Viral Tissue Reservoirs Are Determined Early and Little Viral
RNA Is Detected during Suppression in the
Macaque ModelZandrea Ambrose, Ph.D.
Division of Infectious Diseases, University of Pittsburgh School of
Medicine, Pittsburgh, PA, USA
“Towards an HIV Cure” Pre-Conference Symposium20 & 21 July 2012
• infection, pathogenesis and immunity can be similar to that of HIV-infected humans
• procedures can be performed more easily and/or ethically than in humans• removal of vital organs at necropsy• acquisition of longitudinal samples • changing/testing novel treatment regimens
• model can be controlled in a way that humans cannot• known infecting virus, timing, and route of infection• known treatment adherence, past regimens, and timing of
treatments• scheduled blood and tissue sampling
Macaque Models of HIV/AIDS
HIV-1 Does Not Replicate Well in Monkeys
HIV-1
SIV
LTRLTR gag
PRenv
vif
vpx vpr
revtat nefRT IN
LTRLTR gagPR
env
vif
vpr vpu
revtat nefRT IN
RT-SHIV
LTRLTR gagPR
env
vif
vpx vpr
revtat nefINRT
Study Objectives
• Determine which tissues harbor viral DNA that could contribute to the reservoir• How early are these reservoirs established?• Are the levels different after suppressive therapy?
• Determine which tissues contain viral RNA• Are multiple sites contributing to viremia in untreated
animals?• Is there detectable viral RNA in the tissues of ART-
suppressed animals?
RT-SHIV Plasma Viremia
Therapy
1
2
3
4
5
6
7
0 5 10 15 20 25 30 35
Weeks post-challenge
Vir
al R
NA
co
pie
s/m
l p
lasm
a (l
og
)
PT8272
PT8030GN19
GG45GV08
GV40
TNV/FTC/EFV
TNV/FTC/EFV/L-870812
Therapy
No Therapy
1
10
100
1000
10000
10000 100000 1000000 10000000
Week 1 Plasma Viral Load
Lym
ph
oid
Tis
sue
vD
NA
(n
orm
aliz
ed t
o 1
06 C
CR
5)
0.996p < 0.0001
Week 1 Plasma Viremia Correlates with Lymphoid Viral DNA Levels
Little Viral RNA Detected in Tissues of Suppressed Animals
gag RNA copies/106 CD4 RNA copies in each tissue
PBMC Viral RNA Level is not Reflective of Plasma Viremia or Most Tissues
6760
8433
8232
GT29
GR6582
7280
30
GN19
GG45
GV08
GV401
10
100
1000
10000
100000
1000000
10000000PBMCPlasma
vRN
A (
per
10^
5 ce
lls o
r m
l pla
sma)
6760
8433
8232
GT29
GR6582
7280
30
GN19
GG45
GV08
GV401
10
100
1000
10000
100000
1000000PBMCPlasma
vRN
A (
per
10^
5 ce
lls o
r m
l pla
sma)
Untreated ART-Treated Untreated ART-Treated
Week 1/2 Week 28/30
• Viral DNA is detected in many tissues and PBMC of untreated and ART-treated macaques- < 4% of viral DNA from 2-LTR circles
• Levels of viral DNA in lymphoid tissues correlated with week 1 plasma viremia- reservoir size is established early after infection
• Tissue viral RNA levels were lower and less frequent in ART-treated macaques- No replication-competent virus was detected in LN resting
CD4+ T cells of ART-treated animals treated with 4 (< 0.5 IUPM)
• Viral RNA levels in PBMC were not consistent with treatment or plasma viremia
Conclusions
AcknowledgementsUniversity of Pittsburgh
Chris KlineJean NdjomouTamera Franks
John Mellors
AIDS & Cancer Virus Program, NCIJeff LifsonJake Estes
Mike Piatak, Jr.Jeremy Smedley
Vicky CoalterRebecca Kiser
MerckDaria HazudaJulio Quintero
Gilead SciencesNorbert Bischofberger
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