,3 G. K. Khuller,1 and

Indu Verma1
1
Department of Biochemistry; 2Department of Internal Medicine, Postgraduate Institute of Medical
Education and Research, Chandigarh, INDIA; 3Department of Pathology, NYU Langone Medical Center,
New York, USA
Disseminated Mycobacterium avium complex (MAC) infection is a severe complication of

advanced HIV disease. Recent studies suggest that Mycobacterium avium, a member of M. avium
complex (MAC) is responsible for about 5–7% of the mycobacterial infections in HIV+ patients in India.
Currently available diagnostic methods for differentiation of M. tuberculosis (M. tb) and MAC infections
require culture of bacteria, which is not routinely available in India and is time-consuming and
complex. A simple and rapid diagnostic assay that can identify M. avium disease and can discriminate
between M. tb and M avium is urgently required. Present study was aimed to identify a M. avium
secretory protein as a candidate serodiagnostic marker for MAC bacteremia in HIV patients. Based on
a combination of biochemical and immunological approaches, we have identified the ;81 kDa M.
avium KatG protein to be immunodominant antigen that is recognized by antibodies in 90% of HIV-
infected patients who were confirmed to have disseminated MAC infection on the basis of blood
culture. In contrast, no antibodies to this protein were detected in sera from TB patients, healthy
subjects, or HIV-infected subjects. These data suggest that the M. avium KatG protein can be useful in
designing a simple and specific serodiagnostic assay to rapidly diagnose the bacteremia due to M.
avium in AIDS patients. Such a diagnostic test could be of great help to clinicians in determining the
therapeutic strategies for HIV-infected patients suffering from mycobacterial diseases.
243 The Use of HIVQUAL as a Quality Improvement Tool
for a Large Scale HIV/AIDS Public Health Program –
U. Yakubu, U. Gebi, M. Babamaiyaki, I. Okoye, K. Falayajo, P. Dakum, J. Farley,

W. Blattner, W. Etiebet, A. Zoakah, and M. Charurat
Institute of Human Virology, Abuja, Nigeria
Background: With 67, 543 patients currently in care, the ACTION project is a comprehensive
HIV/AIDS care and treatment program sponsored by The Institute of Human Virology, University of
Maryland School of Medicine since March, 2005. Initial efforts at rapid scale-up in order to address
unmet demands for HIV/AIDS services within the country have been successful and the organization
has decidedly shifted its focus from acquiring more patients to improving the quality of clinical care
and other services that are being delivered to our patients who are already in care.
Design/Methods: The ACTION project provides comprehensive HIV/AIDS care at 40 sites. Random
sampling was used to select 23 sites spread across the six geopolitical zones of the country for
a Quality Assessment Exercise spanning 2 weeks. The methodology employed the use of HIVQUAL
indicators to assess the level of clinical care. A total of 5 tools were administered to assess the overall
quality of care. These included a random chart review, administered questionnaires, focus group
discussions, and site observation charts. For the random chart review a case list of active patients (i.e.
any patient who has had at least one clinical visit in the last 6 months) was generated by the clinic.
Sample size for each clinic was determined by active caseload. A randomized sample of patients was
generated from the case list using the HIVQUAL sampling table.
Core indicator abstraction which included Continuity of care, CD4 count monitoring, TB screening
and assessment, Cotrimoxazole preventive therapy, Weight monitoring, Prevention education,
appropriate ARV therapy and Adherence assessment were extracted from medical records and charts.
Data was entered into the software and reports were generated automatically. A special element of
this exercise was the specific auditing of Ôkidney function monitoringÕ by scheduled creatinine
assessments of patients who had been newly initiated on TDF–based regimens with at least 12 months
of follow-up after ART initiation. Questionnaire administration using the second tool involved
ÔInterviewsÕ with the ARV focal person, the Matron of the clinic, one junior medical officer and one
nursing officer in order to obtain their subjective impressions of the quality of key clinical activities that
243 The Use of HIVQUAL as a Quality Improvement Tool
for a Large Scale HIV/AIDS Public Health Program –
(continued)
were occurring in their clinics. The third tool involved the conduction of Focused Group discussions
with separate focus groups including Physicians, Nurses, M & E officers, Record staff and the cleaning
and messenger staff. The fourth instrument was the employment of a ÔQuality of care Observation toolÕ
which assessed the more general aspects of clinic flow and clinic conditions including patient waiting
time, sitting space, physician to patient ratio on clinic day and activity coordination. The ready
availability of relevant guidelines, SOP’s, cleanliness of the clinic environs, use of a team system and the
presence of functional basic clinic equipment was also incorporated into this observation tool.
Results: The results showed that compliance with protocol for laboratory testing was highest at
baseline with CD4 counts, Full blood counts and liver function tests being ordered for virtually all of
the patients. TB symptom screening was also high at baseline (85%). Ordering of follow up
investigations however depreciated progressively with time, falling to about 60% at 6 months when all
the sites are averaged together, and to about 50% at 12 months. Creatinine assay request was over
98% for most sites at baseline but dropped sharply to less than 40% across the board at 6 months and
to less than 10% at 12 months. Patients being placed on Cotrimoxazole prophylaxis when their CD4
count was below 350 was about 80% while the correct and complete filling of clinical evaluation forms
had a compliance of 45% across all sites. Weight measurement, which is carried out by nurses in the
clinic, was the quality indicator with the highest level of compliance, consistently showing .98%
documentation for all sites, across the entire study period. This particular finding of high compliance
suggests that this capacity could be exploited in graduated task shifting processes which will relieve
other overburdened staff, such as doctors prescribing ARV’s.
Challenges and Lessons Learned: Feedback meetings were carried out with all the site project
staff to share study findings and solicit their opinions. The most important and consistent obstacle to
provision of quality service was Manpower. Critical lab investigation benches such as CD4 count and
chemistry were manned by only one staff. If such a staff was indisposed or resigned there was
immediate interruption of service which was a common occurrence. Doctor to patient ratio on clinic
day was also suboptimal in most sites with one doctor commonly attending to more than fifty
patients. Task shifting to the nurses who achieved a high compliance on weight monitoring should be
considered as a result of the findings of this exercise. In conclusion it can be said that the HIVQUAL
approach to quality assessment can serve as a very useful tool for quality monitoring and quality
improvement for large-scale public health programs such as IHVN’s ACTION PROJECT.
244 Mannose Binding Lectin and its variants: susceptibility
to HIV-1 and Schistosoma haematobium infection in
a rural Zimbabwean community
Rutendo B. L. Zinyama-Gutsire
National Institute of Health Research Ministry of Health and Child Welfare Zimbabwe
Introduction: The last decade has seen an emerging interest in Mannose Binding Lectin (MBL) due
to its role in innate immunity. Low serum concentrations of MBL are associated with increased
susceptibility to infections. We investigated the role of MBL in HIV-1 and Schistosoma haematobium
coinfected and uninfected individuals.
Objective: To determine levels of MBL in plasma of HIV-1 and Schistosoma haematobium coinfected
and uninfected individuals in a rural Zimbabwean community and to test whether MBL concentrations or its
genetic variants are associated with increased risk of HIV-1 infection and death.
Methods: Through a cross-sectional survey, during which HIV-1 and schistosomiasis screening
were done, a cohort of 379 participants was established. Outcome measures consisted of HIV-1 and
schistosomiasis status and levels of MBL in plasma at baseline. Non-parametric statistical methods
were used to assess the relationship of MBL levels or its genetic variants, with HIV-1 and S.
haematobium infections.
Results: A total of 379 adults formed the established follow up cohort comprising of 74 (19, 5%)
men and 305 (80, 5%) women. Mean age was 33.25, range 17 to 62 years. There was no difference in the
mean MBL plasma concentration of HIV-1 positive and HIV-1 negative participants (P = 0.066) but there
was a significant difference in the mean MBL plasma concentration of S. haematobium positive and
negative participants (P = 0.0173). There was a significant difference in the mean MBL plasma
concentration between HIV-1 and S. haematobium coinfected and uninfected individuals (P = 0.087). The
distribution of the three genotypes (A/A, A/O and O/O did not differ between the HIV-1 infected and the
healthy controls and neither between those S. haematobium infected and uninfected. When HIV-1 S.
haematobium infected participants were analysed separately, there was a significant difference in MBL
levels between the three genotypes. In HIV-1 positive participants, there was no difference in rate of
death between those with normal MBL compared to those with variants, log rank test 0.274.
Conclusions: The results of our study show that HIV-1 infection had no influence on MBL plasma
concentrations but that there were significantly higher levels of MBL in S. haematobium infected than
those uninfected. The distribution of the three MBL genotypes (A/A, A/O and O/O did not differ
between the infected and the healthy controls. We conclude that MBL homozygosity nor
heterozygosity is not associated with increased risk of HIV-1 infection in this Zimbabwean cohort
but there was increased production of MBL in S. haematobium infection. Among those HIV-1 positive,
those with variant MBL were not at increased rate of death.
245 Outcome of early infant diagnosis in exposed infants
in Northern Nigeria
O. D. Adegoke, Z. Basir, J. Jumare, S. Sani, R. Enzama, S. Peters, A. Abimiku,

