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Methods Mol Biol. 2016;1354:239-53. doi: 10.1007/978-1-4939-3046-3_16.

Measuring the Frequency of Latent HIV-1 in Resting CD4⁺ T Cells Using a Limiting Dilution Coculture Assay.

Author information

1
Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA.
2
Department of Biomedical Informatics, Columbia University Medical Center, New York, NY, 10032, USA.
3
Howard Hughes Medical Institute, Baltimore, MD, 21205, USA.
4
Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA. jsilicia@jhmi.edu.

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.

KEYWORDS:

HIV-1; Latency; Latent reservoir; Viral outgrowth assay

PMID:
26714716
DOI:
10.1007/978-1-4939-3046-3_16
[Indexed for MEDLINE]
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