In vivo analysis of HIV replication and persistence in the myeloid compartment
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1 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
2 BLT Mouse Dendritic Cell CD8+ CD4+
3 Macrophages/Microglia and HIV HIV-1 has evolved the ability to infect nondividing 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. Verani et al Mol. Imm. 2005
4 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.
5 Experimental Design: Making MoMs CD34 + Cells
6 Systemic human reconstitution in MoM 44% 47% 6.1% 28% 36% 35% 87% 62% <1% <1% <1% <1%
7 Reconstitution of MoM brain with human myeloid cells
8 Are MoM susceptible to HIV-1 infection?
9 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, CH env, JR-CSF, BaL, CH058, RHPA, HIV-2 (7321A), ADA or THRO CH env contains a macrophage-tropic envelope isolated from a patient
10 Viruses evaluated in MoM, BLT, & TOM as demonstrated by plasma VL (& tissue vdna) JR- CSF CH040 CH env ADA BaL CH058 RHPA THRO HIV-2 (7321A) MoM 0/3 0/1 high 14/14 16/16 1/3 1/1 high 0/4 0/1 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 mouse TOM=T cell only humanized mice BLT=bone marrow, liver, thymus humanized mouse nd= not done
11 Sustained replication of virus in MoM CH040 (n=9) BLT TOM
12 Sustained replication of virus in MoM CH env (n=11) BLT TOM
13 Sustained replication of virus in MoM ADA (n=2) BL T
14 Lack of detectable levels of vdna in the peripheral blood cells from MoM
15 Systemic presence of HIV in MoMs (tissue vdna)
16 Systemic presence of HIV in MoMs (tissue vrna)
17 HIV infected cells are present in MoM brain hcd45 hcd68 HIV p24 Dr. Angela Wahl
18 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 CH CH N N
19 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, CH , and ADA) Replication competent virus can be isolated from all infected MoM tissues
20 ART regimen Raltegravir integrase inhibitor Tenofovir NRTI Emtricitabine NRTI Mice administered treatment daily i.p.
21 ART effectively suppressed VL in CH040 infected MoMs
22 ART effectively suppressed VL in CH env infected MoMs
23 Viral load is effectively suppressed using ART in HIV infected MoM
24 Differences in suppression kinetics between MoMs, ToMs and BLTs Hu Type MoM BLT ToM MoM BLT Virus CH env CH040 Weeks to undetectable plasma viral load 1-2 weeks 3-6 weeks 2-7 weeks 1-3 weeks 3-7 weeks
25 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.
26 Jenna Honeycutt Angela Wahl Perry Tsai Rae Ann Spagnuolo Carolina Baker Patricia Sheridan Glenn Matsushima George Shaw Ron Swanstrom David Margolis Acknowledgements
27 Questions?
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