title: Evaluation of HIV-1 integrase inhibitors on human primary macrophages using a luciferase-based single-cycle phenotypic assay.
abstract: Macrophages represent an important site for productive infection of HIV-1 and the evaluation of integrase (IN) inhibitors on this cell subset is of fundamental importance. In this report, preclinical evaluation of IN inhibitors on primary human macrophages was attempted successfully using a 96-well microtiter phenotypic assay developed recently for the evaluation of IN inhibitors in a cell-based system by taking advantage of HIV-derived lentiviral vectors expressing luciferase. IN inhibitors were also tested using a lentiviral vector containing an IN with introduced T66I/S153Y mutations, known to affect the activity of azido-group-containing diketo acid (DKA) IN inhibitors. Utilizing different classes of HIV integrase inhibitors against the wild-type IN and the mutant mentioned above, some of the IN inhibitors used were also active on this particular mutant, suggesting that should HIV-1 develop additional or different mutations to become resistant to such anti-IN drugs, new drugs can be developed with a better resistance profile. This assay provides a standardized method for the preclinical evaluation of the efficacy of IN inhibitors on wild-type and mutated IN that can be adapted easily for the evaluation of anti-IN activity on IN sequences derived from patients.
authors.affiliation: Department of Therapeutic Research and Medicines Evaluation, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: Zuleika
authors.initials: Z
authors.lastname: Michelini
authors.forename: Clementina Maria
authors.initials: CM
authors.lastname: Galluzzo
authors.forename: Donatella R M
authors.initials: DR
authors.lastname: Negri
authors.forename: Pasqualina
authors.initials: P
authors.lastname: Leone
authors.forename: Roberta
authors.initials: R
authors.lastname: Amici
authors.forename: Roberta
authors.initials: R
authors.lastname: Bona
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Summa
authors.forename: Roberto
authors.initials: R
authors.lastname: Di Santo
authors.forename: Roberta
authors.initials: R
authors.lastname: Costi
authors.forename: Yves
authors.initials: Y
authors.lastname: Pommier
authors.forename: Christophe
authors.initials: C
authors.lastname: Marchand
authors.forename: Lucia
authors.initials: L
authors.lastname: Palmisano
authors.forename: Stefano
authors.initials: S
authors.lastname: Vella
authors.forename: Andrea
authors.initials: A
authors.lastname: Cara
mesh_terms: D019380:Anti-HIV Agents / Q000494:pharmacology*; D001681:Biological Assay / Q000379:methods; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D017930:Genes, Reporter; D005822:Genetic Vectors; D019427:HIV Integrase / Q000235:genetics / Q000378:metabolism*; D019428:HIV Integrase Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000254:growth & development*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D019380:Anti-HIV Agents; D019428:HIV Integrase Inhibitors; D050505:Mutant Proteins; D008156:Luciferases; D019427:HIV Integrase; C514801:p31 integrase protein, Human immunodeficiency virus 1