Immunogenic and antigenic epitopes of immunoglobulins. II. Antigenic differences between secreted and membrane IgG demonstrated using monoclonal antibodies.
Twenty-three monoclonal antibodies with specificity for epitopes in the Fc fragment of IgG have been used to investigate antigenic differences between secreted and membrane forms of IgG produced by 2 human B lymphoblastoid cell lines (LCL). All of the monoclonals reacted with IgG secreted by the cell lines, as demonstrated by their ability to agglutinate SRBC coated with immunoglobulin isolated from culture supernatants. Membrane IgG expression was studied using direct and indirect rosette assays with antibody-coated ORBC. A surprisingly high number of antibodies, 13 on EB2 and 9 on EB4, did not bind to the cell surface immunoglobulin. These included antibodies with specificities for both C gamma 3 and C gamma 2 domain determinants. Similar results were obtained with an indirect radiobinding assay, indicating that negative results with the rosette test were not due to steric hindrance by the red cell carrier. Their performance in indirect hemagglutination indicated that most of the antibodies that did not bind to membrane IgG were of high avidity. It is concluded that the epitopes for which these antibodies are specific are not available on the cell surface. Possible explanations for the apparent antigenic differences between secreted and membrane forms of IgG are discussed against the background of previous work on the structure and mode of insertion of cell surface immunoglobulin.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000921:Antibody-Producing Cells / Q000378:metabolism*; D001666:Binding Sites, Antibody; D002460:Cell Line; D000939:Epitopes; D006386:Hemagglutination Tests; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011947:Receptors, Antigen, B-Cell / Q000276:immunology*; D012397:Rosette Formation
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007074:Immunoglobulin G; D011947:Receptors, Antigen, B-Cell