Evidence for an association between the T cell receptor/CD3 antigen complex and the CD5 antigen in human T lymphocytes

Abstract

In this work we report that CD5, a T cell accessory activation antigen and receptor for the B cell surface protein CD72, is associated with the T cell antigen receptor (TcR)/CD3 complex in human T lymphocytes. In vitro phosphorylation of either CD3 or CD5 immunoprecipitates prepared from CD3‐stimulated Jurkat and peripheral blood T cells in the presence of the detergent polyoxyethylene 10 oleyl ether (Brij96) showed, unexpectedly, an identical pattern of five phosphopolypeptides of 70, 59, 56, 21 and 18 kDa, respectively. Peptide mapping of the five bands demonstrated that the same protein kinase substrates co‐precipitated with both CD3 and CD5 and that the majority of the protein phosphorylation occurred on tyrosine residues. These data suggested that the TcR/CD3 complex and and the CD5 antigen might be associated in T cells. Evidence to support this hypothesis was obtained from analysis of immunoprecipitates prepared from surface‐iodinated T cells. Bands characteristic of the TcR and CD3 antigens were identified in CD5 immunoprecipitates and conversely, CD5 was identified in CD3 immunoprecipitates. Conformation that CD3 and CD5 co‐precipitated in the presence of Brij96 was obtained by Western blotting. Quantitative immunodepletion demonstrated that between 10 % –20 % of cell surface CD5 was associated with the TcR/CD3 complex in Brij96 detergent lysates of human T cells and, furthermore, that this association was independent of T cell activation. The association of these two receptors provides a possible physical basis for the accessory role of the CD5 antigen in T cell activation.