in vivo analysis of hiv replication and persistence in the myeloid compartment
DESCRIPTION
In vivo analysis of HIV replication and persistence in the myeloid compartment. J. Honeycutt, A. Wahl, J. Foster, R.A. Spagnulo and J. Victor Garcia. Division of Infections Diseases UNC Center for AIDS Research University of North Carolina, at Chapel Hill. BLT Mouse. Dendritic Cell. - PowerPoint PPT PresentationTRANSCRIPT
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In vivo analysis of HIV replication and persistence in the myeloid
compartment
J. Honeycutt, A. Wahl, J. Foster, R.A. Spagnulo and J. Victor Garcia
Division of Infections Diseases UNC Center for AIDS Research
University of North Carolina, at Chapel Hill
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BLT Mouse
Dendritic Cell
CD8+CD4+
http://cnx.org/
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Macrophages/Microglia and HIV
Verani et al Mol. Imm. 2005
HIV-1 has evolved the ability to infect non-dividing macrophages.
HIV-1 infection and replication in microglia/ macrophages differs from that in CD4+ T lymphocytes.
Specific cellular factors are required for HIV-1 transcription in macrophages.
Monocytes/macrophages may serve to transmit HIV-1 to the central nervous system.
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MoM: Myeloid-(only) Mouse model
• Created by transplantation of human CD34+ stem cells into pre-conditioned (irradiated) adult NOD/SCID mice.
• Human cells differentiate into B and myeloid lineages.
• However, these mice do not support human T cell production.
• Therefore the only targets for HIV infection are myeloid cells.
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Experimental Design: Making MoMs
CD34+ Cells
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Systemic human reconstitution in MoM
36% 35% 87% 62%
44% 47% 6.1% 28%
<1% <1% <1% <1%
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Reconstitution of MoM brain with human myeloid cells
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Are MoM susceptible to HIV-1 infection?
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Experimental Design
• Viruses evaluated via intravenous injection – Typical dose contained 360,000 TCIU– “high” dose contained 720,000 TCIU of virus
• MoM injected with: CH040, CH0404013env, JR-CSF, BaL, CH058, RHPA, HIV-2 (7321A), ADA or THRO– CH0404013env contains a “macrophage-tropic”
envelope isolated from a patient
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Viruses evaluated in MoM, BLT, & TOM as demonstrated by plasma VL (& tissue vDNA)
JR-CSF CH040
CH040-4013 env
ADA BaL CH058 RHPA THRO HIV-2(7321A)
MoM0/30/1 high
14/14 16/16 1/3 1/1 high
0/40/1 high 0/3 0/3 0/3 0/2
BLT All All 6/6 1/1 1/1 1/1 All All 4/4
TOM All 4/4 4/4 nd nd nd nd nd nd
MoM=myeloid only humanized mouseTOM=T cell only humanized mice
BLT=bone marrow, liver, thymus humanized mousend= not done
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Sustained replication of virus in MoM CH040 (n=9)
BLT
TOM
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Sustained replication of virus in MoM CH0404013env (n=11)
BLT
TOM
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Sustained replication of virus in MoM ADA (n=2)
BLT
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Lack of detectable levels of vDNA in the peripheral blood cells from MoM
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Systemic presence of HIV in MoMs (tissue vDNA)
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Systemic presence of HIV in MoMs (tissue vRNA)
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HIV infected cells are present in MoM brain
hCD45 hCD68 HIV p24
Dr. Angela Wahl
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Rescue of replication competent HIV from tissues obtained from infected MoM
• Cells were plated and allowed to adhere overnight • Media was aspirated and 1 million CD4+ T cells (from healthy
donor) were added to the culture• HIV-RNA levels were measured in the supernatant after 10 days
Virus Weeks Infected ART? Liver Lung Spleen BM
ADA 6 N + + + +
CH040 4 N + + + +CH040-4013 5 N + + - +
CH040-4013 5 N + + + +
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HIV infection of MoMs
• MoM are systemically reconstituted with human myeloid and B cells (absence of T cells)
• MoM can systemically replicate HIV-1 over time Only certain viruses (CH040, CH040-4013, and ADA)
• Replication competent virus can be isolated from all infected MoM tissues
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ART regimen
• Raltegravir –integrase inhibitor
• Tenofovir –NRTI
• Emtricitabine –NRTI
Mice administered treatment daily i.p.
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ART effectively suppressed VL in CH040 infected MoMs
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ART effectively suppressed VL in CH0404013env infected MoMs
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Viral load is effectively suppressed using ART in
HIV infected MoM
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Differences in suppression kinetics between MoMs, ToMs and BLTs
Hu Type VirusWeeks to
undetectable plasma viral load
MoMCH040-
4013 env
1-2 weeks
BLT 3-6 weeks
ToM 2-7 weeks
MoMCH040
1-3 weeks
BLT 3-7 weeks
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Summary• MoM are systemically reconstituted with human
myeloid cells.• MoM infection with HIV results in sustained levels of
viral RNA in PB and tissues in the complete absence of T cells demonstrating the direct contribution of myeloid cells to HIV replication in vivo.
• HIV replication in MoM is efficiently suppressed by ART.
• In MoM myeloid cells repopulate the brain.• Myeloid cell in the brains of MoM are susceptible to
productive HIV infection.
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Jenna HoneycuttAngela WahlPerry TsaiRae Ann SpagnuoloCarolina Baker Patricia SheridanGlenn MatsushimaGeorge ShawRon SwanstromDavid Margolis
Acknowledgements
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Questions?