Department of Public Health Sciences

Standardized Peripheral-blood Mononuclear Cell-culture Assay For Determination of Drug Susceptibilities of Clinical Human- Immunodeficiency-virus Type-1 Isolates

Japour, AJ, DL Mayers, VA Johnson, DR Kuritzkes, LA Beckett, JM Arduino, J Lane, RJ Black, PS Reichelderfer, RT Daquila, CS Crumpacker, H Balfour, A Erice, R Coombs, D Katzenstein, J Lathey, D Richman, K Mcintosh, S Rangan, R Reichman, W Scott, M Ussery, L Abrams, F McCutchan, D Burke, L Gardner, C Roberts, R Chung, C Hicks, E Shellie, A Fowler, L Merritt, M Fujimurajustice, N Ruiz, K Wagner and M Gail

Antimicrob. Agents Chemother.. 1993. 37(5):1095-1101.

A standardized antiviral drug susceptibility assay for clinical human immunodeficiency virus type 1 (HIV-1) isolates has been developed for use in clinical trials. The protocol is a two- step procedure that first involves cocultivation of patient infected peripheral blood mononuclear cells (PBMC) with seronegative phytohemag-glutinin-stimulated donor PBMC to obtain an HIV-1 stock. The virus stock is titrated for viral infectivity (50% tissue culture infective dose) by use of serial fourfold virus dilutions in donor PBMC. A standardized inoculum of 1,000 50% tissue culture infective doses per 10(6) cells is used in the second step of the procedure to acutely infect seronegative donor PBMC in a 7-day microtiter plate assay with triplicate wells containing zidovudine (ZDV) concentrations ranging from 0 to 5.0 muM. The ZDV 50% inhibitory concentrations (IC50) for reference ZDV-susceptible and ZDV-resistant HIV-1 isolates ranged from 0.002 to 0.113 muM and from 0.15 to >5.0 muM, respectively. Use of this consensus protocol reduced interlaboratory variability for ZDV IC50 determinations with reference HIV-1 isolates. Among eight laboratories, the coefficient of variation ranged from 0.85 to 1.25 with different PBMC protocols and was reduced to 0.39 to 0.98 with the standardized assay. Among the clinical HIV-1 isolates assayed by the standardized drug susceptibility assay, the median ZDV IC50 increased gradually with more ZDV therapy. This protocol provides an efficient and reproducible means to assess the in vitro susceptibility to antiretroviral agents of virtually all clinical HIV-1 isolates.

Keywords: Aging; Alcohol Drinking; Alzheimers Disease; Cognition; Prospective Studies; Longitudinal Studies; Risk Factors; Smoking, Alzheimers-disease; Men; Drinking, Hiv-1 Reverse-transcriptase; Zidovudine Azt; Reduced Sensitivity; Resistant; Inhibitors; Therapy; Invitro



Close Window

UC Davis Health System is pleased to provide this information for general reference purposes only. It should not be considered as a substitute for professional medical advice. You are urged to consult with your health care provider for diagnosis of and treatment for any health-related condition. The information provided herein may not and should not be used for diagnosis and treatment.

Reproduction of material on this web site is hereby granted solely for personal use. No other use of this material is authorized without prior written approval of UC Regents.