Differential activation of T cells by T-cell receptor ligand analogs

Although CD4(+) T cell responses to protein-derived antigen have well been understood, the epitopes recognized by hapten-specific CD4(+) T cells have not been fully defined. In this study, we characterized the response of a T cell hybridoma (5D10.1B8) which is specific for a hapten, N-hydroxysuccinimidyl-4-azidobenzoate (HSAB) restricted by MHC class II I-A(d). Using three different antigen presenting cells (APCs) expressing I-A(d), the role of class II MHC proteins in haptenic antigen presentation and subsequent activation of 5D10.1B8 has been examined. Activation of 5D10.1B8 T cells by HSAB analogs was also performed. Our results show that each APC activated T cells differentially and that interleukin-2 (IL-2) augmented antigen-presenting ability of all the APCs, suggesting that increased expression of class II MHC protein by IL-2 played an important role in HSAB presentation and T cell activation. Finally, early T cell receptor-dependent signals induced by HSAB or its analogs were examined by phosphotyrosine immunoblot analysis, and showed that tyrosine phosphorylation level of a 18-20 kD protein increased upon stimulation.