cell noncytotoxic antiviral response suppresses siv replication
TRANSCRIPT
CD8+ T-Cell Non-Cytotoxic Antiviral Response (CNAR) Suppresses SIV Replication
Meenakshi Vanka
Emory University, Class of 2013
Sacha Lab, Oregon National Primate Research Center
Overview Introduction to HIV/AIDS epidemicCD8+ T-cells and HIV infection CD8+ T-cell Non-Cytotoxic Antiviral Response
(CNAR)
HIV: A Global Epidemic
• CD4+ T-cells
• CD8+ T-cells
HIV infection
• SIV
• HIV
Course of infection
Matano et al. (1998), Jin et al. (1999), Schmitz et al. (1999), Metzner et al. (2000)
plas
ma
vira
l con
cent
ratio
n
Weeks Years
Role of CD8+ T-Cells H
IV-s
peci
fic C
D8
freq
uenc
y
plas
ma
vira
l con
cent
ratio
n
Matano et al. (1998), Jin et al. (1999), Schmitz et al. (1999), Metzner et al. (2000)
Weeks Years
Cytotoxic CD8+ T-cell function
CD4+ T-cellSIV
CD8+ T-cell
CD8+ T-Cell Non-Cytotoxic Antiviral Response (CNAR)
CD8+ T-cell suppression of viral replication without killing infected cells.
Mediated by soluble Cell Antiviral Factor (CAF).Does not depend on cell contact.Present in HIV infected individuals, but not naive
individuals. Positively correlated with an asymptomatic
clinical state.
Non-Cytotoxic CD8+ T-cell function
CD4+ T-cellSIV
CD8+ T-cell
CAF
ObjectivesTo optimize assay set up and test CNAR activity in
the rhesus macaque model. To better characterize protective CD8+ T-cell anti-SIV
activity.
HypothesisViremic controllers and SIV vaccinated (CMV
vector) animals will exhibit CNAR activity.
Methods1. Isolate lymphocytes from animal blood.
Activation
CD4+ T-cell CD8+ T-cell
2. Isolate CD4 T-cells and CD8 T-cells.
3. Activate CD4 and CD8 T-cells in vitro.
Methods (continued)
Add CD8+effectors
SIV
SIV-infected CD4+ T-cells (Targets)
CD4+ T-cells
Infect CD4+ T-cells with SIV.
Co-culture CD8+ Effectors and CD4+ Targets.
Stain for virus.
Cell Population Analysis
Gag p27
CD4
Uninfected 7 days Post Infection
.013 31.9
Viral Suppression
Gag p27
CD4
No CD8+ T-cells added CD8+ T-cells added
39.9 15.7
CNAR activity in viremic controllers
0
10
20
30
40
50
60
70
80
90
100
Viremic Controller 1 (22559)Viremic Controller 2 (22642)
Effector:Target Ratio
Perc
ent G
ag p
27 S
uppr
essio
n
.25:1 .5:1 1:1 2:1
CNAR activity in SIV vaccinated (CMV vector) animals
0
10
20
30
40
50
60
70
80
90
100
23716
24513
24552
Effector:Target Ratio
Perc
ent G
ag p
27 S
uppr
essio
n
.25:1 .5:1 1:1 2:1
CNAR activity in naïve animals
.25:1 .5:1 1.:12:1
0
10
20
30
40
50
60
70
80
90
Naïve Animal 1 (30673)Naïve Animal 2 (30674)
Effector:Target Ratio
Perc
ent G
ag p
27 S
uppr
essio
n
CNAR activity in MHC mismatched animals
0
10
20
30
40
50
60
70
80
90
100
22559 E/28401 T22559 E/27603 T22642 E/28401 T22642 E/27603 TViremic Controller 1 (22559)Viremic Controller 2 (22642)
Effector:Target Ratio
Perc
ent G
ag p
27 S
uppr
essio
n
.25:1 .5:1 1:1 2:1
Summary The CD8+ T-Cell Non-Cytotoxic Antiviral
Response (CNAR) best suppresses SIV replication in viremic controllers and SIV vaccinated (CMV vector)/SIV protected animals.
These findings reinforce CNAR activity seen in HIV infected individuals.
Future DirectionsPerform transwell assays to confirm that CNAR
is mediated by soluble Cell Antiviral Factor
(CAF). non-cytolytic in nature.
Screen more naïve animals to determine whether CNAR is an SIV specific response.
Acknowledgements
Ben Burwitz Jonah SachaJason ReedEnrique LeonSilver Family Foundation