Single-cell characterization and quantification of translation-competent viral reservoirs in treated and untreated HIV infection.

Details

Serval ID
serval:BIB_E68F137DDCFC
Type
Article: article from journal or magazin.
Collection
Publications
Title
Single-cell characterization and quantification of translation-competent viral reservoirs in treated and untreated HIV infection.
Journal
PLoS pathogens
Author(s)
Pardons M., Baxter A.E., Massanella M., Pagliuzza A., Fromentin R., Dufour C., Leyre L., Routy J.P., Kaufmann D.E., Chomont N.
ISSN
1553-7374 (Electronic)
ISSN-L
1553-7366
Publication state
Published
Issued date
02/2019
Peer-reviewed
Oui
Volume
15
Number
2
Pages
e1007619
Language
english
Notes
Publication types: Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Abstract
The phenotypic characterization of the cells in which HIV persists during antiretroviral therapy (ART) remains technically challenging. We developed a simple flow cytometry-based assay to quantify and characterize infected cells producing HIV proteins during untreated and treated HIV infection. By combining two antibodies targeting the HIV capsid in a standard intracellular staining protocol, we demonstrate that p24-producing cells can be detected with high specificity and sensitivity in the blood from people living with HIV. In untreated individuals, the frequency of productively infected cells strongly correlated with plasma viral load. Infected cells preferentially displayed a transitional memory phenotype and were enriched in Th17, peripheral Tfh and regulatory T cells subsets. These cells also preferentially expressed activation markers (CD25, HLA-DR, Ki67), immune checkpoint molecules (PD-1, LAG-3, TIGIT, Tim-3) as well as the integrins α4β7 and α4β1. In virally suppressed individuals on ART, p24-producing cells were only detected upon stimulation (median frequency of 4.3 p24+ cells/106 cells). These measures correlated with other assays assessing the size of the persistent reservoir including total and integrated HIV DNA, Tat/rev Induced Limiting Dilution Assay (TILDA) and quantitative viral outgrowth assay (QVOA). In ART-suppressed individuals, p24-producing cells preferentially displayed a transitional and effector memory phenotype, and expressed immune checkpoint molecules (PD-1, TIGIT) as well as the integrin α4β1. Remarkably, α4β1 was expressed by more than 70% of infected cells both in untreated and ART-suppressed individuals. Altogether, these results highlight a broad diversity in the phenotypes of HIV-infected cells in treated and untreated infection and suggest that strategies targeting multiple and phenotypically distinct cellular reservoirs will be needed to exert a significant impact on the size of the reservoir.
Keywords
Adult, Anti-Retroviral Agents, CD4-Positive T-Lymphocytes, Disease Reservoirs/virology, Female, Flow Cytometry/methods, HIV/pathogenicity, HIV/physiology, HIV Core Protein p24, HIV Infections/immunology, HIV Infections/virology, HIV-1/immunology, HIV-1/pathogenicity, Humans, Integrin alpha4beta1/metabolism, Male, Middle Aged, Phenotype, RNA, Viral, Single-Cell Analysis/methods, T-Lymphocyte Subsets, Viral Load, Virus Latency
Pubmed
Web of science
Open Access
Yes
Create date
09/05/2023 12:59
Last modification date
29/11/2024 17:15
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