Rapid phenotypic drug susceptibility assay for HIV-1 with a CCR5 expressing indicator cell line

Phenotypic drug susceptibility assays of human immunodeficiency virus type 1 (HIV-1) isolates generally use time-consuming, expensive assays with peri pheral blood mononuclear cells. A new HIV-1 indicator cell line, MAGI-CCR5, has been developed and applied for this purpose. This cell line expresses human CD4, the two major HIV-1 coreceptors, CCR5 and CXCR4, the reporter ge ne beta-galactosidase driven by the HIV-1 LTR, and quantitates infection wi thin 48 h. A panel of reference strains and primary HIV-1 isolates were all found to infect this cell line. Susceptibility assays with a nucleoside (z idovudine, ZDV) and a non-nucleoside reverse transcriptase inhibitor (nevir apine, NVP) were performed with reference and primary isolates. The assay w as modified into two steps for protease inhibitor (indivinavir, IDV and rit onavir, RTV) susceptibility assays. Primary isolates derived from drug naiv e patients displayed mean baseline 50% effective concentrations (EC50) of 0 .14 mu M for ZDV, 0.33 mu M for NVP, and 0.02 mu M for IDV. Isolates derive d from patients under treatment displayed increased EC50 concentrations. Th e MAGI-CCR5 cell line offers a rapid, efficient, and reproducible method of testing a wide range of HIV-1 isolates for drug susceptibility. (C) 2000 P ublished by Elsevier Science B.V. All rights reserved.