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Modeling HIV-1 Latency in Primary T Cells Using a Replication-Competent Virus
Laura J Martins1, Pawel Bonczkowski2, Adam M Spivak3
11 Division of Microbiology and Immunology, Department of Pathology, University of Utah School of Medicine , Salt Lake City, Utah.
Insights
This study presents a novel in vitro model for HIV-1 latency using replication-competent viruses and central memory CD4(+) T cells. The model effectively generates and reactivates latent HIV-1 infections, mimicking in vivo conditions with antiretroviral therapy.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Studying HIV-1 latency in vivo is challenging due to the extremely low frequency of latently infected cells.
- In vitro models are crucial, but often use defective viruses, limiting their recapitulation of natural infection dynamics.
- Replication-competent viruses offer a more accurate model but necessitate the use of antiretrovirals in culture.
Purpose of the Study:
- To develop and validate an in vitro model for studying HIV-1 latency using replication-competent viruses.
- To generate latently infected cells that can be reactivated under conditions mimicking antiretroviral treatment (ART).
- To implement methods for efficient removal of productively infected cells and increased throughput in latency studies.
Main Methods:
- Utilized cultured central memory CD4(+) T cells and replication-competent HIV-1 for infection.
- Incorporated antiretroviral drugs during latency establishment and reactivation phases.
- Developed a method to eliminate productively infected cells by exploiting HIV-1-induced CD4 downregulation.
- Employed a GFP-encoding virus to facilitate high-throughput analysis.
Main Results:
- Successfully generated latently infected CD4(+) T cells using replication-competent HIV-1.
- Demonstrated successful reactivation of latent HIV-1 in the presence of antiretroviral drugs.
- Validated a method for removing productively infected cells, enhancing the study of latent reservoirs.
- Showcased the utility of GFP-encoding viruses for increased experimental throughput.
Conclusions:
- The developed model accurately recapitulates key aspects of HIV-1 latency and reactivation in vitro.
- This model provides a valuable tool for investigating latency-reversing strategies and the impact of ART on viral reservoirs.
- The methodology facilitates more efficient and relevant research into potential HIV-1 cures.
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.
