A highly reliable, sensitive, flow cytometric/fluorometric assay for the evaluation of the anti-HIV activity of antiviral compounds in MT-4 cells.
Infection of human T4 lymphocyte MT-4 cells with human immunodeficiency virus (HIV) results in cell death 4-5 days after infection. We have now developed a highly sensitive and rapid procedure for estimating the cytopathic effect of HIV in MT-4 cells. This method is based on fluorometric as well as flow cytometric evaluation of HIV-infected MT-4 cultures. By the use of fluorescein diacetate (FDA), a non-fluorescent diacetyl fluorescein ester that becomes fluorescent upon hydrolysis by esterases present in the cytoplasm of viable cells, as few as 100-200 viable MT-4 cells can be accurately determined. Applying this new method to HIV-infected MT-4 cell cultures treated with differing concentrations of the potent anti-HIV agent azidothymidine (AZT), we obtained a virus-inhibitory dose-response comparable to those obtained by the conventional (labour-intensive and time-consuming) methods. The FDA-based cell viability assay appears particularly suited for the rapid, reliable and sensitive evaluation of potential anti-AIDS agents in cell culture.
D000163:Acquired Immunodeficiency Syndrome / Q000382:microbiology / Q000473:pathology; D015496:CD4-Positive T-Lymphocytes / Q000187:drug effects* / Q000382:microbiology; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003588:Cytopathogenic Effect, Viral / Q000187:drug effects; D005404:Fixatives; D005434:Flow Cytometry / Q000379:methods; D005452:Fluoresceins; D006678:HIV / Q000187:drug effects*; D006801:Humans; D007958:Leukocyte Count; D015215:Zidovudine / Q000494:pharmacology*
D005404:Fixatives; D005452:Fluoresceins; D015215:Zidovudine; C018506:diacetylfluorescein