[PDF][PDF] Identification of genetically intact HIV-1 proviruses in specific CD4+ T cells from effectively treated participants

B Hiener, BA Horsburgh, JS Eden, K Barton, TE Schlub… - Cell reports, 2017 - cell.com
B Hiener, BA Horsburgh, JS Eden, K Barton, TE Schlub, E Lee, S von Stockenstrom…
Cell reports, 2017cell.com
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 …
Summary
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.
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