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AIDS Res Hum Retroviruses. 2016 Feb;32(2):187-93. doi: 10.1089/aid.2015.0106. Epub 2015 Jul 14.

Modeling HIV-1 Latency in Primary T Cells Using a Replication-Competent Virus.

Author information

1 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
2 Department of Internal Medicine, HIV Translational Research Unit, Ghent University , Ghent, Belgium .
3 Division of Infectious Diseases, Department of Medicine, University of Utah School of Medicine , Salt Lake City, Utah.


HIV-1 latently infected cells in vivo can be found in extremely low frequencies. Therefore, in vitro cell culture models have been used extensively for the study of HIV-1 latency. Often, these in vitro systems utilize defective viruses. Defective viruses allow for synchronized infections and circumvent the use of antiretrovirals. In addition, replication-defective viruses cause minimal cytopathicity because they fail to spread and usually do not encode env or accessory genes. On the other hand, replication-competent viruses encode all or most viral genes and better recapitulate the nuances of the viral replication cycle. The study of latency with replication-competent viruses requires the use of antiretroviral drugs in culture, and this mirrors the use of antiretroviral treatment (ART) in vivo. We describe a model that utilizes cultured central memory CD4(+) T cells and replication-competent HIV-1. This method generates latently infected cells that can be reactivated using latency reversing agents in the presence of antiretroviral drugs. We also describe a method for the removal of productively infected cells prior to viral reactivation, which takes advantage of the downregulation of CD4 by HIV-1, and the use of a GFP-encoding virus for increased throughput.

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