Specificity of anti-carcinoembryonic antigen monoclonal antibodies and their effects on CEA-mediated adhesion.
The carcinoembryonic antigen (CEA) domain specificities of a library of monoclonal anti-CEA antibodies were determined to investigate the mechanisms of homophilic binding involved in CEA-mediated intercellular adhesion. Using an indirect immunofluorescence cell surface staining technique, the reactivities of these antibodies were tested systematically on Chinese hamster ovary cells stably transfected with constructs of CEA gene family members CEA, nonspecific cross-reacting antigen, CGM6, and biliary glycoprotein, as well as on stable Chinese hamster ovary transfectants expressing truncated CEA and CEA/nerve cell adhesion molecule chimeric proteins. Epitopes for these antibodies were thus localized on the CEA molecule as follows: monoclonal antibody groups 1, 2, and 6 react with epitopes in the N-terminal domain of CEA, whereas groups 3, 5a, 5b, and 9 react with determinants found in the internal repeating domains located in the C-terminal part of the CEA molecule. Groups 4 and 8 appear to react with a repeating epitope in the internal domains of CEA and in all CEA subfamily members. Fab fragments of monoclonal antibody group 1 block aggregation of CEA transfectant cells, indicating that a region in the N domain is involved in CEA homophilic interaction. Fab fragments of monoclonal antibody group 9 recognize an epitope dependent on the B2 and A3 domains and stimulate aggregation. These results support the recently reported CEA-CEA homophilic reciprocal binding model in which anti-parallel molecules are bound by 2 bonds between the N domain of one molecule and the A3B3 domain of the interacting partner.
Zhou
H
H
McGill Cancer Center, Montreal, Quebec, Canada.
Stanners
C P
CP
Fuks
A
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000494:pharmacology; D000918:Antibody Specificity; D016466:CHO Cells; D002272:Carcinoembryonic Antigen / Q000235:genetics / Q000276:immunology*; D002448:Cell Adhesion; D002450:Cell Communication; D006224:Cricetinae; D000939:Epitopes / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000276:immunology*; D014162:Transfection
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments