Characterization of the HLA-restrictive elements of a rubella virus-specific cytotoxic T cell clone: Influence of HLA-DR4β chain residue 74 polymorphism on antigenic peptide-T cell interaction

Abstract

The influence of glutamic acid (E)-alanine (A) dimorphism at position 74 of the DR4β chain on cytotoxic T cell recognition of an antigenic rubella virus peptide, E1(273-284), was studied using a panel of B cell lines and B cell transfectants expressing different HLA-DRB1 alleles as antigen-presenting cells and targets in 51Cr-release assays. Only a cell lines expressing the DRB1(*)0403, DRB1(*)0406 or DRB1(*)0407 subtypes which shared a residue, E, at position 74 in the DR4β chain when sensitized with E1(273-284) elicited strong cytotoxic T lymphocyte responses. However, in direct binding and antibody inhibition assays, it was shown that biotinylated E1(272-285) could bind to DR molecules with residues other than E at position 74, including DRB1(*)0401, DRB1(*)0404 and DRB1(*)1101 expressed on transfectants. E1(272-285) bound with similar affinity to the transfectant with DRB1(*)0403, which has E at position 74, as well as the transfectant with DRB1(*)0404, which does not. When T-B cell engagement rates were compared in cell conjugate assays, the percentage of T-B conjugates was higher when peptide-pulsed transfectants with DRB1(*)0403 were used than with transfectants expressing DRB1(*)0404. Hence, the HLA DRβ1 polymorphism at position 74, while not critical for the binding affinity of E1(272-285) to the HLA molecule, appears to be a primary determinant of restricted recognition and subsequent activation of the peptide-specific T cells.