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

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


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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / virology*
  • Cells, Cultured
  • Down-Regulation
  • Flow Cytometry
  • Green Fluorescent Proteins / genetics
  • HIV Infections / virology
  • HIV-1 / physiology*
  • Humans
  • Leukocytes, Mononuclear / virology*
  • Models, Biological*
  • Virus Activation / physiology*
  • Virus Latency / physiology*
  • Virus Replication / physiology


  • Green Fluorescent Proteins