Flow cytometric detection of herpes simplex virus type 2 infected and transformed cells by immunoenzymatic and by indirect immunofluorescence staining.
The glucose oxidase antiglucose oxidase (GAG) immunoenzymatic staining procedure has been used to detect herpes simplex virus (HSV) antigens microscopically. In this study, the GAG procedure was adapted to cells in suspension, and its potential usefulness in flow cytometry was examined. HSV-2 infected monkey kidney and HSV-2 transformed mouse cells were stained using antisera to HSV-2 or to an HSV-2 specific protein with a molecular weight of 38 Kd, respectively, with the GAG procedure. Flow cytometric analysis of the GAG stained cells was then performed by the measurement of scattered light intensity in the angular intervals 1 degree-2 degrees, 2.5 degrees-19 degrees, and 3 degrees-6 degrees. The greatest scattered light intensity decrement caused by staining occurred in the 3 degrees-6 degrees angular interval, as predicted by previous work. In infected cells, which stain intensely by immunofluorescence, the difference between positively and negatively stained cells was adequate for detecting infected cells using the GAG method; however, this was not the case for the lightly staining transformed cells. The indirect immunofluorescence method of analysis of the same populations was superior to the scattered light method of analysis of the GAG stained infected and transformed cells.
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D002461:Cell Line, Transformed / Q000166:cytology* / Q000382:microbiology / Q000528:radiation effects; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D005562:Formazans; D005949:Glucose Oxidase / Q000276:immunology; D000882:Haplorhini; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000166:cytology; D008027:Light; D051379:Mice; D011817:Rabbits; D012542:Scattering, Radiation; D018139:Simplexvirus / Q000276:immunology*; D013194:Staining and Labeling / Q000379:methods*; D013778:Tetrazolium Salts; D014466:Ultraviolet Rays
D000956:Antigens, Viral; D005562:Formazans; D013778:Tetrazolium Salts; D005949:Glucose Oxidase