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Updated: Aug 6, 2026
High Throughput In Vitro Assessment of Latency Reversing Agents on HIV Transcription and Splicing
Published on: January 22, 2019
A novel in vitro system to generate and study latently HIV-infected long-lived normal CD4+ T-lymphocytes
Gautam K Sahu1, Kyeongeun Lee, Jiaxiang Ji
1Department of Microbiology and Immunology, The University of Texas Medical Branch, 301 University Blvd, BSB #3.132, Galveston, TX 77555, USA.
Insights
Researchers developed a novel in vitro model to study HIV latency. This system significantly increases the frequency of latently infected CD4+ T-cells, aiding research into HIV persistence and production.
Area of Science:
- Immunology
- Virology
- Cell Biology
Background:
- Studying HIV latency in resting CD4+ T-lymphocytes is challenging due to their low frequency in patients.
- Understanding HIV reactivation mechanisms requires accessible models of latently infected cells.
Purpose of the Study:
- To develop an in vitro system for generating a higher frequency of latently HIV-infected, long-lived quiescent CD4+ T-cells.
- To facilitate research into the mechanisms of HIV latency and persistence.
Main Methods:
- An in vitro culture system was established for long-term survival of post-activated CD4+ T-cells in a quiescent state.
- This system was used to generate latently infected quiescent CD4+ T-cells from HIV-infected activated T-lymphocytes.
Main Results:
- The in vitro system yielded a ~1000- to 10,000-fold increase in latently infected CD4+ T-cells compared to patient samples.
- 5-10% of these long-term surviving, non-cycling T-cells continuously produced infectious HIV at low levels.
- HIV production was observed without requiring full cellular activation.
Conclusions:
- The developed model system significantly enhances the study of latently HIV-infected quiescent CD4+ T-lymphocytes.
- This model aids in understanding persistent low-level HIV production from infected T-cells.
- It provides a valuable tool for investigating HIV latency and reactivation mechanisms.
Abstract:
Studies of mechanisms of HIV-latency and its reactivation in long-lived resting CD4+ T-lymphocytes in patients have been limited due to the very low frequency of these cells ( approximately 1-10 cells per 10(6) CD4+ T-cells). To circumvent this obstacle, an in vitro culture system for post-activation long-term survival of normal CD4+ T-cells in a quiescent (non-cycling) state was developed and used to generate latently infected, long-lived quiescent CD4+ T-cells from HIV-infected, activated normal CD4+ T-lymphocytes. This yielded a frequency of approximately 5x10(4) latently infected cells per 10(6) cells in culture, which is approximately 10(3)- to 10(4)-fold higher than that available from patients. Moreover, 5-10% of long-term surviving non-cycling T-cells were found to make infectious HIV continuously at low levels, showing that HIV production from infected T-cells does not require full cellular activation. This model system should facilitate studies of long-lived, latently infected and persistently HIV-producing quiescent normal CD4+ T-lymphocytes.
