Analysis of major histocompatibility complex class I-restricted hapten recognition by mutation of the V-J joining of T cell receptor alpha chains.

Abstract

Hapten-specific T cell responses are responsible for chemically induced immune disorders. However, the molecular details of hapten interactions with T cell receptors (TCR) are poorly understood. Recent studies of trinitrophenyl (TNP)-specific responses revealed major histocompatibility complex-associated TNP-peptides as dominant epitopes for CD8+ and CD4+ T cells. The present study is based on the observation that two H-2Kb/TNP-specific CTL clones (II/7 and III/1), differing exclusively in two amino acids of their TCR alpha chains, also differed in their carrier specificities for various TNP-peptides. The genes of the two alpha chains and the common beta chain were cloned into expression vectors. Transfection of the TCR alpha chain of clone III/1 into a hybridoma of clone II/7 also transferred the fine specificity of clone III/1, indicating that the small alpha chain variations were indeed responsible for the different carrier specificities. Point mutations bridging the difference between the alpha chains of clones II/7 and III/1 and functional studies of the respective TCR alpha beta transfectants into a TCR-negative hybridoma revealed an unexpected result: the two receptors did not represent examples of structural complementarity for different sets of hapten-peptide conjugates; rather, they resembled two structures of principally similar specificity but of significantly different overall affinity. This was demonstrated more directly by comparing the fine specificities of III/1 transfectants expressing or not expressing the co-receptor CD8: the CD8-negative III/1 transfectant assumed a specificity pattern indistinguishable from that of a CD8-expressing, II/7-derived transfectant. Hence, comparable alterations of antigen recognition may be induced either by subtle TCR alterations or by removal of CD8, i.e. by the presence or absence of a non-polymorphic adhesion molecule.