title: Measuring the Frequency of Latent HIV-1 in Resting CD4⁺ T Cells Using a Limiting Dilution Coculture Assay.
abstract: Combination antiretroviral therapy (cART) can reduce HIV-1 viremia to clinically undetectable levels. However, replication competent virus persists in a long-lived latent reservoir in resting, memory CD4(+) T cells. The latent reservoir in resting CD4(+) T cells is the major barrier to curing HIV-1 infection. The recent case of the Berlin patient has suggested that it may be possible to cure HIV-1 infection in certain situations. As efforts to cure HIV-1 infection progress, it will become critical to measure the latent reservoir in patients participating in clinical trials of eradication strategies. Our laboratory has developed a limiting dilution virus outgrowth assay that can be used to demonstrate the presence and persistence of latent HIV-1 in patients. Here we describe both the original and a simplified version of the quantitative virus outgrowth assay (QVOA) to measure the frequency of latently infected resting CD4(+) T cells with replication competent provirus in patients on suppressive cART.
authors.affiliation: Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA.
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Laird
authors.affiliation: Department of Biomedical Informatics, Columbia University Medical Center, New York, NY, 10032, USA.
authors.forename: Daniel I S
authors.initials: DI
authors.lastname: Rosenbloom
authors.affiliation: Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA.
authors.forename: Jun
authors.initials: J
authors.lastname: Lai
authors.affiliation: Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA.|Howard Hughes Medical Institute, Baltimore, MD, 21205, USA.
authors.forename: Robert F
authors.initials: RF
authors.lastname: Siliciano
authors.affiliation: Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA. jsilicia@jhmi.edu.
authors.forename: Janet D
authors.initials: JD
authors.lastname: Siliciano
mesh_terms: D019380:Anti-HIV Agents / Q000627:therapeutic use*; D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D015658:HIV Infections / Q000188:drug therapy; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D014776:Virus Cultivation / Q000379:methods; D017735:Virus Latency / Q000187:drug effects
chemical_list: D019380:Anti-HIV Agents
keywords: HIV-1; Latency; Latent reservoir; Viral outgrowth assay