title: Development of a human immunodeficiency virus vector-based, single-cycle assay for evaluation of anti-integrase compounds.
abstract: Therapeutic strategies aimed at inhibiting human immunodeficiency virus type 1 (HIV-1) replication employ a combination of drugs targeted to two viral enzymes (reverse transcriptase and protease) and to the viral entry/fusion step. However, the high propensity of HIV-1 to develop resistance makes the development of novel compounds targeting different steps of the HIV-1 life cycle essential. Among these, integrase (IN) inhibitors have successfully passed the early phases of clinical development. By preventing integration, IN inhibitors preclude viral replication while allowing production of extrachromosomal forms of viral DNA (E-DNA). Here, we describe an improved and standardized assay aimed at evaluating IN inhibitors by taking advantage of the transcriptional activity of E-DNA produced by HIV-derived vectors in the absence of replication-competent virus. In this context, the use of the firefly luciferase gene as a reporter gene provides a rapid and quantitative measure of viral-vector infectivity, thus making it a safe and cost-effective assay for evaluating novel IN inhibitors.
authors.affiliation: National AIDS Center, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161, Rome, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Bona
authors.forename: Mauro
authors.initials: M
authors.lastname: Andreotti
authors.forename: Viviana
authors.initials: V
authors.lastname: Buffa
authors.forename: Pasqualina
authors.initials: P
authors.lastname: Leone
authors.forename: Clementina Maria
authors.initials: CM
authors.lastname: Galluzzo
authors.forename: Roberta
authors.initials: R
authors.lastname: Amici
authors.forename: Lucia
authors.initials: L
authors.lastname: Palmisano
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Mancini
authors.forename: Zuleika
authors.initials: Z
authors.lastname: Michelini
authors.forename: Roberto
authors.initials: R
authors.lastname: Di Santo
authors.forename: Roberta
authors.initials: R
authors.lastname: Costi
authors.forename: Alessandra
authors.initials: A
authors.lastname: Roux
authors.forename: Yves
authors.initials: Y
authors.lastname: Pommier
authors.forename: Christophe
authors.initials: C
authors.lastname: Marchand
authors.forename: Stefano
authors.initials: S
authors.lastname: Vella
authors.forename: Andrea
authors.initials: A
authors.lastname: Cara
mesh_terms: D002460:Cell Line; D005822:Genetic Vectors; D019427:HIV Integrase / Q000187:drug effects; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics; D006801:Humans; D020128:Inhibitory Concentration 50; D016086:Lentivirus / Q000235:genetics; D007963:Leukocytes, Mononuclear / Q000821:virology; D008156:Luciferases / Q000378:metabolism; D008826:Microbial Sensitivity Tests / Q000379:methods*; D014161:Transduction, Genetic
chemical_list: D019428:HIV Integrase Inhibitors; D008156:Luciferases; D019427:HIV Integrase