A reliable and reproducible method for determining specific reactivity to herpes simplex virus types 1 and 2 (HSV-1 and HSV-2) in human sera has been developed. Competition with well-characterized murine monoclonal antibodies and immunodetection of gD truncations revealed that this antibody recognizes the group Ib antigenic site of glycoprotein D, a highly conserved and protective type-common determinant. We found that treatment of either pgD-1 or pgD-2 with endo-beta-N-acetylglucosaminidase H (Endo H) generated three polypeptides which migrated more rapidly than pgD on gradient sodium dodecyl sulfate-polyacrylamide gels. (1) Thirty subclones were isolated and classified into three phenotypes as to CSA expression: (i) in CSA-positive type (20% of the clones isolated), the number of CSA-positive cells increased soon (5 hr) after seeding at 37 degree; (ii) in CSA-inducible type (33% of the clones), the number of CSA-positive cells increased after treatment with actinomycin D (ACT-D, 2 micrograms/ml), but not without ACT-D; (iii) CSA-negative type (47% of the clones), the number of CSA-positive cells did not increase after seeding or after ACT-D treatment. The envelope of HSV contains several glycoproteins: one component at 59K and a complex of two or three components at 130K, none of which corresponds in molecular weight to gp52. Using the antisera as immunological probes, we performed pulse-chase experiments with [(35)S]methionine-labeled HSV-1-infected cells and followed the disposition of the glycoproteins during the infectious cycle. Each antiserum immunoprecipitated a (35)S-labeled 52K protein from lysates of cells pulse-labeled at 5 h after infection.
Three methods were employed to determine the size of the attached oligosaccharides. The results suggest that gp52 is a precursor of gp59 and that the latter corresponds in molecular weight to one of the major glycoproteins of the virion envelope.