New soluble-formazan assay for HIV-1 cytopathic effects: application to high-flux screening of synthetic and natural products for AIDS-antiviral activity.
We have developed an effective and optimally safe microculture method for rapid and convenient assay of the in vitro cytopathic effects of human immunodeficiency virus (HIV-1) on human lymphoblastoid or other suitable host cells. The assay procedure is applicable to the evaluation of drug effects on in vitro infections induced directly in cultured host cells by cell-free HIV-1 or by coculture with H9 cells chronically infected with HIV-1. The assay uses a newly developed tetrazolium reagent that is metabolically reduced by viable cells to yield a soluble, colored formazan product measurable by conventional colorimetric techniques. This simple microassay minimizes the number of plate manipulations typically required with other assay methods and, coupled with computerized data collection and analysis, facilitates large-scale screening of agents for potential antiviral activity. To support and enhance the discovery of new anti-HIV-1 agents, the National Cancer Institute is offering investigators worldwide the opportunity to submit new candidate agents for anti-HIV-1 screening with this method.
Weislow
O S
OS
Program Resources, Inc., National Cancer Institute-Frederick Cancer Research Facility, MD.
Kiser
R
R
Fine
D L
DL
Bader
J
J
Shoemaker
R H
RH
Boyd
M R
MR
D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D005562:Formazans / Q000378:metabolism; D015488:HIV Antigens / Q000096:biosynthesis; D016655:HIV Core Protein p24; D015497:HIV-1 / Q000187:drug effects*; D006801:Humans; D007202:Indicators and Reagents; D012191:Retroviridae Proteins / Q000096:biosynthesis; D013778:Tetrazolium Salts
D000998:Antiviral Agents; D005562:Formazans; D015488:HIV Antigens; D016655:HIV Core Protein p24; D007202:Indicators and Reagents; D012191:Retroviridae Proteins; D013778:Tetrazolium Salts; C059087:2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; C059088:1,5-bis(2-methoxy-4-nitro-5-sulfophenyl)-3-((phenylamino)carbonyl)formazan