Epitope analysis of human cytomegalovirus glycoprotein complexes using murine monoclonal antibodies.
A panel of 10 monoclonal antibodies reactive with human cytomegalovirus (HCMV) glycoproteins was generated. These antibodies immunoprecipitated disulfide-linked complexes which contained glycoproteins with molecular weights of 130,000, 93,000, and 52,000. These complexes were designated gC-I. Epitope analysis of gC-I was done with a simultaneous two antibody binding assay. A network of epitopes was revealed which clustered in three major domains designated I, II, and III. Antibodies within individual domains I and II showed strong mutual inhibition of each other's binding. However, there were multiple antibody interactions between domains I and II. For example, the binding of most antibodies in domain I was augmented to some extent by antibodies from domain II. However, the binding of only one antibody from domain II was augmented by all antibodies from domain I. The augmentation in binding between two antibodies was dependent on the native structure of gC-I and was sensitive to conformational changes due to nonionic detergent extraction of gC-I and/or disruption of disulfide bonds. A synergistic effect was also observed between antibodies in domains I and II in a virus neutralization assay. A neutralizing antibody had a much greater neutralizing activity in the presence of a nonneutralizing antibody, which also enhanced the binding of the neutralizing antibody in the simultaneous two antibody binding assay. Also, two antibodies which were nonneutralizing individually were neutralizing when used in combination. Such antibodies also augmented each other's binding in the simultaneous two antibody binding assay. Finally, domain III consisted of a nonneutralizing antibody that inhibited the binding of all antibodies in domains I and II. This antibody also inhibited the neutralizing activity of a neutralizing antibody in a virus neutralizing assay.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D003587:Cytomegalovirus / Q000276:immunology*; D000939:Epitopes; D007163:Immunosorbent Techniques; D008562:Membrane Glycoproteins / Q000276:immunology; D009500:Neutralization Tests; D011487:Protein Conformation; D014759:Viral Envelope Proteins / Q000276:immunology*; D010948:Viral Plaque Assay
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins