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A novel dual luciferase assay for the simultaneous monitoring of HIV infection and cell viability.

Authors :
Mitsuki YY
Yamamoto T
Mizukoshi F
Momota M
Terahara K
Yoshimura K
Harada S
Tsunetsugu-Yokota Y
Source :
Journal of virological methods [J Virol Methods] 2016 May; Vol. 231, pp. 25-33. Date of Electronic Publication: 2016 Feb 17.
Publication Year :
2016

Abstract

Human immunodeficiency virus type 1 (HIV-1) reporter cell lines are critical tools for drug development. However, one disadvantage of HIV-1 reporter cell lines is that reductions in reporter gene activity need to be normalized to cytotoxicity, i.e., live cell numbers. Here, we developed a dual luciferase assay based on a R. reniformis luciferase (hRLuc)-expressing R5-type HIV-1 (NLAD8-hRLuc) and a CEM cell line expressing CCR5 and firefly luciferase (R5CEM-FiLuc). The NLAD8-hRLuc reporter virus was replication competent in peripheral blood mononuclear cells. The level of hRLuc was correlated with p24 antigen levels (p<0.001, R=0.862). The target cell line, R5CEM-FiLuc, stably expressed the firefly luciferase (FiLuc) reporter gene and allowed the simultaneous monitoring of compound cytotoxicity. The dual reporter assay combining a NLAD8-hRLuc virus with R5CEM-FiLuc cells permitted the accurate determination of drug susceptibility for entry, reverse transcriptase, integrase, and protease inhibitors at different multiplicities of infection. This dual reporter assay provides a rapid and direct method for the simultaneous monitoring of HIV infection and cell viability.<br /> (Copyright © 2016 Elsevier B.V. All rights reserved.)

Details

Language :
English
ISSN :
1879-0984
Volume :
231
Database :
MEDLINE
Journal :
Journal of virological methods
Publication Type :
Academic Journal
Accession number :
26898957
Full Text :
https://doi.org/10.1016/j.jviromet.2016.02.006