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Definitions
◼Originally derived from the Latin word ‘‘vacca’’
meaning cow
◼Pharmaceutical suspension or solution of an
immunogenic substance or compound(s) that is intended
to induce active immunity
◼Preparations containing antigenic substances that have
been shown to be capable of inducing a specific and
active immunity in man
VACCINES
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Principles of Vaccination
Immunity
• Self vs. non-self
• Protection from infectious disease
• Usually indicated by the presence of antibody
• Very specific to a single antigen
Principles of Vaccination
Active Immunity
• Protection produced by the person's own
immune system
• Usually permanent
Passive Immunity
• Protection transferred from another person
or animal as antibody
Principles of Vaccination
inject or administer
Antigen
• A live or inactivated substance (e.g., protein,
polysaccharide) capable of producing an
immune response
Antibody
• Protein molecules (immunoglobulin)
produced by B lymphocytes to help eliminate
an antigen
Passive Immunity
• Transfer of antibody from an exogenous
source
• Temporary protection
Sources of Passive Immunity
• Almost all blood or blood products
• Inactivated
kill the virus -> test can we make the vaccine or not
Inactivated Vaccines
Whole
we can prepare from:
• virus
• bacteria
Fractional
a component that include in inactivated vaccines
• protein-based
– subunit what is it? go home study it for exam
– toxoid some of them help you prevent disease
• polysaccharide-based some can cause immune response
(hypersensitivity) -> need to purify
– pure
carefully (phải high purity)
• Usually effective with one dose because long half-life and can
replicate
Live Attenuated Vaccines
• Severe reactions possible because replicate, they may become
dangerous again (become virulant
again)
• Unstable
Live Attenuated Vaccines
• Viral measles, mumps, rubella,
vaccinia, varicella, yellow
fever (oral polio) (rotavirus)
(influenza)
• pneumococcal
• meningococcal
• Haemophilus influenzae type b
Conjugate polysaccharide
• Haemophilus influenzae type b
• pneumococcal
Pure Polysaccharide Vaccines
• Not consistently immunogenic in
children <2 years of age
• No booster response
• Immunogenicity improved by
conjugation
VACCINES IN Immunization
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Mechanisms of VACCINES
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Vaccine preparation
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Traditional preparation
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Recombinant Vaccines
1. Subunit Vaccines
peptide vaccines
Genetic immunization
2. Attenuated Vaccines
3. Vector Vaccines
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VACCINES
Principle of Vaccination
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VACCINES
Subunit/Peptide Vaccines
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VACCINES
Subunit/Peptide Vaccines
• It has been showed that the capsid or envelope proteins are enough to
cause an immune response:
Herpes simplex virus envelop glycoprotein O.
Foot and mouth disease virus capsid protein (VP1)
Extracellular proteins produced by Mycobacterium tuberculosis.
• Subunit Vaccines
• Antibodies usually bind to surface proteins of the pathogen or proteins
generated after the disruption of the pathogen.
• Binding of antibodies to these proteins will stimulate an immune response.
• Therefore proteins can be use to stimulate an immune response.
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VACCINES
Subunit/Peptide Vaccines
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VACCINES
Subunit/Peptide Vaccines
CARRIER PROTEINS
Attenuated Vaccines
• Presently the cholera vaccine consist of a phenol-killed V. cholerae and it only last 3-6
months.
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VACCINES
Vector Vaccines
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VACCINES
Vector Vaccines
• A number of antigen genes have been inserted into the vaccinia virus
genome e.g.
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VACCINES
Vector Vaccines
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VACCINES
Bacterial Antigen Delivery Systems
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VACCINES
Vaccine Approval
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Vaccine Technology
Vaccine Technology
Vaccine Technology
Future Development scope of
vaccines
(a) Vaccine against Alzheimeir’s disease
(b) Vaccine for Meningitis C
(c) Super vaccine
(d) Immunomodulators
(e) Vaccination with Gas-Lighter
(f) Vaccine against cervical-cancer
(g) Vaccination without needles
VACCINE DESIGN
Polytope optimization
•Successful immunization can be obtained only if the
epitopes encoded by the polytope are correctly
processed and presented.
•The design of a polytope can be done in an effective
way by modifying the sequential order of the different
epitopes, and by inserting specific amino acids that will
favor optimal cleavage as linkers between the epitopes.
Polytope starting configuration
Polytope optimization Algorithm
• Optimization of four measures:
1. The number of poor C-terminal cleavage sites of
epitopes (predicted cleavage < 0.9)
2. The number of internal cleavage sites (within
epitope cleavages with a prediction larger than the
predicted C-terminal cleavage)
3. The number of new epitopes (number of
processed and presented epitopes in the fusing
regions spanning the epitopes)
4. The length of the linker region inserted between
epitopes.
Polytope final configuation
Prediction of antigens
• Protective antigens
• Functional definition (phenotype)
• Which antigens will be protective (genotype)?
• They must be recognized by the immune system
• Predict epitopes (include processing)
• CTL (MHC class I)
• Helper (MHC class II)
• Antibody
Function and conservation
• Some of the epitopes must exist in the wild type
• Conservation (http://www.ncbi.nlm.nih.gov/BLAST)
• Function
• When is it expressed?
• Where is it trafficked to?
• SecretomeP
Non-classical and leaderless secretion of eukaryotic proteins.
SignalP
Signal peptide and cleavage sites in gram+, gram-
and eukaryotic amino acid sequences.
TargetP
Subcellular location of proteins: mitochondrial,
chloroplastic, secretory pathway, or other.
• Expression level?
Selection of antigens
• Epitopes
• Polytope
• Proteins
• Helper epitopes
• Does it contain any
• Can they be added
• Hide epitopes
• Immunodominant and variable ones
Examples of antigen selections
Clustering of HLA alleles
Inside out:
1. Position in RNA
2. Translated regions (blue)
3. Observed variable spots
4. Predicted proteasomal cleavage
5. Predicted A1 epitopes
6. Predicted A*0204 epitopes
7. Predicted A*1101 epitopes
8. Predicted A24 epitopes
9. Predicted B7 epitopes
10. Predicted B27 epitopes
11. Predicted B44 epitopes
12. Predicted B58 epitopes
13. Predicted B62 epitopes
Strategy for the
quantitative ELISA assay
C. Sylvester-Hvid, et al., Tissue antigens, 2002: 59:251
b2m
Heavy chain Peptide-MHC
peptide Incubation complex
Development
+ peptide - peptide
Incubating in anti IFN- pre-coated plate for 18-20 h
•Coating Ab:
–Human IFN- MAb
(ENDOGEN, Pierce
Washing off the cells Biotechnology, Inc)
•Detection Ab:
–Human IFN- MAb, Biotin
Adding Biotin-anti IFN- labeled
–(ENDOGEN, Pierce
Biotechnology, Inc)
Adding a substrate
Automatical counting
Selected pathogens
• Web interface
http://www.imtech.res.in/raghava/propred
• Prediction Results
Prediction of Antibody epitopes
• Linear
– Hydrophilicity scales (average in ~7 window)
• Hoop and Woods (1981)
• Kyte and Doolittle (1982)
• Parker et al. (1986)
– Other scales & combinations
• Pellequer and van Regenmortel
• Alix
– New improved method (Pontoppidan et al. in preparation)
• http://www.cbs.dtu.dk/services/BepiPred/
• Discontinuous
– Protrusion (Novotny, Thornton, 1986)
Prediction of proteins structure
• Homology modeling
• Secondary structure prediction