Cabana Sumsi M, Parera M, Clotet B, Martinez MA; International Conference on AIDS.

Int Conf AIDS. 2000 Jul 9-14; 13: abstract no. TuOrA346.

M. Cabana Sumsi, Fundacic Irsicaixa, Hosp. Universitare Germans Trias i Pujol, Ctra del Canyet S/N, 08916 Badalona, Spain, Tel.: +34 934 656 374, Fax: +34 934 653 968, E-mail: mcabana@ns.hugtip.scs.es

Human immunodeficiency virus type 1 (HIV-1) resistance to antiretroviral drugs is the main cause of patient treatment failure. Despite the problems associated with interpretation of HIV-1 resistance testing, resistance monitoring should help the rational design of initial or rescue antiretroviral therapies. We have developed a simple bacteriophage lambda-based genetic screen system for analysis of the activity and phenotype of the HIV-1 protease. This genetic screen system is based on the bacteriophage lambda regulatory circuit where the viral repressor cI is specifically cleaved to initiate lysogenic to lytic switch. Single critical mutations associated with resistance to protease inhibitors were introduced by site-directed mutagenesis. Lambda phages encoding HIV-1 proteases from laboratory generated mutants and from clinical samples were tested for their activity and phenotype. As expected, differences in proteolytic processing activity were seen among this group of HIV-1 proteases. Distinct susceptibilities to different drugs (saquinavir, ritonavir, indinavir and nelfinavir) were also detected among phages encoding HIV-1 proteases carrying different resistance mutations. Laboratory generated proteases encoding single mutations, didn't show as high resistance as did those from patients, which have accumulated various critical resistance mutations and secondary substitutions. In summary, we present here a simple, safe and rapid genetic screen system that may be used in predicting the activity and phenotype of HIV-1 proteases in the course of viral infection and antiretroviral therapy. This assay responds appropriately to well know HIV-1 protease inhibitors and can be used to search new protease inhibitors as well as novel drug-resistant mutants.

Publication Types:

• Meeting Abstracts

Keywords:

• Bacteriophage lambda
• Endopeptidases
• HIV Protease
• HIV-1
• Humans
• Indinavir
• Motor Activity
• Mutagenesis, Site-Directed
• Mutation
• Nelfinavir
• Phenotype
• Protease Inhibitors
• Ritonavir
• Saquinavir
• genetics

Other ID:

• GWAIDS0001304

UI: 102238795

From Meeting Abstracts