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Kanta Subbarao
Laboratory of Infectious Diseases, NIAID, NIH
NAS GoF symposium
December 15, 2014
Topics
Reverse Genetics
Generating viruses de novo
cDNA copy of genome
RNA
(+)
recombinant
virus
Biology of viruses
Virus Ecology
Viral Pathogenesis
Antiviral drugs
Immunoprophylaxis/immunotherapy
Vaccine development
Virus B
Not lethal
for mice
PB2x
PB1
PA
HA
NP
NA
M
NS
x
PB2
PB1
PA
HA
NP
NA
M
NS
PB2
PB1
PA
HAx
NP
NA
M
NS
Transmissibility
Indonesia/2005 H5N1 virus: Ron Fouchier (2012)
10 times
T156A
N158D
Q222L
G224S
Q226L
N224K
H103Y
T318I
Escape
Mutants
Cell monolayer X 2 h
+ Antibody/Antiviral
+ Antibody/Antiviral
4 days 37C
30 min 37C
OR
Experimental
animal model
Recover
virus
Virus
+ Antibody/Antiviral
Sequence to
identify
Escape
Mutants
Why?
To understand the biology of the virus
To map epitopes of antibodies
nsp3
nsp3
PL-protease
nsp5
nsp5
protease
Proteases
nsp12
nsp12
RdRp
nsp14 nsp16
nsp16
nsp14
Exonuclease 2O MT-ase
Polymerase
Proofreading
EM
Spike
Entry
Immune
Host Range
Evasion
Immunity
An3
Topics
Gain of Function
The term gain of function is a vague and unsatisfactory term
for microbiologists
One suggested alternative is aTRIP: an acronym for an
experiment that uses one or more of the DURC agents and
produces, aims to produce, or can be reasonably
anticipated to alter Transmission, Range and resistance,
Infectivity/immunity or Pathogenesis
Another suggestion is Gain of Virulence or Transmissibility
Comment/
Example
GoF?
Other
consequences:
LoF?
aTRIP/G
OVT?
Yes: usually
desired
no
Comment/
Example
Passage in
cell
culture/eggs
Serial
passage in
experimental
animals
SARS CoV
MA15
GoF?
Other
consequences:
LoF?
aTRIP/GO
VT?
Yes: usually
desired
Possible for
other cells
no
Possible:
often desired
Possible for
other species
RP/V
Comment/
Example
Passage in
cell
culture/eggs
GoF?
Other
consequences:
LoF?
aTRIP/GO
VT?
Yes: usually
desired
Serial
passage in
experimental
animals
SARS CoV
MA15
Possible:
often desired
Genetic
reassortment
of influenza
viruses
Influenza
vaccines GoF
yield, LoF
virulence
Yes; often
desired
Possible: often
desired
no
Comment/
Example
GoF?
Antiviral or
MAb resistant
mutants
Valuable
Yes: usually
information on desired
suitable
antiviral
strategies
LoF?
aTRIP/GO
VT?
Possible
reduction in
virulence and
fitness
no
Comment/
Example
Antiviral or
MAb resistant
mutants
Site directed
mutagenesis
GoF?
LoF?
aTRIP/GO
VT?
Valuable
Yes: usually
information on desired
suitable
antiviral
strategies
Possible
reduction in
virulence and
fitness
no
Used to
conclusively
prove the
molecular
basis of a
phenotype
Possible
No or
TRIP/VT
Possible
Topics
Are there viral targets that will not escape under immune pressure?
Measures
MERS
SARS
Non-HPAI
influenza
BSL3 with
respirators
Medical
surveillance
and support
Vaccines and
antivirals
N/A
N/A
Select agent
regulations
DURC
questionnaire
Priorities
Replace GOF with microbiological terms that precisely and
appropriately define the experiments of concern
MERS CoV
H5N2
H7N9
H10N8