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In vivo analysis of HIV replication and persistence in the myeloid compartment

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.

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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+ http://cnx.org/

  3. Macrophages/Microglia and HIV • 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. 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 28% 47% 44% 6.1% 35% 36% 87% 62% <1% <1% <1% <1%

  7. Reconstitution of MoMbrain 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, CH0404013env, JR-CSF, BaL, CH058, RHPA, HIV-2 (7321A), ADA or THRO • CH0404013envcontains a “macrophage-tropic” envelope isolated from a patient

  10. Viruses evaluated in MoM, BLT, & TOM as demonstrated by plasma VL (& tissue vDNA) 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 MoMCH040 (n=9) BLT TOM

  12. Sustained replication of virus in MoMCH0404013env(n=11) BLT TOM

  13. Sustained replication of virus in MoMADA (n=2) BLT

  14. Lack of detectable levels of vDNAin 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 hCD68 hCD45 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

  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, CH040-4013, 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 CH0404013envinfected MoMs

  23. Viral load is effectively suppressed using ART in HIV infected MoM

  24. Differences in suppression kinetics between MoMs, ToMs and BLTs

  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. Acknowledgements Jenna Honeycutt Angela Wahl Perry Tsai Rae Ann Spagnuolo Carolina Baker Patricia Sheridan Glenn Matsushima George Shaw Ron Swanstrom David Margolis

  27. Questions?

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