TY - JOUR
T1 - Rapid phenotypic drug susceptibility assay for HIV-1 with a CCR5 expressing indicator cell line
AU - Pirounaki, Maria
AU - Vander Heyden, Nancy A.
AU - Arens, Max
AU - Ratner, Lee
N1 - Funding Information:
We would like to thank Anahid Birdwell for technical support, and Chia Hung, Suzanne Pontow, and Frosso Voulgaropoulou for critical reading of the manuscript. We acknowledge the support of Burroughs Wellcome, Abbott, and Merck for providing the drugs for this study and the Washington University Aids Clinical Trials Group for providing patient samples. MP was a fellow of the Infectious Diseases Division, Washington University School of Medicine, and was also supported by a scholarship from the Greek State Scholarship Foundation. The study was supported by grants from the Public Health Service.
PY - 2000/3
Y1 - 2000/3
N2 - Phenotypic drug susceptibility assays of human immunodeficiency virus type 1 (HIV-1) isolates generally use time-consuming, expensive assays with peripheral 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 gene β-galactosidase driven by the HIV-1 LTR, and quantitates infection within 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 (zidovudine, ZDV) and a non-nucleoside reverse transcriptase inhibitor (nevirapine, NVP) were performed with reference and primary isolates. The assay was modified into two steps for protease inhibitor (indivinavir, IDV and ritonavir, RTV) susceptibility assays. Primary isolates derived from drug naive patients displayed mean baseline 50% effective concentrations (EC50) of 0.14 μM for ZDV, 0.33 μM for NVP, and 0.02 μM for IDV. Isolates derived from patients under treatment displayed increased EC50 concentrations. The MAGI-CCR5 cell line offers a rapid, efficient, and reproducible method of testing a wide range of HIV-1 isolates for drug susceptibility. Copyright (C) 2000 Elsevier Science B.V.
AB - Phenotypic drug susceptibility assays of human immunodeficiency virus type 1 (HIV-1) isolates generally use time-consuming, expensive assays with peripheral 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 gene β-galactosidase driven by the HIV-1 LTR, and quantitates infection within 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 (zidovudine, ZDV) and a non-nucleoside reverse transcriptase inhibitor (nevirapine, NVP) were performed with reference and primary isolates. The assay was modified into two steps for protease inhibitor (indivinavir, IDV and ritonavir, RTV) susceptibility assays. Primary isolates derived from drug naive patients displayed mean baseline 50% effective concentrations (EC50) of 0.14 μM for ZDV, 0.33 μM for NVP, and 0.02 μM for IDV. Isolates derived from patients under treatment displayed increased EC50 concentrations. The MAGI-CCR5 cell line offers a rapid, efficient, and reproducible method of testing a wide range of HIV-1 isolates for drug susceptibility. Copyright (C) 2000 Elsevier Science B.V.
KW - HIV-1
KW - Indinavir
KW - Nevirapine
KW - Zidovudine
UR - http://www.scopus.com/inward/record.url?scp=0033977791&partnerID=8YFLogxK
U2 - 10.1016/S0166-0934(99)00163-9
DO - 10.1016/S0166-0934(99)00163-9
M3 - Article
C2 - 10716348
AN - SCOPUS:0033977791
SN - 0166-0934
VL - 85
SP - 151
EP - 161
JO - Journal of Virological Methods
JF - Journal of Virological Methods
IS - 1-2
ER -