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

  • Protocol
HIV Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1354))

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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Author information

Authors and Affiliations

  1. Department of Medicine, Johns Hopkins University School of Medicine, 733 N. Broadway, Baltimore, MD, 21205, USA

    Gregory M. Laird, Jun Lai, Robert F. Siliciano & Janet D. Siliciano

  2. Department of Biomedical Informatics, Columbia University Medical Center, New York, NY, 10032, USA

    Daniel I. S. Rosenbloom

  3. Howard Hughes Medical Institute, Baltimore, MD, 21205, USA

    Robert F. Siliciano

Authors
  1. Gregory M. Laird
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  2. Daniel I. S. Rosenbloom
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  3. Jun Lai
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  4. Robert F. Siliciano
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  5. Janet D. Siliciano
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Corresponding author

Correspondence to Janet D. Siliciano .

Editor information

Editors and Affiliations

  1. Department of Microbiology & Immunology, Albert Einstein College of Medicine, BRONX, New York, USA

    Vinayaka R. Prasad

  2. Department of Genetics, Albert Einstein College of Medicine, BRONX, New York, USA

    Ganjam V. Kalpana

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Laird, G.M., Rosenbloom, D.I.S., Lai, J., Siliciano, R.F., Siliciano, J.D. (2016). Measuring the Frequency of Latent HIV-1 in Resting CD4+ T Cells Using a Limiting Dilution Coculture Assay. In: Prasad, V., Kalpana, G. (eds) HIV Protocols. Methods in Molecular Biology, vol 1354. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3046-3_16

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  • DOI: https://doi.org/10.1007/978-1-4939-3046-3_16

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3045-6

  • Online ISBN: 978-1-4939-3046-3

  • eBook Packages: Springer Protocols

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