and W. A. Blattner
Background: The rate of vertical transmission of HIV-1 in the Northern Nigeria was 35%. This
prompted the intervention of PMTCT and Early Infant Diagnosis program in Northern Nigeria. Aim
Evaluating the outcome of early infant diagnosis of HIV in exposed infants in Northern Nigeria and the
rate of mother to child transmission of HIV in the region.
Method: A total of 1236 HIV exposed infants from the Northern Nigeria, (Kano, Kaduna, Katsina,
Jigawa, and Sokoto states of Nigeria) who had Dry Blood Spot (DBS) HIV-1 DNA Polymerase Chain
Reaction test were enrolled for the study. The age of enrollment was between 6 weeks and 18 months
with mean age of 7 months. Roche HIV 1 DNA version 1.5 method was used for analysis.
Result: Total number of women who had PMTCT was 1,036 (83.8%), 182 (14.7%) had no PMTCT
and 18 (1.5%) women had unknown PMTCT status. 124 (10%) children were HV-1 positive and 1112
(90%) were HIV-1 negative. 109 (87.9%) of those whose mothers had PMTCT were HIV-1 positive and
those whose mother did not have PMTCT had 15 (12.1)% HIV-1 positive results. 520 (42%) children
were not breast fed NBF and 495 (95.6%) of this children were HIV-1 DNA PCR negative and 25 (4.4%)
children were HIV-1 DNA PCR positive. 437 (84%) of the NBF had PMTCT(PMTCT +) while 70 (16%) had
no PMTCT (PMTCT2). 345 ceased breast feeding (BF Ceased) before 6weeks and 18 months and after
their babies were tested for HIV. 285 (82.8%) children from this BF ceased mothers were tested HIV-1
DNA PCR negative and 60 (17.2%) was HIV-1 DNA PCR positive.
Conclusion: Through the PMTCT and EID interventions, HIV-1 prevalence rate among the children
of exposed mothers was dropped from 35% to 10% in Northern Nigeria. Attitude of women to
absolute breast feeding in the Northern Nigeria have been changed for the benefit of the children.
Through the EID program, children’s status are known and early ARV intervention is instituted. These
result in decline mortality rate and more and more children are living meaningful lives in this part of
the world.
246 Interface of Public Health Implementation and
Research – Challenges and Prospects – IHVN/ Action
Project Experience
O. Akinwande, Patrick Dakum, J. Farley, U. Gebi, C. Adebamowo, A. Abimiku,

M. Charurat, and W. Blattner
Background: The first case of HIV was described in Nigeria in 1986 with a characterization of the
predominant subtype as AG recombinant and since then the prevalence has risen from an ANC
Sentinel survey of 1.8 in 1991 peaking to 5.8 in 2001 and declining in 2005 to 4.4. With a population of
140m Nigeria has the second highest burden of HIV globally after South Africa. There are an estimated
5 million Nigerians currently living with the virus. In 2002 the Nigeria Government with assistance from
IHV and other Partners commenced a pilot ART program that had a total of 10,000 people on therapy
by 2004.
Public Health Implementation: With PEPFAR Funding in 2004, the IHV commenced implementing
the ACTION Project which currently supports 52 Health Facilities ranging from primary to tertiary
(Teaching Hospital) sites. As at Jun 2008, ACTION Project has 57399 person receiving ART out of which
3711 are children below 15 years and 1310 are pregnant women. Laboratory and Data Management
capacities have been built.
Research: IHVN Research department currently coordinate research efforts focusing on assisting
collaborators in site coordination of research staff, laboratory assays including sample transfers, data
management and coordination of an Investigators working group. With Fogarty Funding, IHVN
supported the training of 38 persons from 8 hospitals within its network on Ethical Issues related to
research. IHVN has also facilitated the establishment of credible CITI certified IRBs in most of its
collaborating sites.
Challenges including lack of trained research assistants, committed and dedicated research staff,
CITI certification for a number of site IRBs and power are issues that are being addressed.
Conclusions: With about 5 internationally certified researches ongoing, Several accepted
abstracts in international conferences and meetings and a number of proposals in final stages, ACTION
presents a model for the interface of international public Health implementation and research and an
opportunity for collaborators interested in international research both basic science and public health
to provide evidence based answers to emerging public health questions in PEPFAR and Global
Fund Implementation.
247 The effect of chronic alcohol exposure in HIV/AIDS
patients on antiretroviral drugs (Triomune30 – AZT/
3TC/NVP) in Uganda
Dr. Godfrey Sande Bbosa, J. Ogwal-Okeng, W. W. Anokbonggo, and

