Journal
CELL REPORTS
Volume 21, Issue 3, Pages 813-822Publisher
CELL PRESS
DOI: 10.1016/j.celrep.2017.09.081
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Funding
- Delaney AIDS Research Enterprise (DARE) [1U19AI096109, 1UM1AI126611-01]
- Foundation for AIDS Research [amfAR 108074-50-RGRL]
- Australian Centre for HIV and Hepatitis Virology Research (ACH2) [2015-43]
- UCSF-GIVI Center for AIDS Research [P30 AI027763]
- Australian National Health and Medical Research Council [AAP1061681]
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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 (similar to 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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