Brain microglia serve as a persistent HIV reservoir despite durable antiretroviral therapy
Yuyang Tang, … , David M. Margolis, Guochun Jiang
Yuyang Tang, … , David M. Margolis, Guochun Jiang
Published June 15, 2023
Citation Information: J Clin Invest. 2023;133(12):e167417. https://doi.org/10.1172/JCI167417.
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Research Article AIDS/HIV

Brain microglia serve as a persistent HIV reservoir despite durable antiretroviral therapy

Abstract

Brain microglia (MG) may serve as a human immunodeficiency virus 1 (HIV) reservoir and ignite rebound viremia following cessation of antiretroviral therapy (ART), but they have yet to be proven to harbor replication-competent HIV. Here, we isolated brain myeloid cells (BrMCs) from nonhuman primates and rapid autopsy of people with HIV (PWH) on ART and sought evidence of persistent viral infection. BrMCs predominantly displayed microglial markers, in which up to 99.9% of the BrMCs were TMEM119+ MG. Total and integrated SIV or HIV DNA was detectable in the MG, with low levels of cell-associated viral RNA. Provirus in MG was highly sensitive to epigenetic inhibition. Outgrowth virus from parietal cortex MG in an individual with HIV productively infected both MG and PBMCs. This inducible, replication-competent virus and virus from basal ganglia proviral DNA were closely related but highly divergent from variants in peripheral compartments. Phenotyping studies characterized brain-derived virus as macrophage tropic based on the ability of the virus to infect cells expressing low levels of CD4. The lack of genetic diversity in virus from the brain suggests that this macrophage-tropic lineage quickly colonized brain regions. These data demonstrate that MG harbor replication-competent HIV and serve as a persistent reservoir in the brain.

Authors

Yuyang Tang, Antoine Chaillon, Sara Gianella, Lilly M. Wong, Dajiang Li, Theresa L. Simermeyer, Magali Porrachia, Caroline Ignacio, Brendon Woodworth, Daniel Zhong, Jiayi Du, Eduardo de la Parra Polina, Jennifer Kirchherr, Brigitte Allard, Matthew L. Clohosey, Matt Moeser, Amy L. Sondgeroth, Gregory D. Whitehill, Vidisha Singh, Amir Dashti, Davey M. Smith, Joseph J. Eron, Katherine J. Bar, Ann Chahroudi, Sarah B. Joseph, Nancie M. Archin, David M. Margolis, Guochun Jiang

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Figure 5

HIV was outgrown from brain-derived MG.

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HIV was outgrown from brain-derived MG. (A) Cell-free HIV RNA (gag) in t...
(A) Cell-free HIV RNA (gag) in the culture supernatants was measured in the SAHA- and CM272-treated MG cultures (105 cells/well) isolated from the parietal cortex of donor 2 (n = 3). (B) Outgrowth HIV was tracked over time by measuring viral RNA release in the culture supernatant of SAHA- and CM272-treated MG wells (from the indicated brain region of donor 2) after addition of CD8-depleted PHA PBMC blasts (n = 3). (C) MG QVOA and de novo infection by human brain MG–derived HIV. After MG were isolated from the brain of PWH, cells were plated in the 24-well plates with limited dilutions and cultured for 14 days in the presence of ART, allowing the cells to settle down and attach to the surface. The latent HIV was activated with SAHA and CM272 for another 7 days, and then the LRAs were washed out. For the MG QVOA, LRA-treated MG were cultured with CD8-depleted, HIV– PBMC PHA blasts or MOLT-4/CCR5 cells. Viral outgrowth was measured on day 21 and was further confirmed on day 28. De novo HIV infection was used to assess MG-derived, replication-competent HIV via inoculation of virus from LRA-stimulated MG culture into MG or PBMC blasts isolated from HIV– donors. HIV replication was assayed by HIV RNA and p24 released into culture supernatants. ART consisted of raltegravir plus darunavir plus nevirapine. (D and E) The IUPM of MG and CNS T cells was calculated by standard viral outgrowth assay (measuring HIV p24 antigen release in the wells) (D) and by RT-ddPCR to measure viral RNA+ wells (E) (n = 4).