Protocol

HIV Protocols

Volume 1354 of the series Methods in Molecular Biology pp 239-253

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

  • Gregory M. LairdAffiliated withDepartment of Medicine, Johns Hopkins University School of Medicine
  • , Daniel I. S. RosenbloomAffiliated withDepartment of Biomedical Informatics, Columbia University Medical Center
  • , Jun LaiAffiliated withDepartment of Medicine, Johns Hopkins University School of Medicine
  • , Robert F. SilicianoAffiliated withDepartment of Medicine, Johns Hopkins University School of MedicineHoward Hughes Medical Institute
  • , Janet D. SilicianoAffiliated withDepartment of Medicine, Johns Hopkins University School of Medicine Email author 

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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.

Key words

HIV-1 Latency Latent reservoir Viral outgrowth assay