B. D. Kyegombe
Department of Pharmacology and Therapeutics, Faculty of Medicine, Makerere University,
Kampala, Uganda
Alcohol is a major business worldwide and Uganda is ranked among the top alcohol consumers
globally. Alcohol consumption is known to be a serious risk factor in the HIV transmission due to its
effect on the CNS where it interferes with the judgment and decision making processes in the brain
thus making people to engage in unsafe sex. Alcohol may also interfere with the drug metabolizing
enzymes in the body like the ARVs hence leading to therapeutic failures if enzymes are induced or
toxicity if the enzymes are inhibited. It may also affect the adherence of the HIV/AIDS patients on ARVS
by forgetting to take their drugs on time. However, many of these patients are observed to consume
alcohol despite being told to stop the habit but with no scientific data on alcohol, HIV and ARVs
interaction. The longitudinal cohort study (still on-going PhD work) is being conducted to determine
the effect of the chronic alcohol exposure in HIV/AIDS patients on triomune-30 in Uganda. This study is
in the final stages of completion and is aimed at determining the triomune-30 drug plasma
concentrations (Pharmacokinetics), the viral load, CD4 count, CBC and the LFT parameters in these
patients exposed to chronic alcohol (therapeutic response).The outcome of this study will enlighten
researchers on the alcohol, HIV and ARVs interaction which will be important in the policy formulation
and guiding the clinicians on how to use these drugs instead of just switching drugs because of the
therapeutic failure or toxicity like lactic acidosis associated with triomune-30 as observed in various
ART clinics in the country that might lead to increased resistance to these drugs.
248 Telemedicine in Peru: training physicians
responsible for the administration of highly active
antiretroviral therapy (HAART) in a developing
country
Katiuska Castillo,1 Raú Gutiérrez,1,3 Leslie Soto,1 Alfonso Silva-Santisteban,1

David Iglesias,1 Alberto Guerra-Garcı́a,1 Carlos Kiyan,2 Carlos Seas,1
Juan Echevarrı́a,1 Ciro Maguiña,1 and Eduardo Gotuzzo1
1
Instituto de Medicina Tropical Alexander von Humboldt, Universidad Peruana Cayetano Heredia, Lima,
Peru; 2Facultad de Medicina, Universidad Peruana Cayetano Heredia, Lima, Peru; 3Fogarty-NIH
International Training and Research Program in HIV/AIDS and TB University of Miami, Miller School of
Medicine
Issues: With support of the Global Fund, free HAART was implemented in Peruvian public
hospitals and health centres in 2004. This effort included the conformation and training of
multidisciplinary teams (Physicians, nurses, psychologists, social workers) responsible for the care of
people living with HIV/AIDS throughout the country. Physicians working in remote regions would
administer HAART for the first time and had to be thoroughly trained with limited resources.
Description: The Telemedicine Unit of the Alexander von Humboldt Institute of Tropical Medicine
developed a long-distance course on ‘‘Use of antiretrovirals’’ using an electronic platform as
‘‘classroom’’ Participants could access internet in their local hospitals or in public cabins which are
widely distributed throughout Peru. The course took place from April–December 2005, covering 12
modules each lasting 2 weeks. Topics included adverse effects, antiretroviral schemes, resistance,
vertical transmission, ethics, among others. (see www.upch.edu.pe/tropicales/telemedicinatarga). Each
module consisted of a pre and post test, presentation, discussion forums, clinical cases and summaries
of scientific articles.
Lessons Learned: Sixty-eight physicians from 21 of the 24 Peruvian Regions started and 81% (n =
55) successfully completed the course. While 73% (n = 41) were already administering HAART, the
others were waiting to be programmed by the Ministry of Health. Forty-nine physicians participated in
an evaluation at the end of the course. They all pointed out the usefulness of the course in their daily
practice; 98% (n = 48) classified the course as good/very good and 88% (n = 43) stated that the
operational system was easy/very easy to use.
Recommendations: After the successful implementation of a long-distance course about use of
antiretrovirals, this course will be repeated for a broader target group of Peruvian physicians
administrating HAART. Telemedicine has shown to be a powerful and practical tool for continuous
training of health care personnel throughout the country and for the reinforcement of quality care to
people living with HIV/AIDS.
249 Efficacy of a Behavioral Intervention to Reduce HIV
Risk among Female Sex Workers in Urumchi, China
Lin Han
Division of Policy and Information, NCAIDS, Beijing, China
Objectives: To assess the effectiveness of the behavioural intervention among FSWs in minority
area of China.
Methods: An experimental study using convenience sampling was conducted in Urumchi, capital
of Xinjiang Uygur Autonomous Region. Participants attended a series of three biweekly 60-minute
sessions and a three-month follow-up. The intervention program was culturally specific, addressing
HIV-related knowledge, safer sexual practices, which consisting of education session, discussion
session, counseling session and condom distribution.
Results: A total of 229 brothel-based FSWs were recruited and 164 of the original participants
were interviewed at the three months follow-up. As for the HIV transmission mode, the variables of
kissing, sharing food, toilet, shaking hands and mosquitoes were statistically significant before and
after the intervention (P , 0.001). Exact test found FSWs had more positive attitudes on health care,
work ability of PLWHA and their own responsibility on AIDs prevention (P , 0.05) in follow-up survey.
Consistent condom use and condom use during last sexual encounter with primary and other partners
last month were significantly different before and after the intervention (P , 0.05).
Conclusions: The intervention program consisting of culturally specific education, discussion and
counselling sessions can be effective in changing self-reported, high-risk sexual behaviours with
primary and other partners.
250 Establishment of new Jurkat cell line Stably
expressing HIV-1-Tat
Mohamed Ali Jarboui, William W. Hall, and Virginie W. Gautier

Centre for Research in Infectious Diseases, University College of Dublin, Dublin, Ireland
The trans-activator Tat protein is a critical regulatory factor for HIV-1 replication. We have
established a new Jurkat-Tat cell line, which constitutively expresses HIV-1 Tat.
Using retroviral gene delivery, we infected Jurkat cells with a VSV-G pseudo-typed virus. The virus
was produced using Pheonix-GP cell line co-transfected with a VSV-G plasmid and the pCeMM-NTAP-
Tat(GS)-Gw plasmid. HIV-1-Tat was cloned downstream a Tandem affinity purification (TAP) tag and
upstream an IRES region, followed by a GFP coding sequence. We confirmed the activity of the NTAP-
Tat protein using luciferase LTR reporter gene assay.
The GFP expression allowed us to monitor the integration and the expression efficiency of the
trans-gene and to isolate a heterogeneous population using FACS cell sorting. We enriched our
population up to 90 % GFP expressing cells, representing different integration events. The level of
expression of HIV-1-Tat was confirmed using Western-Blot analysis.
Interestingly, even after high passages number (35), our Jurkat-Tat cells continue to stably express
GFP and Tat. Furthermore, after preliminary observations, the expression of Tat does not alter the
viability of the cells when compared to Jurkat cells even at very high density cell culture.
The advantage of these Jurkat-Tat cells over several existing Jurkat clones is that Tat is expressed
without being fused to GFP, meanwhile the trans-gene integration and expression can be monitored
using the GFP expression level. Moreover, Tat is linked to a TAP tag which allows more efficient
purification of Tat. Ultimately, the Jurkat-Tat cells represent a good model to study the molecular
pathogenesis of HIV-1-Tat.
251 Migration, Pastoralists, HIV Infection and Access to
Care: The Nomadic Fulani of Northern Nigeria
J. Jumare,1 A. G. Habib,3 U. Gebi,1 A. Zoakah,1 P. Dakum,1 J. Farley,2 and

