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AIDS. 2019 Feb 1;33(2):199-209. doi: 10.1097/QAD.0000000000002075.

HIV latency can be established in proliferating and nonproliferating resting CD4+ T cells in vitro: implications for latency reversal.

Author information

1
The Peter Doherty Institute for Infection and Immunity, University of Melbourne and Royal Melbourne Hospital.
2
Department of Infectious Diseases, Alfred Hospital and Monash University.
3
Centre for Biomedical Research, Burnet Institute.
4
Department of Microbiology and Immunology at The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Melbourne, Victoria.
5
Kirby Institute, University of New South Wales.
6
Centre for Applied Medical Research, St. Vincent's Hospital, Sydney, New South Wales.
7
School of Health and Biomedical Sciences, RMIT University, Bundoora, Victoria, Australia.

Abstract

OBJECTIVE:

To determine whether latency can be established and reversed in both proliferating and nonproliferating CD4 T cells in the same model in vitro.

METHODS:

Activated CD4 T cells were infected with either a nonreplication competent, luciferase reporter virus or wild-type full-length enhanced green fluorescent protein (EGFP) reporter virus and cultured for 12 days. The cells were then sorted by flow cytometry to obtain two distinct T-cell populations that did not express the T-cell activation markers, CD69, CD25 and human leukocyte antigen (HLA)-DR: CD69CD25HLA-DR small cells (nonblasts) that had not proliferated in vitro following mitogen stimulation and CD69CD25HLA-DR large cells (which we here call transitional blasts) that had proliferated. The cells were then reactivated with latency-reversing agents and either luciferase or EGFP quantified.

RESULTS:

Inducible luciferase expression, consistent with latent infection, was observed in nonblasts and transitional blasts following stimulation with either phorbol-myristate-acetate/phytohemagglutinin (3.8 ± 1 and 2.9 ± 0.5 fold above dimethyl sulfoxide, respectively) or romidepsin (2.1 ± 0.6 and 1.8 ± 0.2 fold above dimethyl sulfoxide, respectively). Constitutive expression of luciferase was higher in transitional blasts compared with nonblasts. Using wild-type full-length EGFP reporter virus, inducible virus was observed in nonblasts but not in transitional blasts. No significant difference was observed in the response to latency-reversing agents in either nonblasts or transitional blasts.

CONCLUSION:

HIV latency can be established in vitro in resting T cells that have not proliferated (nonblasts) and blasts that have proliferated (transitional blasts). This model could potentially be used to assess new strategies to eliminate latency.

PMID:
30562171
PMCID:
PMC6319264
[Available on 2020-02-01]
DOI:
10.1097/QAD.0000000000002075

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