Infection with a newly-designed dual fluorescent reporter HIV-1 effectively identifies latently infected CD4+ T cells

Abstract

The major barrier to curing HIV-1 infection is a small pool of latently infected cells that harbor replication-competent viruses, which are widely considered the origin of viral rebound when ART is interrupted. The difficulty of distinguishing latently infected cells from the vast majority of uninfected cells has represented a significant bottleneck precluding comprehensive understandings of HIV-1 latency. Here we reported and validated a newly-designed dual fluorescent reporter virus, DFV-B, infection with which in primary CD4+ T cells can directly label latently infected cells and generate a latency model that was highly physiological relevant. Applying DFV-B infection in Jurkat T cells, we generated a stable cell line model of HIV-1 latency with diverse viral integration sites. High-throughput compound screening with this model identified ACY-1215 as a potent latency reversing agent, which could be verified in other cell models and in primary CD4+ T cells from ART-suppressed individuals ex vivo. In summary, we have generated a meaningful and feasible model to directly study latently infected cells, which could open up new avenues to explore the critical events of HIV-1 latency and become a valuable tool for the research of AIDS functional cure.

Data availability

All data generated or analysed during this study are included in the manuscript.

Article and author information

Author details

  1. Jinfeng Cai

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  2. Hongbo Gao

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  3. Jiacong Zhao

    Institute of Human Virology, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  4. Shujing Hu

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  5. Xinyu Liang

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  6. Yanyan Yang

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  7. Zhunaglin Dai

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Zhongsi Hong

    Fifth Affiliated Hospital, Sun Yat-sen University, Zhuhai, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Kai Deng

    Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, China
    For correspondence
    dengkai6@mail.sysu.edu.cn
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9973-8130

Funding

National Natural Science Foundation of China (81672024)

  • Kai Deng

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Human subjects: Human subjects: The use of PBMCs from healthy individuals was approved by the Institutional Review Board of Guangzhou Blood Center (Guangzhou, Guangdong, China). We did not have any interaction with the healthy individuals or protected information, and therefore no informed consent was required. Chronically HIV-1-infected participants sampled by this study were recruited from The Fifth Affiliated Hospital of Sun Yat-sen University (Zhuhai, Guangdong, China). This study was approved by the Ethics Review Boards of the Fifth Affiliated Hospital of Sun Yat-sen University (2018K41-1). All the participants were given written informed consent with approval of the Ethics Committees. The enrollment of HIV-1-infected individuals was based on the criteria of prolonged suppression of plasma HIV-1 viremia on cART, which is undetectable plasma HIV-1 RNA levels (less than 50 copies/ml) for a minimum of 12 months, and having high CD4+ T cell count (at least 350 cells/mm3). All HIV-1-infected participants provided written informed consent for their participation in the study and agreed with the publication of the scientific results.

Reviewing Editor

  1. Jori Symons

Publication history

  1. Received: October 7, 2020
  2. Accepted: April 4, 2021
  3. Accepted Manuscript published: April 9, 2021 (version 1)
  4. Version of Record published: April 12, 2021 (version 2)

Copyright

© 2021, Cai et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Jinfeng Cai
  2. Hongbo Gao
  3. Jiacong Zhao
  4. Shujing Hu
  5. Xinyu Liang
  6. Yanyan Yang
  7. Zhunaglin Dai
  8. Zhongsi Hong
  9. Kai Deng
(2021)
Infection with a newly-designed dual fluorescent reporter HIV-1 effectively identifies latently infected CD4+ T cells
eLife 10:e63810.
https://doi.org/10.7554/eLife.63810

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