W. A. Blattner2
Institute of Human Virology Nigeria
Background: The burden of HIV infection among the nomadic Fulani of northern Nigeria is
unknown. Migration — a way of life for this population — is known to increase the rate of HIV
transmission and may limit individuals’ access to treatment and care. Many of Africa’s other traditional,
pastoral societies are similarly affected.
Objective: This paper explores cultural practices and factors among the Fulani that may influence
HIV transmission, vulnerability to infection, sustainability and challenges to treatment access, as well as
avenues and models for outreach services.
Method: An extensive literature search with cross-referencing was done, and relevant
publications on similar themes were reviewed. Three cases of Fulani nomads with HIV are presented
to illustrate the challenge of providing a care continuum as well as to demonstrate successes when
appropriate HIV interventions are employed.
Findings: Community mobility limits opportunities for counseling, testing and diagnosis, as well
as HIV-related care access and maintenance. Consanguinity and certain cultural practices among the
Fulani have clear amplification potential for HIV transmission. Treatment support through the use of
coaches and life partners improves adherence to antiretroviral therapy (ART). Existing programs for
nomads afford opportunities for absorption and integration of HIV services.
Recommendations: Nomadic communities should be provided with basic HIV-related services,
including risk-reduction education and methods, counseling and testing, ART, medication adherence
counseling, access to laboratory tests and health monitoring. These services should be taken to
nomadic communities using novel approaches such as mobile units, extension services, case
management, directly observed care, and treatment supporters linked to neighboring health facilities
in a hub-and-spoke model. Stronger collaborations are recommended between programs for nomads
and HIV services, and also between veterinary and public health services. Community participation
and leadership should be encouraged to ensure the sustainability of HIV-related care delivery.
Conclusion: More research is needed on the epidemiology and sociology of HIV infection and the
best ways to provide services to hard-to-reach nomadic populations. They may well constitute HIV
infection sanctuaries and should not be ignored.
252 Acceptance of and adherence to anti-retroviral
therapy in Tanzania: the influence of lipodystrophy
and of traditional medicine
Sajida J. Kimambo
Objectives: To assess the influence of lipodystrophy and of traditional medicine use on
acceptance of, and adherence to, anti-retroviral therapy (ART).
Design: Cross sectional study of HIV-infected subjects enrolled in a tuberculosis vaccine trial in
Tanzania.
Methods: ART eligible subjects who had either accepted (ARTpos) or refused ART (ARTneg) were
interviewed and examined to identify subjective and objective features of lipodystrophy and to assess
use of traditional medicine (TM).
Results: 52 ARTpos and 53 ARTneg subjects had median CD4 counts of 296/mm3 and 160/mm3
respectively. Subjective features of lipodystrophy were noted by 39 (75%) ARTpos and 11 (21%) of
ARTneg subjects (P = 0.0001). Median waist to hip ratios in males were .98 in ARTpos and .92 in ARTneg
subjects (P = 0.04); in females 0.90 vs 0.83 (P = 0.004). Concern about lipodystrophy was reported to
affect acceptance of ART by 19 (36%) of ARTneg subjects (P = 0.0001) and adherence to ART among 3
(6%) of ARTpos subjects. Traditional medicine was used by approximately 50% of subjects in both
groups and affected ART acceptance in 11 (21%) of ARTneg subjects and ART adherence in 7 (13%) of
ARTpos subjects.
Conclusion: Subjective and objective features of lipodystrophy are more common among ARTpos
than ARTneg subjects in Tanzania and appear to have a substantial effect on acceptance of ART, but not
on adherence to ART. Traditional medicine is common regardless of ART status, and appears to have
a modest effect on both acceptance of and adherence to ART.
253 Changes of HIV risk behaviors of heroin drug users
treated in methadone maintenance treatment
clinics in Guizhou province, China
Enwu Liu
Aims: To evaluate the HIV risk behaviors among heroin drug users who were treated in eight
methadone maintenance treatment clinics in Guizhou province, China.
Design and Methods: The study used a prospective cohort study design. Patients who had been
treated in the methadone clinic no more than two and half months were recruited into the cohort
beginning at June, 2006 and followed up until June, 2007. Face to face interviews were conducted to
collect baseline and follow up information on HIV risk behaviors. McNemar tests were used to test the
changes of HIV risk behaviors at baseline and follow-up.
Results: 1003 patients were interviewed at baseline and 469 patients were interviewed both at
baseline and follow-up for HIV risk behaviors. Comparing the 469 patients at baseline and follow up
the study found that needle sharing behavior in the last 30 days decreased from 1.3% to 0.2% (P ,
0.001); 5.5% patients had multiple sex partners in the last 30 days at baseline and 3.4% at follow up (P =
0.334); 6.4% had casual sex with casual sex partners or strangers in the last 30 days at baseline and
5.4% at follow up (P = 0. 030); when had casual sex in the last 30 days, condom use decreased from
39.1% at baseline to 19.6% at follow up (P = 0.039).
Discussion and Conclusions: Methadone maintenance treatment was effective for decreasing
needle sharing behaviors among heroin drug users. However, there was no significant decrease on HIV
risk sexual behaviors among heroin drug users treated in the methadone maintenance clinics. Condom
use with casual sex partners even decreased after patients treated in methadone clinics. Methadone
clinics should let patients know that methadone itself will not prevent HIV transmission.
254 Factors associated with development of
Opportunistic Infections among patients on ART in
Uganda
S. Muhumuza, J. Ouma, F. Semitala, E. Mbabazi, and M. Kamya

