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Modeling HIV-1 Latency in Primary T Cells Using a Replication-Competent Virus.
- Source :
-
AIDS research and human retroviruses [AIDS Res Hum Retroviruses] 2016 Feb; Vol. 32 (2), pp. 187-93. Date of Electronic Publication: 2015 Jul 14. - Publication Year :
- 2016
-
Abstract
- 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.
- Subjects :
- CD4-Positive T-Lymphocytes immunology
Cells, Cultured
Down-Regulation
Flow Cytometry
Green Fluorescent Proteins genetics
HIV Infections virology
Humans
Virus Replication physiology
CD4-Positive T-Lymphocytes virology
HIV-1 physiology
Leukocytes, Mononuclear virology
Models, Biological
Virus Activation physiology
Virus Latency physiology
Subjects
Details
- Language :
- English
- ISSN :
- 1931-8405
- Volume :
- 32
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- AIDS research and human retroviruses
- Publication Type :
- Academic Journal
- Accession number :
- 26171776
- Full Text :
- https://doi.org/10.1089/aid.2015.0106