| Publication Abstract Display | | Type: Published Abstract | | Title: Presence of Intact HIV DNA Variants in the Brain and Lymphoid Tissues during ART. | | Authors: Oliveira M, Murrell B, Vitomirov A, Vollbrecht T, Kumar V , Kumar N, Gouaux B, Moore D, Jarmush A, Vargas F, Dorrestein D, Ellis R, Smith D, Gianella S | | Year: | | Publication: CROI | | Volume: Issue: Pages: | | Abstract:Although antiretroviral therapy (ART) reduces HIV RNA below the detection limit in blood plasma, HIV reservoirs persist in cellular compartments. Here, we characterize the size and composition of the HIV DNA reservoirs in brain and lymphoid tissues.
We evaluated post-mortem brain and peripheral lymphoid tissues from 12 persons living with HIV (PLWH) obtained from the National NeuroAIDS Tissue Consortium. All donors died between 2001-2014, with virologic suppression on ART (<50 or 400 copies/ml, assay-dependent), and without evidence of central nervous system opportunistic disease. Presence of ART in post-mortem brain was confirmed by mass spectrometry. Total DNA was extracted from each tissue sample and levels of HIV DNA (gag) were quantified by droplet-digital PCR. The genotypic composition of the HIV DNA populations was evaluated by high-throughput single genome amplification using the PacBio platform to sequence Full-length HIV envelope (FL HIV-env).
We evaluated post-mortem tissues from 9 men and 3 women with a median age of 52 years (range: 40-66). Donors were on ART at the last visit, which occurred a median of 3 months prior to death (range: 1-4). Presence of Tenofovir or Lamivudine was confirmed in 6 out of 8 donors’ brain tissues by mass spectrometry. All donors had detectable HIV DNA in brain (frontal [FC] or occipital [OCC] cortex) and lymphoid tissues (lymph node [LN] or spleen [SP]). A total of 180 individual FL HIV-env sequences were obtained across donors (FC, n= 7 donors; OCC, n=3; LN, n=3; SP, n=8). For 10 donors, FL HIV-env sequences were obtained from paired brain and lymphoid tissues. Maximum likelihood phylogeny (figure) suggests that HIV compartmentalization patterns differ between donors, with four donors showing evidence of HIV DNA compartmentalization (p<0.05). Overall, 143 FL HIV-env sequences were genetically intact, while 37 sequences were non-functional, with major deletions, frameshifts and stop codon mutations (figure). For one donor, we found 23 clonal sequences with a frameshift mutation that was present in both brain and spleen, suggesting migration of cells with clonal provirus between tissue compartments.
HIV DNA was detected in brain and lymphoid tissues despite long-term ART. Most HIV DNA populations in brain and lymphoid tissues appeared to have intact env genes and were often compartmentalized from each other. Characterizing the composition of the HIV reservoirs in anatomic compartments is crucial for future HIV cure strategies. |
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