Critical role of HLA-DR beta 1 residue 58 in multiple polymorphic epitopes recognized by xenogeneic and allogeneic antibodies.
In a previous study, we identified glutamic acid at position 58 in DR (beta 1*1101) as critical for the epitopes recognized by the DRw11-specific mAb GS88.2, as well as the I-LR1 mAb that recognizes a polymorphic epitope on DR(alpha,beta 1*1101) and some DP molecules. The purpose of this study was to determine whether other polymorphic residues contribute to these epitopes and whether DR beta glutamic acid or alanine 58 and DP beta glutamic acid 56, the analogous position in DP beta, contribute to epitopes recognized by other anti-class-II mAb and allosera. Site-directed mutagenesis and transfection were used to produce cells bearing wild-type or mutant class II molecules that were analyzed with mAbs by flow cytometry and with human allosera by absorption and subsequent microcytotoxicity assays. These studies demonstrate that the residue at DR beta position 58 plays a central role in at least three different mAb epitopes and an epitope recognized by anti-DRw11 allosera. Substitution of glutamic acid for alanine at position 58 of eight DR beta chains caused gain of binding of four mAbs to all of the mutant molecules, except DR(alpha,beta 4*0101). These data suggest that the side chains of DR beta 58 and DP beta 56 point outward from the alpha-helix and directly contact antibody.
D000409:Alanine / Q000276:immunology; D000595:Amino Acid Sequence; D000910:Antibodies, Heterophile / Q000276:immunology*; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002478:Cells, Cultured; D000939:Epitopes / Q000276:immunology*; D005347:Fibroblasts; D005971:Glutamates / Q000276:immunology*; D018698:Glutamic Acid; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007126:Immunoglobulin Allotypes / Q000276:immunology*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011110:Polymorphism, Genetic; D013329:Structure-Activity Relationship; D014162:Transfection
D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D000939:Epitopes; D005971:Glutamates; D006684:HLA-DR Antigens; D007126:Immunoglobulin Allotypes; D018698:Glutamic Acid; D000409:Alanine