Characterization of the antigen-specific T cell receptor from a pigeon cytochrome c-specific T cell hybrid.
The monoclonal antibody, A2B4-2, specifically bound to and inhibited antigen-induced IL-2 release from the cloned pigeon cytochrome c-specific T cell hybrid, 2B4. The initial immunoprecipitation with these reagents demonstrated a protein of 85-90kd on non-reducing conditions, and two bands of 45-50 and 40-44 on reducing conditions. These data enabled us to conclude that this monoclonal antibody bound to the antigen receptor on the 2B4 cell. In this paper we have presented results that further define the receptor structure. The migration pattern of the protein on SDS-PAGE in the presence and absence of reducing agents was consistent with the interpretation that the receptor has intrachain disulfide bonds that create globular domains. Sequence data from the recently described cDNA clones that encode receptor genes confirm that there are cysteine residues in positions that could be involved in such disulfide bonding. Since both alpha and beta chains behave identically on the SDS-PAGE we predict that both chains have the same double domain structure. The antigen receptor has a heterodimeric structure. A relatively simple acidic alpha chain is bound to one of two forms of the beta chain. Some of the receptors have a beta chain equal in molecular weight to the alpha chain, while some of the beta chains (beta') are heavier and more acidic. The difference in beta chain forms appears to be due to different levels of glycosylation of this chain. The cDNA sequence data demonstrate that there are several possible carbohydrate addition sites on the protein encoded by this gene so it may be that 2B4 beta chains are present that are either completely or partially glycosylated at these sites. Finally, we have presented a preliminary experiment that indicates that a smaller 20-25kd molecule is associated with the antigen receptor. The well characterized T3 molecule appears to be in a complex with the clonotypic structure on human T cells. The crosslinking data that we present suggests that a similar molecule may be present in the murine system.
D000818:Animals; D000911:Antibodies, Monoclonal; D003001:Cloning, Molecular; D010856:Columbidae; D003574:Cytochrome c Group / Q000032:analysis*; D004247:DNA / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D006822:Hybrid Cells / Q000276:immunology*; D008970:Molecular Weight; D011948:Receptors, Antigen, T-Cell / Q000032:analysis* / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003574:Cytochrome c Group; D000939:Epitopes; D011948:Receptors, Antigen, T-Cell; D004247:DNA