T cell hybridomas of (B10 X B10.BR)F1 genotype with reactivity to bovine insulin (BI) were established to analyze the restriction and antigen fine specificity of (H-2b X H-2k)F1 T cells towards BI. Our data indicate a focusing of the response on two epitopes on the insulin molecule, the A chain loop determinant comprising amino acids A8 and A10, as well as the glutamic acid residue in position 4 of the A chain. Both were recognized either separately or in conjunction. Unexpectedly, the T cell hybridomas exhibited a marked preference for recognizing insulin in the context of F1-unique restriction elements of Ab alpha Ak beta type rather than parental high-responder I-Ab molecules. Analysis of the response of primed lymph node T cells of (B10 X B10.BR)F1 mice towards BI corroborated the finding of a preponderant corecognition of F1-unique I-A molecules.