Mulago-Mbarara Teaching Hospital’s Joint AIDS Program (MJAP), Kampala, Uganda
Introduction: The introduction of Anti Retroviral Therapy (ART) has dramatically reduced the
incidence of Opportunistic Infections (OIs) among HIV positive people who have received ART;
however, potent ART does not replace the need for antimicrobial prophylaxis and treatment of OIs
among patients with severe immune suppression.
Background: Mulago-Mbarara Teaching Hospital’s Joint AIDS Program (MJAP) established under
the Faculty of Medicine, Makerere University in 2004 using a grant awarded by PEPFAR provides ART to
patients in 12 clinics basing on the eligibility criteria of CD4+ cell count ,200/mm3. By far, a total of
15,984 patients have been enrolled on ART.
Methodology: Retrospective cohort analysis on adult patients initiated on ART between
November 2005 and November 2007. The socio demographics, clinical and laboratory characteristics of
patients that developed opportunistic infections were studied.
Results: Data of 4,878 patients on ART was analyzed. 3,247 (67.2%) were females. Mean age 34.1
years (SD 8.5) and Mean weight 53.5 kg (SD 10.1). Prevalence of OIs 329 (6.7%); Oral candidiasis 106
(32%), Vaginal candidiasis 80 (24%), Kaposi’s sarcoma 40 (12%), Herpes simplex 28 (9%), Oesophageal
candidiasis 25 (8%), Tuberculosis 15 (5%), others 35 (11%).
WHO stage 3 &4, median CD4 count ,90/ mm3, not being married and having no formal
education were significantly associated with development of OIs among patients on ART (P , 0.001).
Sex, age, body weight, ART regimen and Karnofsky score were not associated with development of OIs.
Conclusion: The risk of OIs is higher among patients with a low CD4 count, high WHO clinical
stage, no formal education and in those that are not married.
255 HIV Related TB: Prospects and Challenges in a High
Burden Area, A Case Study of Isoniazid Prophylaxis
(IPT) Utilization
B. M. Musa, U. Gebi, K. Falayajo, P. Dakum, J. Farley, and W. Blattner

Background: The complex relationship between tuberculosis (TB) and HIV does not only have an
impact on the natural history of either diseases but also influence the design and implementation of
programs intended to deal with the needs of people affected by the diseases. Interventions to control
HIV-related TB comprise measures against both TB and HIV. However, in most settings, strategies to
accomplish this are only beginning to reach the sites, where their impact will be made and the
expectation of improving the outcome of both diseases realized. This study looked at the impact of
TB/HIV collaborative activities; its prospect and challenges.
Method: Data record of HIV+ patients accessing care were analyzed for information on TB prevention
and treatment; data were also extracted from the TB registers maintained and kept at the DOTS center for
tuberculosis treatment and HIV counseling and testing.
Result: Of the 60 Sites under UMD/ACTION project 23 (38.3%) have access to TB/HIV services. Of
the of 286,747 HIV+ patients; 18043 (6.3 %) were screened for TB; where 714 (4 %) had active TB. 1040
(6 %) of those patient in whom active TB has been ruled out were commenced on IPT. Of the 286747 that are
under care in the program 2443 (0.85 %) had HCT in DOTS clinics.
Conclusion: TB/HIV collaborative activity is feasible while trying to rapidly scale up HIV care and
treatment. There is also a modest achievement in treating TB/HIV co-infection and there is the goodwill
of clients to count upon while providing improved diagnostic capabilities and better access to care
and treatment. In spite of the effort made in expanding access to TB/HIV care a lot of work has to be
done to increase the uptake of IPT and the screening of HIV + patients for TB, as well as strengthening
the linkage between DOTS clinics and ART points of care.
256 Dyslipidemia in HIV-1 infected individuals following
generic efavirenz based HAART in a resource
constrained setting
Dr. Sundaram Muthu

YRG CARE, Voluntary Health Services, Taramani, Chennai, India
Background&Aims: Very little is known about the alterations of lipid profile among HIV-1 infected
individuals initiating antiretroviral therapy containing efavirenz based regimen in resource limited
settings. The study was aimed at evaluating the changes of lipid profile among HIV infected patients
with and without HAART in Southern India.
Methods: In a prospective study, 44 HIV-1 infected individuals (18 ART na¨ve and 26 on HAART)
were enrolled for this study at YRG Centre for AIDS Research and Education in Southern India. Lipids
and lipoproteins were estimated at baseline, 6 and 12 months using an Olympus AU 400 autoanalyzer.
Plasma HIV RNA concentrations were determined by the Cobas Amplicor (Roche molecular Systems,
NJ, USA) and Absolute CD4 T Lymphocytes count was determined using the Guava Personal Cell
Analyser (Guava Technologies, Inc., USA). Baseline characteristics are described using mean and
standard deviation and for non-normal data, median and interquartile range has been used. The
Wilcoxon Signed rank test was used to compare the change in lipid levels from baseline to 12 months.
A P value less than 0.05 were considered statistically significant. All analysis was done using SPSS v
13.0.
Results: Of 44 HIV-1 infected individuals, 65% were males in the treated group and were 61% in
the untreated group. At baseline, HIV infected individuals with and without HAART had a mean age of
32 and 30 years, BMI of 16.86 and 20.06 kg/m2 respectively. The median CD4 and Viral load in HIV
treated and untreated patients were 197 cells/mL (IQR: 125–279), 146000 copies/mL (IQR: 38300–
214000), and 492 cells/mL (IQR: 327–595), 9670 copies/mL. All patients were initiated on efavirenz
based HAART. From baseline to 12 months, patients on HAART had a significant increase in total
cholesterol (134 to 170 mg/dL, P , 0.01), LDL-c (80 to 120 mg/dL, P , 0.05), Triglycerides (124 to 162
mg/dL, P , 0.01) and VLDL-c (25 to 32 mg/dL, P , 0.01) when compared to the untreated patients. HDL-
c showed significant increase from baseline to 6 months (32 to 51 mg/dL), but there was a significant
drop at 12 months (51 to 44 mg/dL) in patients on HAART.
Conclusions: Significant lipid and lipoprotein alterations are seen after 48 weeks of HAART. Serum
lipid profile should be monitored annually to avoid morbidity among patients on generic HAART.
257 Does toxicity to fixed dose stavudine, lamivudine
and nevirapine regimen affect virologic suppression
among HIV infected adults at the Infectious Diseases
Institute, Makerere University?
Fred C. Semitala, 1 Harriet Mayanja- Kizza, 1 Andrew Kambugu, 2

