title: High-throughput human immunodeficiency virus type 1 (HIV-1) full replication assay that includes HIV-1 Vif as an antiviral target.
abstract: Antiviral screens have proved useful for the identification of novel human immunodeficiency virus type 1 (HIV-1) inhibitors. In this study, we describe an HIV-1 full replication (HIV-1 Rep) assay that incorporates all of the targets required for replication in T-cell lines, including the HIV-1 Vif gene. The HIV-1 Rep assay was designed to exhibit optimal sensitivity to late-stage as well as early-stage inhibitors to maximize the likelihood of identification of novel target antiviral compounds in a screen. In addition, the flexibility of the HIV-1 Rep assay allows the rapid evaluation of antiviral compounds against different virus strains in different T-cell lines without significant modification of the assay format. We demonstrate that the HIV-1 Rep assay exhibits characteristics (e.g., a favorable Z' value) compatible with high-throughput screening in a 384-well format. The utility of the HIV-1 Rep assay was demonstrated in a high-throughput screen of >10(6) compounds. To our knowledge, this study represents the first example of an HIV-1 antiviral screen that includes Vif as a functional target and was executed on an industrial scale.
authors.affiliation: Department of Virology, Pfizer Global Research and Development, La Jolla Laboratories, 10777 Science Center Dr. (CB1), San Diego, CA 92121-1111, USA.
authors.forename: Joan
authors.initials: J
authors.lastname: Cao
authors.forename: Jason
authors.initials: J
authors.lastname: Isaacson
authors.forename: Amy K
authors.initials: AK
authors.lastname: Patick
authors.forename: Wade S
authors.initials: WS
authors.lastname: Blair
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D016346:Gene Products, vif / Q000235:genetics*; D016655:HIV Core Protein p24 / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D016013:Likelihood Functions; D010957:Plasmids / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects*; D054320:vif Gene Products, Human Immunodeficiency Virus
chemical_list: D000998:Antiviral Agents; D016346:Gene Products, vif; D016655:HIV Core Protein p24; D054320:vif Gene Products, Human Immunodeficiency Virus