title: Localization and characterization of serologic epitopes on HLA-A2.
abstract: A panel of cells expressing 68 different mutant HLA-A2 genes was generated by site-directed mutagenesis and DNA-mediated gene transfer in order to define the regions of class I MHC molecules that contribute to the epitopes recognized by mAb. Each of the variant HLA-A2 molecules differed from HLA-A2.1 by a single amino acid substitution. The substitutions were located in both the alpha-helices and beta-strands of the alpha 1 and alpha 2 domains, and included residues that are highly polymorphic and that are conserved. All but five of the variant HLA-A2 molecules were expressed at levels that ranged from approximately 25%-100% the levels found for HLA-A2.1. The remaining five variants had no detectable expression and all involved substitutions at highly conserved residues. Eleven mAbs with specificities that ranged from highly HLA-A2 specific to monomorphic were analyzed for their ability to bind the variant HLA-A2 molecules. The results demonstrate that the binding of five of 11 mAbs could be mapped to the alpha 1 and alpha 2 domains. MA2.1 was the only antibody mapped to the alpha 1 domain. CR11-351 and A2,A28M1 recognized an overlapping epitope at the amino terminal end of the alpha 2-helix, and PA2.1 and BB7.2 recognized an overlapping epitope that includes the carboxy terminus of the alpha 2-helix and a turn on one of the underlying beta-strands. These results demonstrate that positions located on the surface of the molecule, but not within the peptide-binding cleft of the molecule, are important in serological specificities.
authors.affiliation: Department of Microbiology, Medical College of Wisconsin, Milwaukee 53226.
authors.forename: K T
authors.initials: KT
authors.lastname: Hogan
authors.forename: S L
authors.initials: SL
authors.lastname: Brown
mesh_terms: D003429:Cross Reactions; D000939:Epitopes / Q000032:analysis*; D015789:HLA-A2 Antigen / Q000235:genetics / Q000276:immunology*; D006801:Humans; D009154:Mutation; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: D000939:Epitopes; D015789:HLA-A2 Antigen