Agnes K. Kiragga, 2 Barbara Castelnuovo, 2 Robert Kalyesubula, 1
Walter Schlech,3 Robert Colebunders,4 Keith McAdam,2 and Moses R. Kamya1
1
Department of Medicine Makerere University, Kampala, Uganda; 2Infectious Diseases Institute,
Makerere University, Kampala, Uganda; 3Dalhousie University Halifax, Canada; 4Department of Clinical
Sciences, Institute of Tropical ?Medicine Antwerp, Belgium
Introduction: Use of fixed dose combination of stavudine, lamivudine and nevirapine (Triomune)
for the treatment of HIV/AIDS is wide spread in Resource Limited Settings. However, Triomune has
been associated with toxicities which might affect the virologic outcome. The extent of the toxicities
and their effects are not well studied in Uganda.
Objectives: To establish factors associated with selected Triomune related toxicities and the effect of
toxicities on virologic outcome during the first year of treatment among HIV-1 infected adults at the IDI.
Methodology: Retrospective cohort study using the database and records from a cohort of
patients started on Triomune between April 2004 and April 2005. Univariate and multivariate analysis
was used to compare demographic, clinical and laboratory characteristics of patients who developed
toxicity with those who did not.
Results: 388 patients were included in this study; 73% were female. Median age was 37 (IQR = 32–
44) years, median BMI 20 kg/m2 (IQR = 18–22) and 92% were WHO stage 3 or 4, median CD4 count was
91 (IQR = 14–158) cells/mm3 and median viral load 5.4log10 [(IQR = 5.1–5.7)] copies /mL. 143 (37%)
patients developed at least one toxicity during the first year;. 122 (31%) peripheral neuropathy (PN),
16(4%) skin rash, 11 (3%) liver injury and 6 (2%) hyperlactaemia. Baseline factors associated with
toxicity were: over weight OR 5.69 (95% CI, 0.021–1.00), haemoglobin ,11.5g/dL [OR 1.92 (95% CI:
1.25–2.96)] and being started on 40 mg of stavudine compared to 30 mg [OR 1.96 (95% CI: 1.24–3.49)].
CD4 count .200 cells/mm [OR 2.39 (95% CI 1.18–4.84)] and viral load .5 log10 copies OR 1.82 (95% CI,
1.01–3.26), were associated with PN, while baseline CD4 count ,100 cells/mL [OR 3.27 (95% CI 1.60–
6.68)] with liver injury. Of the 306, patients with complete data at 12 months, 86.6% had virologic
suppression. By intention to treat, only 56.6% had viral suppression at 12 months and this was
statistically significant OR 5.43 (95% CI 3.09–9.63).
Conclusions: There was a high incidence of toxicity especially PN and toxicity was associated with
virologic failure. Over weight, low haemoglobin and receiving 40mg of stavudine were risk factors for
Triomune toxicity.
258 Intra patient evolution of Human Immunodeficiency
Virus Type 1 protease and reverse transcriptase in
antiretroviral naı̈ve patients
Uma Shanmugasundaram,1 Ku marasamy Nagalingeswaran,1

Kailapuri G. Murugavel,1 Saravanan Shanmugam,1 Vidya Madhavan,1
Sunil S. Solomon, 1 Suniti Solomon, 1 Kenneth H. Mayer, 2 and
Balakrishnan Pachamuthu1
1
Y.R.Gaitonde Centre for AIDS Research and Education, Chennai, India; 2Department of Medicine,
Brown University/Miriam Hospital, Providence, RI, USA
Objective: Intra patient HIV-1 evolution reflects the strong selection pressure driving viral escape
from cytotoxic T-lymphocyte (CTL) recognition. We studied the intra host variation of HIV-1 protease
(PR) and reverse transcriptase (RT) sequences from the antiretroviral na¨ve (ART) patients.
Methods: Peripheral blood samples were collected longitudinally between 10–12 months from
18 HIV-1 infected ART na¨ve patients. HIV-1 RNA was isolated using Qiagen extraction kit. RT (n = 18) and
PR (n = 10) gene was successfully amplified using nested PCR and direct sequencing was performed.
Intrapatient nucleotide and amino acid distance of RT and PR based on pairwise comparison was
performed using MEGA v3.1. The ratio of frequencies of synonymous mutations per nonsynonymous
site (ds/dn) was implemented using Synscan (www.hivdb.stanford.edu). Mann Whitney U Test applied
to compare the ds/dn ratio between PR and RT gene.
Results: All the sequences were HIV-1 subtype C. The median intrapatient nucleotide distance of
PR and RT measured based on pairwise comparison was 0.01 and 0.00 respectively. The majority of
nucleotide changes in the PR and RT gene involved synonymous substitutions, resulting in a ratio of
ds/dn .1 and the ratio was significantly lower (P = 0.001) in the PR sequences compared to RT
indicating that the former is less conserved than the latter. The amino acids under selection located
away from the active sites and some of which were under the predefined CTL epitope were observed
at frequencies of above 50%.
Conclusion: Negative selection observed in HIV-1 PR and RT gene indicates the absence of host’s
selective forces in the absence of antiretroviral therapy and the fixation of an escape variant in the
population could result in the loss of the epitope as a potential target to the immune system.
259 Quality of life among end-of-life former commercial
plasma donors infected with HIV and the care model
in rural Henan, China
Yu Sheng
Beijing P. R. China
Objective: To investigate the quality of life of rural, terminally ill former plasma donors, and to
discuss the trial hospice care provided by the Henan provincial government.
Method: A descriptive cross-sectional design was used. The 100 terminally ill AIDS participants in
this study were selected from twelve villages in Weishi county, Zhenping county and Tanghe County in
Henan province using convenience sampling. The Would Health Organization Quality of Life for HIV
(WHOQOL-HIV) BREF (WHO QOL-HIV-BREF Chinese version) was used to measure the quality of life of
the patients. The Memorial University of Newfoundland Scale of Happiness (MUNSH) was used to
measure the patients’ emotional feelings. Qualitative interviews and a focus group discussion were
used to investigate the hospice care model.
Results: Patients quality of the life score was 12.37 6 1.46 which is classified as moderate. The
patients scored higher in spirit/religion/self-faith a dimension and psychological area while in
independence dimension, social relationship dimension and environment dimension the scores are
relatively low but are all in the moderate level too. The average score of the patients’ feelings of
welfare is 20.91, which is also in the moderate level. The result which interviewed with village doctors
indicated that there was a lower stigma for the AIDS patients in that area because the most patients
were infected by plasma donation. On the other hand, they were depended on a very strong local
social system which consisted of village doctors, nurses and patient’s assistants to provide the care for
the end of life AIDS patients.
Conclusions: There are a modirate quality of life and feelings of welfare for end-of-life AIDS patients
in Henan. This care model which was base on local cultural social system is effective and available for the
end of life AIDS patients in Henan. It’s necessary to develop the nurses’ function and involve them into
the team for the HIV/AIDS treatment and prevention.
260 Scaling Up PMTCT Access Using a Hub and Spoke
Model in Nigeria, Sub-Sahara Africa
Dr. Akinmurele Timothy

Issues: PMTCT services in Nigeria were initially limited to comprehensive ART centers, limiting access.
Extending PMTCT to small secondary hospitals and primary health centers necessitates access to CD4
count and clinical staging for HIV+ women, linkage to an ART center for women requiring HAART for their
own health care, and quality site monitoring.
Description: PMTCT networks were developed regionally, with small secondary hospital and
primary health center ‘‘Spoke’’ sites linked to a larger hospital ‘‘Hub’’ sites. For HIV+ women at ‘‘Spoke’’
sites, blood was drawn for CD4 count and transported to the ‘‘Hub’’ site laboratory. Those HIV+ women
at ‘‘Spoke’’ sites requiring HAART for their own health care were provided transportation to the ‘‘Hub’’
site, while all other HIV+ women were offered short course ARV prophylaxis at the ‘‘Spoke’’ site.
A network coordinator from the ‘‘Hub’’ site provided training and site monitoring and organized
monthly network coordination meetings.
Results and Lessons Learned: Networks consisting of 3 ’Hub" sites and 13 ‘‘Spoke’’ sites providing
services in 2007 were evaluated. At ‘‘Hub’’ sites, 6,882 new ANC clients were served, with 74%
counseled, HIV tested and receiving results. At ‘‘Spoke’’ sites, 33,119 new ANC clients were served, with
87% counseled, tested and receiving results. 583 HIV+ pregnant women were identified/referred and
received services at the ‘‘Hub’’ sites, while 536 were identified and received services at the ‘‘Spoke’’ sites.
Women were less likely to return for delivery at the ‘‘Spoke’’ sites (48.5%) than ‘‘Hub’’ sites (61.2%)
Next Steps: The network model provided a fourfold increase in women accessing PMTCT services.
Use of a regional coordinator ensures quality of care and linkage and addresses implementation/
coordination challenges. Intensive interventions to reduce loss to follow up of HIV+ women are even
more critical as PMTCT services are provided at the primary health center level.
261 Impact of Community- and Home-Based Care on
HIV/AIDS Patients and Their Families
Emily Hauwa Umaru

