4 years ago

Identification of Genetically Intact HIV-1 Proviruses in Specific CD4+ T Cells from Effectively Treated Participants

Identification of Genetically Intact HIV-1 Proviruses in Specific CD4+ T Cells from Effectively Treated Participants
Bonnie Hiener, Lina Odevall, Sarah Palmer, Susanne von Stockenstrom, Timothy E. Schlub, Daniel Douek, Frederick M. Hecht, Eli A. Boritz, Teri Liegler, Nicolas Chomont, Jeffrey M. Milush, Rémi Fromentin, Kirston Barton, Bethany A. Horsburgh, Steven G. Deeks, Elizabeth Sinclair, John-Sebastian Eden, Rebecca Hoh, Eunok Lee


Latent replication-competent HIV-1 persists in individuals on long-term antiretroviral therapy (ART). We developed the Full-Length Individual Proviral Sequencing (FLIPS) assay to determine the distribution of latent replication-competent HIV-1 within memory CD4+ T cell subsets in six individuals on long-term ART. FLIPS is an efficient, high-throughput assay that amplifies and sequences near full-length (∼9 kb) HIV-1 proviral genomes and determines potential replication competency through genetic characterization. FLIPS provides a genome-scale perspective that addresses the limitations of other methods that also genetically characterize the latent reservoir. Using FLIPS, we identified 5% of proviruses as intact and potentially replication competent. Intact proviruses were unequally distributed between T cell subsets, with effector memory cells containing the largest proportion of genetically intact HIV-1 proviruses. We identified multiple identical intact proviruses, suggesting a role for cellular proliferation in the maintenance of the latent HIV-1 reservoir.

Publisher URL: http://www.cell.com/cell-reports/fulltext/S2211-1247(17)31386-4

DOI: 10.1016/j.celrep.2017.09.081

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