A fluorescence-based high-throughput screening assay to identify HIV-1 inhibitors

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1030; p. 3
• Main Authors: Geluykens, Peggy, Van Acker, Koen, Vingerhoets, Johan, Van den Eynde, Christel, Van Loock, Marnix, Dams, Géry
• Format: Journal Article
• Language: English
• Published: United States 2013
• Subjects: Anti-HIV Agents - pharmacology; Gene Expression; Genes, Reporter; High-Throughput Screening Assays - methods; HIV-1 - drug effects; Humans; Microscopy, Fluorescence; Virus Replication - drug effects
• ISSN: 1940-6029
• DOI: 10.1007/978-1-62703-484-5_1

Highly active antiretroviral therapy (HAART) dramatically increases the long-term survival rates of human immunodeficiency virus type 1 (HIV-1) infected patients. Yet, poor adherence to therapy, adverse effects and the occurrence of resistant viruses can compromise the efficacy of HAART regiments. Therefore, there remains a clear unmet medical need for novel drugs and treatment options. In this chapter, we describe an HIV-1 antiviral high-throughput screening assay based on an HIV-1 permissive T lymphoblastoid MT4 cell line, stably transfected with a construct carrying an HIV-1 long terminal repeat promoter driving the expression of a reporter gene (enhanced green fluorescent protein). This assay runs in a 384-well format and enables the identification of HIV-1 inhibitors during a high-throughput screening campaign. In parallel, a cytotoxicity assay is performed to evaluate the compound-related in vitro toxicity.