Background: In efforts to mitigate HIV pandemic, the federal government of Nigeria collaborated
with PEPFAR implementers in selected health facilities around the federation to enhance services
provided to HIV infected and affected clients. IHVN-ACTION program through its multidisciplinary
team is building capacity for provision of comprehensive HIV treatment, care and support. This
includes HCT, Lab investigations, treatment/ adherence support, palliative care (care & support and
home based care). In addition Home based care services were commenced to enhance continuum of
care for PLHAs and infected children. Formal and informal care givers receive training in home care and
other community services to promote, restore and maintain a person’s maximum level of comfort,
function and health. Health professionals work in collaboration with volunteers to provide them with
technical assistance in their communities while volunteers in turn refer clients to health facilities.
Approach: Advocacy to policy and community leaders on HIV/AIDS is paramount to gain their
support for a successful program implementation. Community leaders are sensitized and intimated on
the benefits of the program and requested to nominate volunteers to support the program. In
addition volunteers are drawn PLHA support groups who also serve as peer educators to encourage
other PLWHAs. Health care providers (nurse coordinator and social workers) from the facility serve as
a link to provide technical assistance to the volunteers. These volunteers undergo six days intensive
training to equip them with knowledge and skills in HIV/AIDS care. Volunteers are trained to conduct
various activities such as home visit to the client for: Palliative/basic nursing care, psychosocial,
nutritional, spiritual and adherence support. They also assist with activities of daily living, provide HBC
kits where necessary, educate relatives and friends on care for PLHA, identify care givers for OVCs,
counsel and refer relatives and friends for HCT and psychosocial support. Networking, linkages and
collaboration with other CBOs/NGOs are done to leverage client support for other services. They
conduct ongoing community sensitization for HIV/AIDS prevention, stigma reduction and behavior
change for positive living, follow up defaulters (loss to follow up/poor adherence) and refer clients for
health services, mobilize communities for ongoing mobile HCT in collaboration with counselors/Lab
teams.
The HBC services had a humble beginning, with only a few clients as many were afraid they’ll be
stigmatized by neighbors and other family members. This number has grown tremendously, services
have continued to expand as volunteers now identify more patients in need of care through

261 Impact of Community-and Home-ased Care on HIV/
AIDS Patients and Their Families?
(continued)
aggressive community mobilization, serve as buddy for community adherence and refer sick persons
to hospital for HCT etc. To enhance their impact the HBC nurses were trained by ACTION to do HIV
counseling and testing in the community and at homes. The linkage between the community and the
hospital has been strengthened through the services of the HBC team. The utilization of volunteers has
made it possible for smooth transition and effective provision of palliative care from the facility into
the community thus enhancing the proverbial adage that ‘‘human beings are their brothers’ keepers’’.
The community involvement and support has been overwhelming to the surprise of both clients and
care providers. They have mobilized additional resources e.g. funds and food items to assist clients.
Radio programs and other modes of communication are used to create awareness about HIV and
services for those infected and affected. Ongoing HIV/AIDS sensitization to communities and in the
media is gradually changing the attitude of individuals. Family and community members are informed
on HIV/AIDS and provide care for their loved ones which has tremendously reduced stigma, people
can now talk about HIV, and opt for HCT, treatment and care.
Challenges: Despite the effort from CHBC volunteers, some challenges abound, like need for
increased funding support and transportation assistance especially to hard to reach areas as their
client load grows.
Conclusion: Despite challenges, the benefits of community and home based services have been
encouraging to the program and service providers. Many volunteers verbalize how improving patient
quality of life and ability for self care continue to keep them motivated. Many of the patients
themselves return to work as volunteers and adherence supporters for others so as to share their lives
with others.
262 Prevalence of Norovirus infections in children from
Iquitos, Peru
Daniel Velasquez Portocarrero

