A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells
Authors
Gludish, David W.
Jarvis, Isobel
Boliar, Saikat
Wills, Mark R.
Lever, Andrew M. L.
Mok, Hoi-Ping
Publication Date
2019-12-18Journal Title
Scientific Reports
Publisher
Nature Publishing Group UK
Volume
9
Issue
1
Language
en
Type
Article
This Version
VoR
Metadata
Show full item recordCitation
Salasc, F., Gludish, D. W., Jarvis, I., Boliar, S., Wills, M. R., Russell, D. G., Lever, A. M. L., & 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)https://doi.org/10.1038/s41598-019-55596-8
Description
Funder: DH | National Institute for Health Research (NIHR); doi: https://doi.org/10.13039/501100000272
Funder: U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (NIAID)
Abstract
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.
Keywords
Article, /631/1647/2234, /631/326/596/1787, /13/31, /14/63, /13/109, /38/1, article
Sponsorship
U.S. Department of Health & Human Services | NIH | NIH Office of the Director (OD) (T32OD011182)
RCUK | MRC | Medical Research Foundation (MR/S009752/1)
U.S. Department of Health & Human Services | NIH | National Institute of Allergy and Infectious Diseases (NIAID) (AI118582, AI136097)
RCUK | Medical Research Council (MRC) (MR/N02043X/1)
Identifiers
s41598-019-55596-8, 55596
External DOI: https://doi.org/10.1038/s41598-019-55596-8
This record's URL: https://www.repository.cam.ac.uk/handle/1810/315442
Rights
Licence:
http://creativecommons.org/licenses/by/4.0/