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

GM Laird, DIS Rosenbloom, J Lai, RF Siliciano… - HIV protocols, 2016 - Springer
HIV protocols, 2016Springer
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 …
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.
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