Jr. Enrique Barron, PERU
Background: Noroviruses are the most commonly identified cause of outbreaks and sporadic cases
of acute Gastroenteritis. Norovirus is a group of non-enveloped RNA viruses that belong to the family
Caliciviridae. A number of studies in both developed and developing countries have shown that
Norovirus is a major enteric pathogen, (second only to rotavirus), causing acute diarrhea in children which
can sometimes be fatal.
Objective: To characterize the burden of diarrheal disease caused by Norovirus in children from
Iquitos, Peru.
Methods: Norovirus testing was done for genogroup I and genogroup II using sensitive and
specific TaqMan Real-time Reverse Transcription-PCR in 338 fecal specimens from children in a peri-
urban area near Iquitos, Peru.
Results: Norovirus was present in 23.6% of 254 diarrheal samples as compared to 9.3% of 84 non-
diarrheic controls (P = 0.007). GII was present in 14.8% of diarrheal stools tested and 5.3% of stools
from children in the absence of diarrhea. GI was present in 8.7% of diarrheal stools and 4.0% of
asymptomatic controls. The presence of GII, but not GI was associated with the presence of diarrhea
(Fisher’s exact 0.03 and 0.22, respectively) although the power of the study to detect such a difference
with this sample size was only 17%. We also found that the identification of Norovirus in stool samples
was age-related and decreased 32% per year (95% CI: 15–45%). The characterization of the enteric
disease caused by Norovirus was done by comparing the presence of clinical symptoms and signs.
Norovirus disease detected by active surveillance is best characterized as acute uncomplicated watery
diarrhea. There was no difference in maximum stool frequency reported in a 24 hour period during the
diarrheal episode in Norovirus as compared with diarrhea from other causes (6.75 and 7.03
episodes/24 hrs, P = 0.58). There was no difference in the reported incidence of nausea, vomiting, or
fever between Norovirus and all-cause diarrhea.
Conclusions: This study demonstrates that Norovirus is an important enteric pathogen in this
population. GII was more prevalent than GI, which is consistent with other studies of Norovirus in the
world. It also documents for the first time the prevalence of Norovirus infections in Peru.
263 Adherence to highly active antiretroviral treatment
and related factors in drug users with HIV/AIDS
Wang Honghong
School of Nursing, Central South University, Changsha, China
Objectives: To explore adherence to antiretroviral treatment (ART) and related factors in drug
users with HIV/AIDS.
Methods: From July to September 2007, 111 HIV-infected drug users who received national free
HAART were investigated in the HAART clinics in Hengyang, Yueyang, and Chenzhou districts of Hunan
Province. A questionnaire of Community Programs for Clinical Research on AIDS (CPCRA )
Antiretroviral Medication Self-Report was administered to assess adherence to ART, and Zung
Depression Scale and Adaptation Partnership Growth Affection Resolve Scale were used to assess
patients’ depression and family function respectively.
Results: The average level of adherence to ART was 83%. Among 111 patients, 28.8% of patients
reported poor adherence and took medication less than 90%. There were 83.9% patients
demonstrating depressive symptom and only 30.6% patients’ family had good function. Logistic
regression analysis showed that the degree of depression and treatment time were significantly
associated with adherence negatively, while family function and the time of being free from drug were
positively associated with adherence.
Conclusion: The level of adherence to ART is low in the drug users with HIV/AIDS. Comprehensive
interventions are needed to improve adherence to ART, including managing depression, encouraging
drug rehabilitation, improving family function, and evaluating adherence periodically.
264 Immunologic recovery among HIV infected children
on first line antiretroviral therapy in Kano, Nigeria
Bashir M. Zubayr,
Background: HIV disease progression in infants and children is rapid compare to adults. This picture
is gloomier in sub Sahara Africa due to the co-morbidities of malaria, diarrhea disease and malnutrition.
Therefore, the goal of treatment and care of the HIV infected child is to promote health and prevent
disease progression. This present study evaluates immunologic recovery after one year of antiretroviral
therapy using CD 4 cells counts.
Aims and Objectives: To determine the pattern of CD4 count rise after one year of Zodovudine
base first line therapy in HIV infected children.
Method: HIV infected children who were treatment na¨ve and commenced on Zodovudine based
regimen were enrolled into the study. Age, weight, CD4 counts and Hematocrit prior to initiation of
treatment were all recorded and entered into a proforma. These parameters were reassessed again at 6
and 12 months on treatment and recorded.
Data collected was analyzed using SPSS version 12 and presented as means and standard
deviation.
Results: A total of 92 HIV infected children completed the study, of which 48 were males and 44
females. Mean age at initiation of therapy was 5.4years while mean baseline Cd4 counts was 688
cell/mm. mean CD4 counts at 12 months was 746 cells, gain in CD4 among subjects was found to be
statistically significant. (P , 0.001)
Conclusions: There is significant increase in the Cd4 count after one year of treatment but no
significant change in weight and hematocrit.

Adeyemi 2009

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11th Anniversary Annual International

Meeting of the Institute of Human Virology

Communications and Press Policy

ONLINE ABSTRACTS TITLE INDEX

102 Visualization of HIV-1 entry pathways in cells

104 A V3-independent pathway to resistance to small molecule CCR5 inhibitors

HIV Infection Early Events

107 Escape from Neutralizing Antibody in Early Subtype C HIV-1 Infection

116 Molecular Studies of HIV-1 Rev Function

117 CA-Dependent HIV-1 Nuclear Import Relies on Transportin-SR2

118 Self-Inactivation of HIV by its own RT/Rnase H

123 AIDS Restriction Genes-Genome Wide Association Study

124 NK cells: good guys or bad guys?

126 Host genetic determinants of response to HIV-1: A genome-wide perspective

130 Inhibition of HIV-1 Release by Cell Permeable Peptides

137 Presence and Role of HLA-C in HIV-1 Infection

140 A Retrospective Study of HIV-Exposed Infacts/Children in Kitwe, Zambia

Friday, September 12, 2008

146 a4b7: A Newly Discovered Receptor for HIV

147 Hepatitis C and cancer

148 Towards a genetics of cancer resistance

150 HIV and cancer: current trends and insights

151 MicroRNAs and the biology of KSHV infection

154 Apobec3: A Modern Twist to A Classic Retroviral Mystery

157 Vpr as a mediator of proteasomal degradation and other functions

158 The SET complex acts as a barrier to autointegration of HIV-1

160 Polarization of M1 and M2 macrophages and HIV infection

161 Peptide stabilization of gp120 confirmation: a novel vaccine candidate

Saturday, September 13, 2008

174 The C5 region of gp120; a therapeutic vaccine target?

176 Vector-based Vaccines for Cancer Therapy

179 Rational Vaccine Design and the Development of an AIDS Vaccine

182 A Novel Model for In Vivo SIV Neutralization

183 Morphogenomic immune responsiveness to preventive/therapeutic vaccines

188 Targeted delivery of antiHIV siRNAs to T cells in vivo

189 Defective HIV-1 Proviral Genomes in Natural Viral Suppressors

190 The Immune Response to HIV: Friend or Foe

193 The HIV-Positive Inpatient: Psychosocial Risks and Adherence Implications

193a Psychosocial Contributors to Antiretroviral Adherence: Stability and Change

195 Stimulation of reverse cholesterol transport potently suppresses HIV-1 replication

200 Design, Synthesis, Anti-HIV and Cytotoxicity of Novel Heterocyclic Compounds

200a Inhibition of HIV replication and integrase activity by isatin derivatives

201 Pathogenesis induced acidosis and hpercalceamia evident in HIV/AIDS disease

209 TNF-a is an autocrine factor for Va2Va2 T cell

216 Genetics of Fc Gamma Receptors IIa and IIIa in Tanzania

FOGARTY SCHOLARSHIP ABSTRACTS

235 Quality ART Scale Up Through Regionalization of Laboratory Services In Nigeria

240 Patient Retention in a University Hospital-based ART program in Uganda

245 Outcome of early infant diagnosis in exposed infants in Northern Nigeria

262 Prevalence of Norovirus infections in children from Iquitos, Peru

Carole A. Bewley,1 Son N. Lam,1 Chih-Chin Huang,2 Priyamvada Acharya,2

Dr. Kleio Anastasopoulou

Laurel A. Lagenaur,1,2 Vadim A. Villarroel,1 and Edward A. Berger1

Ronald S. Veazey1 and Thomas Hope2

George M. Shaw, MD, PhD

Background: Identification of complete (9kb) genomes of HIV-1 viruses responsible for

Reinhard Kurth, Nadine Beimforde, Kirsten Hanke, and Norbert Bannert

Mark K. Lafferty, Lingling Sun, Wuyuan Lu, and Alfredo

Dr. Marie-Lise GOUGEON

Janet L. Douglas, Kasinath Viswanathan, Matthew N. McCarroll, Jean K. Gustin,

L. Gu, V. W. Gautier, N. Sheehy, T. Tsuji, H. Hayakawa, and W. W. Hall

Kyeongeun Lee, Thomas D. Martin, Zandrea Ambrose, Alok Mulky, and

K. Moelling, A. Matskevich, L. Wittmer-Elzaouk, J. Heinrich, J.-S. Jung,T. Kwok,

Placebo (n = 209) MVC QD (n = 414) MVC BID (n = 426)