A simple and rapid method for preparation of HIV-expressing targets for functional assays.
Standardized samples of target cells expressing viral antigen are necessary to perform reproducible and comparable functional assays for immune responses to human immunodeficiency virus (HIV-1). HIV-infected cultured cell lines show wide variability in the number of HIV-expressing cells and in the amount of viral antigen per cell. They, as well as cells coated with the infectious virus, are a biohazard. 0.015% v/v beta-propiolactone (BPL) in buffered solution at pH 7.5 inactivates the infectivity of HIV but retains its antigenicity. BPL-inactivated HIV easily binds to T4 receptor bearing T-lymphocytes in a 30 min incubation at 22 degrees C. The process of adsorption is dose-dependent and results in a standardized and reproducible sample of target cells. Applications for the method are demonstrated and include: (1) preparation of target cells for assay of antibody-dependent cellular cytotoxicity (ADCC), and (2) characterization of antisera to HIV for their capacity to block HIV binding to cells.
Nishanian
P G
PG
Department of Microbiology and Immunology, UCLA School of Medicine 90024-1747.
Ojo-Amaize
E
E
Uittenbogaart
C H
CH
Giorgi
J V
JV
D000914:Antibodies, Viral / Q000032:analysis*; D000920:Antibody-Dependent Cell Cytotoxicity; D000956:Antigens, Viral / Q000276:immunology*; D002460:Cell Line; D046508:Culture Techniques / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D006678:HIV / Q000187:drug effects / Q000254:growth & development / Q000276:immunology*; D015483:HIV Antibodies; D006801:Humans; D008214:Lymphocytes / Q000276:immunology; D011420:Propiolactone / Q000494:pharmacology; D011991:Receptors, Virus / Q000032:analysis
D000914:Antibodies, Viral; D000956:Antigens, Viral; D015483:HIV Antibodies; D011991:Receptors, Virus; D011420:Propiolactone