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Development of an HIV reporter virus that identifies latently infected CD4+ T cells

Eun Hye Kim, Lara Manganaro, Michael Schotsaert, Brian D. Brown, Lubbertus C.F. Mulder, Viviana Simon
doi: https://doi.org/10.1101/2022.01.17.476679
Eun Hye Kim
1Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, USA, 10029
3The Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, USA
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Lara Manganaro
4INGM-Istituto Nazionale Genetica Molecolare, Milan, Italy, 20122
5Department of Pharmacological and Biomolecular Sciences (DiSFeB), University of Milan, Milan, Italy
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Michael Schotsaert
1Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, USA, 10029
3The Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, USA
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Brian D. Brown
7Precision Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, USA, 10029
8Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, New York, 10029
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Lubbertus C.F. Mulder
1Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, USA, 10029
3The Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, USA
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Viviana Simon
1Department of Microbiology, Icahn School of Medicine at Mount Sinai, New York, USA, 10029
2Division of Infectious Disease, Department of Medicine, Icahn School of Medicine at Mount Sinai, New York, USA
3The Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, New York, USA
6Department of Pathology, Molecular and Cellular Medicine at Icahn School of Medicine at Mount Sinai, New York, USA, 10029
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  • For correspondence: viviana.simon@mssm.edu
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SUMMARY

There is no cure for HIV infection as the virus establishes a latent reservoir, which escapes highly active antiretroviral treatments. One major obstacle is the difficulty to identify cells that harbor latent proviruses. We devised a novel viral vector that carries a series of versatile reporter molecules that are expressed in an LTR-dependent or LTR-independent manner and allows to accurately distinguish productive from latent infection. Using primary human CD4+ T cells, we show that transcriptionally silent proviruses make up over 50% of all infected cells. These latently infected cells harbor proviruses, but lack evidence for viral transcription. LTR silent integrations occurred to variable degrees in all CD4+ T-subsets examined with CD4+ TEM and CD4+ Treg displaying the highest frequency. Viral vectors such as the one described here, permit interrogation HIV latency at a single-cell resolution, revealing mechanisms of latency establishment and allowing for the characterization of effective latency reversing agents.

Competing Interest Statement

The authors have declared no competing interest.

Copyright 
The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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Posted January 18, 2022.
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Development of an HIV reporter virus that identifies latently infected CD4+ T cells
Eun Hye Kim, Lara Manganaro, Michael Schotsaert, Brian D. Brown, Lubbertus C.F. Mulder, Viviana Simon
bioRxiv 2022.01.17.476679; doi: https://doi.org/10.1101/2022.01.17.476679
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Development of an HIV reporter virus that identifies latently infected CD4+ T cells
Eun Hye Kim, Lara Manganaro, Michael Schotsaert, Brian D. Brown, Lubbertus C.F. Mulder, Viviana Simon
bioRxiv 2022.01.17.476679; doi: https://doi.org/10.1101/2022.01.17.476679

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