Measuring the frequency of latent HIV-1 in resting CD4+ T cells using a limiting dilution coculture assay

Gregory M. Laird, Daniel I.S. Rosenbloom, Jun Lai, Robert F. Siliciano, Janet D. Siliciano

Research output: Chapter in Book/Report/Conference proceedingChapter

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 n 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.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages239-253
Number of pages15
DOIs
StatePublished - 2016

Publication series

NameMethods in Molecular Biology
Volume1354
ISSN (Print)1064-3745

Keywords

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

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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