Yeast cells as a tool for analysis of HIV-1 protease susceptibility to protease inhibitors, a comparative study.
HIV develops drug resistance at a high rate under drug selection pressure. Resistance tests are recommended to help physicians optimize antiretroviral drug therapies. For this purpose, genotypic and phenotypic tests have been developed. In order to propose a new phenotypic test that will be less laborious, expensive, and time consuming than the standard ones, a new procedure to measure HIV-1 protease susceptibility to protease inhibitor (PIs) in Saccharomyces cerevisiae yeast cells was developed. This procedure is based on HIV-1 protease expression in yeast. While the viral protein induces yeast cell death, its inhibition by PIs in the culture medium allows the cell to grow in a dose-dependent manner. In a comparative study of standard genotypic analysis vs. yeast cell-based phenotypic tests, performed on HIV-1 protease coding DNA in 17 different plasma samples from infected individuals, a clear match was found between the results obtained using the two technologies. This suggests that the yeast-based procedure is at least as accurate as standard genotypic test in defining susceptibility to protease inhibitors. This encouraging result should be the basis for large-scale validation of the new phenotypic resistance test.
Service des Maladies Infectieuses Hôpital de la Conception, 147 Bd Baille, 13385 Marseille Cedex 05, France. Electronic address: isabelle.ravaux@ap-hm.fr.
Isabelle
I
Ravaux
Christelle
C
Perrin-East
Coralie
C
Attias
Jacqueline
J
Cottalorda
Jacques
J
Durant
Pierre
P
Dellamonica
Pablo
P
Gluschankof
Andreas
A
Stein
Catherine
C
Tamalet
D019380:Anti-HIV Agents / Q000494:pharmacology*; D016333:HIV Protease / Q000096:biosynthesis* / Q000235:genetics / Q000633:toxicity; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D015497:HIV-1 / Q000201:enzymology*; D006801:Humans; D008826:Microbial Sensitivity Tests / Q000379:methods*; D050296:Microbial Viability / Q000187:drug effects*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000633:toxicity; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology
D019380:Anti-HIV Agents; D017320:HIV Protease Inhibitors; D011994:Recombinant Proteins; D016333:HIV Protease; C514803:p16 protease, Human immunodeficiency virus 1
ATV/r; DRV/r; FPV; Fosamprenavir; HIV-1 protease resistance; LPV/r; NFV; Nelfinavir; PI; Phenotypic test; Protease inhibitors; S. cerevisiae; SQV/r; Saquinavir boosted saquinavir; boosted Atazanavir; boosted Darunavir; boosted Lopinavir; protease inhibitor