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A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells.

Accepted version
Peer-reviewed

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Authors

Gludish, David W 
Jarvis, Isobel 
Boliar, Saikat 
Wills, Mark R 

Abstract

Understanding the mechanisms involved in HIV infection and latency, and development of a cure, rely on the availability of sensitive research tools such as indicator cells, which allow rigorous quantification of viral activity. Here we describe the construction and validation of a novel dual-indicator cell line, Sup-GGR, which offers two different readouts to quantify viral replication. A construct expressing both Gaussia luciferase and hrGFP in a Tat- and Rev-dependent manner was engineered into SupT1-CCR5 to create Sup-GGR cells. This cell line supports the replication of both X4 and R5-tropic HIV as efficiently as its parental cell line, SupT1-CCR5, and allows repeated sampling without the need to terminate the culture. Sup-GGR demonstrates comparable sensitivity and similar kinetics in virus outgrowth assays (VOA) to SupT1-CCR5 using clinical samples. However the Gaussia luciferase reporter is significantly less labor-intensive and allows earlier detection of reactivated latent viruses compared to the conventional HIV p24 ELISA assay. The Sup-GGR cell line constitutes a versatile new tool for HIV research and clinical trials.

Description

Keywords

CD4-Positive T-Lymphocytes, Cell Line, Disease Reservoirs, HIV Infections, HIV-1, Humans, Luciferases, Virus Latency, Virus Replication

Journal Title

Sci Rep

Conference Name

Journal ISSN

2045-2322
2045-2322

Volume Title

9

Publisher

Springer Science and Business Media LLC

Rights

All rights reserved
Sponsorship
Medical Research Council (MR/S009752/1)
Medical Research Council (MR/N02043X/1)
Medical Research Council (MR/K021087/1)
National Institutes for Health Research, UK (NIHR, Cambridge Biomedical Research Centres), the Cambridge Clinical Academic Reserve and the Medical Research Council, UK (MR/N02043X/1) . This work was also supported by the National Institute of Health (AI118582 and AI36097).