T-Cell Recognition of Ia Molecules Selectively Altered by a Single Amino Acid Substitution

T lymphocytes recognize foreign antigen together with allele-specific determinants on membrane-bound class I and class II (Ia) gene products of the major histocompatibility complex. To identify amino acids of class II molecules critical to this recognition process, the genes encoding the β chains of the I-A^k molecule were cloned from a wild-type B-cell hybridoma and from an immunoselected variant subline showing distinct serological and T-cell stimulatory properties. Nucleotide sequencing and DNA-mediated gene transfer established that a single base transition (G --> A) encoding a change from glutamic acid to lysine at position 67 in the I-A_β^k molecule accounted for all the observed phenotypic changes of the variant cells. These results confirm the importance of residues 62 to 78 in the amino terminal domain of I-A_β for class II--restricted T-cell recognition of antigen and demonstrate the ability of a single substitution in this region to alter this recognition event.