HIV latency can be established in proliferating and nonproliferating resting CD4+ T cells in vitro
| Autor: | Damian F. J. Purcell, Sharon R Lewin, Samantha Adikari, Wan-Jung Cheng, Michael A Moso, Anne Ellett, Georges Khoury, Lachlan Robert Gray, Judy Chang, John Zaunders, Jenny L. Anderson, Melissa J Churchill, Jonathan C. Jacobson, Paul U. Cameron, Suha Saleh, Hao K. Lu |
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| Rok vydání: | 2019 |
| Předmět: |
Antigens
Differentiation T-Lymphocyte CD4-Positive T-Lymphocytes 0301 basic medicine Immunology Article Flow cytometry Green fluorescent protein 03 medical and health sciences 0302 clinical medicine Antigen Antigens CD Virus latency medicine Humans Immunology and Allergy Lectins C-Type Luciferase 030212 general & internal medicine IL-2 receptor Cells Cultured Cell Proliferation Staining and Labeling medicine.diagnostic_test Chemistry Interleukin-2 Receptor alpha Subunit HIV HLA-DR Antigens T lymphocyte Flow Cytometry medicine.disease Molecular biology In vitro Virus Latency 030104 developmental biology Infectious Diseases |
| Zdroj: | AIDS. 33:199-209 |
| ISSN: | 0269-9370 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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