A novel HIV-1 antiviral high throughput screening approach for the discovery of HIV-1 inhibitors

• Published in: Antiviral research Vol. 65; no. 2; pp. 107 - 116
• Main Authors: Blair, Wade S., Isaacson, Jason, Li, Xinqiang, Cao, Joan, Peng, Qinghai, Kong, George F.Z., Patick, Amy K.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01.02.2005; Elsevier
• Subjects: Animals; Anti-HIV Agents - pharmacology; Antibiotics. Antiinfectious agents. Antiparasitic agents; Antiviral agents; Antiviral screen; Biological and medical sciences; Cell Line; Cell-based assay; Genes, Reporter; HeLa Cells; HIV; HIV-1 - drug effects; HIV-1 - genetics; Human immunodeficiency virus 1; Humans; Luciferases, Firefly - genetics; Medical sciences; Microbial Sensitivity Tests - methods; Microbial Sensitivity Tests - statistics & numerical data; Pharmacology. Drug treatments; Reporter virus; Reproducibility of Results; Antiviral screen; Cell-based assay; Reporter virus; HIV; High throughput screening; Immunopathology; Antiretroviral agent; HIV-1 virus; Retroviridae; AIDS; Immune deficiency; Lentivirus; In vitro; Infection; Virus; Viral disease; Antiviral; Human immunodeficiency virus
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/j.antiviral.2004.11.001

Antiviral high throughput screens remain a viable option for identifying novel target inhibitors. However, few antiviral screens have been reduced to practice on an industrial scale. In this study, we describe an HIV-1 dual reporter assay that allows for the simultaneous evaluation of the potential antiviral activities and cytotoxicities of compounds in a high throughput screen (HTS) format. We validate the assay with known HIV-1 inhibitors and show that the antiviral and cytotoxic activities of compounds are reproducibly measured under screening conditions. In addition, we show that the assay exhibits parameters (e.g., signal-to-background ratios and Z′ coefficients) suitable for high throughout screening. In a pilot screen, we demonstrate that non-specific or cytotoxic compounds represent a significant fraction of the hits identified in an antiviral screen and that these false positives are identified and deprioritized by the HIV-1 dual reporter assay at the primary screening step. We propose that the HIV-1 dual reporter assay represents a novel approach to HIV-1 antiviral screening that allows for the effective execution of industrial scale HTS campaigns with significantly greater returns on resource investment when compared to previous methods.