1. A novel, sensitive dual-indicator cell line for detection and quantification of inducible, replication-competent latent HIV-1 from reservoir cells
- Author
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David W. Gludish, Saikat Boliar, Fanny Salasc, Hoi Ping Mok, Mark R. Wills, Isobel Jarvis, David G. Russell, Andrew M. L. Lever, Salasc, Fanny [0000-0002-6574-1358], Russell, David G [0000-0002-9748-750X], Apollo - University of Cambridge Repository, Russell, David G. [0000-0002-9748-750X], and Boliar, Saikat [0000-0002-3340-5368]
- Subjects
0301 basic medicine ,CD4-Positive T-Lymphocytes ,viruses ,030106 microbiology ,Human immunodeficiency virus (HIV) ,lcsh:Medicine ,HIV Infections ,13/109 ,medicine.disease_cause ,Virus Replication ,Virus ,Cell Line ,03 medical and health sciences ,Gaussia ,38/1 ,Replication (statistics) ,631/326/596/1787 ,medicine ,Humans ,Luciferase ,Latency (engineering) ,lcsh:Science ,Luciferases ,631/1647/2234 ,Disease Reservoirs ,Multidisciplinary ,Retrovirus ,biology ,lcsh:R ,article ,virus diseases ,biology.organism_classification ,Virology ,3. Good health ,Virus Latency ,13/31 ,030104 developmental biology ,Viral replication ,Cell culture ,14/63 ,HIV-1 ,lcsh:Q ,Microbiology techniques - 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.
- Published
- 2019