title: A single amino acid substitution in the H-2Kb molecule generates a defined allogeneic epitope.
abstract: Using Mitomycin C mutagenesis and negative and positive selection with monoclonal antibodies specific for H-2Kb and H-2Kbm10, respectively, a mutant cell line clone, Mitc-182, was isolated. Direct sequencing of uncloned cDNA as well as PCR based cloning and sequencing of the H-2Kb182 transcript from this mutant revealed a single G-->T transversion resulting in the substitution of Trp167 by cysteine. Serologically, the mutant Kb182 and Kbm10 are almost identical as each has lost at least five Kb specific mAb epitopes and gained several new epitopes. Interestingly, the mutant cell line, Mitc-182, is efficiently recognized by alloreactive CTLs raised in reciprocal combinations, e.g. CB6 anti Cbm10 and Cbm10 anti CB6, indicating that Kb182 contains both Kb and Kbm10 specific epitopes. The mutation has not affected the ability of Kb182 to present Kb restricted antigenic peptides of Sendai and vesicular stomatitis viruses. In addition to underscoring the importance of amino acid residue 167 in alloreactivity, these results indicate a positive correlation between the gain of both an mAb epitope and a defined alloreactive CTL epitope.
authors.affiliation: Howard Hughes Medical Institute, Rockefeller University, New York, NY 10021.
authors.forename: K V
authors.initials: KV
authors.lastname: Kesari
authors.forename: G
authors.initials: G
authors.lastname: Van Bleek
authors.forename: S G
authors.initials: SG
authors.lastname: Nathenson
authors.forename: J
authors.initials: J
authors.lastname: Geliebter
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D017951:Antigen Presentation; D000956:Antigens, Viral / Q000276:immunology; D001483:Base Sequence; D000939:Epitopes / Q000235:genetics*; D006183:H-2 Antigens / Q000235:genetics*; D007518:Isoantibodies; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D009154:Mutation; D010222:Parainfluenza Virus 1, Human / Q000276:immunology; D012641:Selection, Genetic; D013602:T-Lymphocytes, Cytotoxic
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D000939:Epitopes; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D007518:Isoantibodies