A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells.
Nature Publishing Group
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Salasc, F., Gludish, D. W., Jarvis, I., Boliar, S., Wills, M., Russell, D. G., Lever, A., & et al. (2019). A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells.. Scientific reports, 9 (1), 19325. https://doi.org/10.1038/s41598-019-55596-8
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 qantify 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.
CD4-Positive T-Lymphocytes, Cell Line, Humans, HIV-1, HIV Infections, Luciferases, Disease Reservoirs, Virus Latency, Virus Replication
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).
MEDICAL RESEARCH COUNCIL (MR/N02043X/1)
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External DOI: https://doi.org/10.1038/s41598-019-55596-8
This record's URL: https://www.repository.cam.ac.uk/handle/1810/299145
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