Isolation of anti-idiotypic antibodies to T cells using an anti-framework determinant antibody.
A panel of anti-idiotypic antibodies to the T cell line HPB-ALL was produced by screening with a novel enzyme-linked immunoadsorption assay (ELISA). Using the beta framework I (beta F1) monoclonal antibody directed at a common determinant on the human T cell receptor beta subunit, we were able to specifically capture the receptor molecule from a cell lysate preparation and use this as the basis of an ELISA assay. Hybridoma supernatants were tested for their ability to bind to the receptor thus captured. A total of four antibodies were isolated by this method, and they were shown to immunoprecipitate a disulfide-linked heterodimer composed of alpha (49 kDa) and beta (40 kDa) subunits from HPB-ALL cells, similar to the subunits recognized by the beta F1 antibody. Furthermore, all four antibodies blocked the binding of T40/25, an anti-idiotype to HPB-ALL. Three of these antibodies blocked the binding of anti-Leu 4 to a similar degree as did T40/25, while one did not. This suggests that these new anti-idiotypic antibodies recognize distinct but associated idiotypic determinants. The isolation of such antibodies for any particular T cell line or tumor promises to be useful for biological studies of T cell malignancy in humans.
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis / Q000276:immunology; D006825:Hybridomas / Q000378:metabolism; D007130:Immunoglobulin Idiotypes / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D011233:Precipitin Tests; D011971:Receptors, Immunologic / Q000502:physiology; D013601:T-Lymphocytes / Q000276:immunology* / Q000648:ultrastructure
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000939:Epitopes; D007130:Immunoglobulin Idiotypes; D011971:Receptors, Immunologic