title: Efficient identification of human immunodeficiency virus type 1 mutants resistant to a protease inhibitor by using a random mutant library.
abstract: Emergence of drug-resistant mutant viruses during the course of antiretroviral therapy is a major hurdle that limits the success of chemotherapeutic treatment to suppress human immunodeficiency virus type 1 (HIV-1) replication and AIDS progression. Development of new drugs and careful patient management based on resistance genotyping data are important for enhancing therapeutic efficacy. However, identifying changes leading to drug resistance can take years of clinical studies, and conventional in vitro assays are limited in generating reliable drug resistance data. Here we present an efficient in vitro screening assay for selecting drug-resistant variants from a library of randomly mutated HIV-1 strains generated by transposon-directed base-exchange mutagenesis. As a test of principle, we screened a library of mutant HIV-1 strains containing random mutations in the protease gene by using a reporter T-cell line in the presence of the protease inhibitor (PI) nelfinavir (NFV). Analysis of replicating viruses from a single round of infection identified 50 amino acid substitutions at 35 HIV-1 protease residue positions. The selected mutant viruses showed specific resistance to NFV and included most of the known NFV resistance mutations. Therefore, the new assay is efficient for identifying changes leading to drug resistance. The data also provide insights into the molecular mechanisms underlying the development of drug resistance.
authors.affiliation: UCLA School of Medicine, Los Angeles, CA 90095, USA.
authors.forename: Sanggu
authors.initials: S
authors.lastname: Kim
authors.forename: Yun-Cheol
authors.initials: YC
authors.lastname: Kim
authors.forename: Hangfei
authors.initials: H
authors.lastname: Qi
authors.forename: Kunkai
authors.initials: K
authors.lastname: Su
authors.forename: Sherie L
authors.initials: SL
authors.lastname: Morrison
authors.forename: Samson A
authors.initials: SA
authors.lastname: Chow
mesh_terms: D002460:Cell Line; D024882:Drug Resistance, Viral / Q000235:genetics; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics*; D006801:Humans; D009154:Mutation; D019888:Nelfinavir / Q000494:pharmacology
chemical_list: D017320:HIV Protease Inhibitors; D019888:Nelfinavir