The allorecognition of H-2Kb-specific CD4-CD8- T cell hybridomas is influenced by the substitution at residue 256 of MHC class I molecules.
Our previous studies demonstrated that allorecognition of HTB176.10 and HTB177.2, H-2Kb-reactive CD4-CD8- T cell hybridomas is markedly influenced by the exchange of the alpha 3 domain between H-2Kb and H-2Dp. The recombinant genes of the exon 4 between H-2Kb and H-2Dp were constructed to determine the residues of the alpha 3 domain that influence the allorecognition of these T cell hybridomas. Seven recombinant genes of the exon 4 were generated by in vivo recombination in Escherichia coli. Chimeric genes containing these recombinants were transfected into L cells and the transfectants expressing equivalent amounts of chimeric molecules were selected by flow cytometry. Studies on responses of these T cell hybridomas to the chimeric molecules confirmed our previous observation that the primary structure of the alpha 3 domain influences the allorecognition by the hybridomas. Moreover, it was indicated that residue 256 on the alpha 3 domain markedly affects the allorecognition by the T cell hybridomas, although substitutions at residues 184, 193, 195, 197, 262, and 264 exerted some effects on the T cell recognition. Further studies with the use of a single amino acid mutant of H-2Kb at residue 256 confirmed the effect of substitution at residue 256 on allorecognition of the T cell hybridomas. Taken together, results of this study demonstrated that polymorphism of the alpha 3 domain is indeed involved in the formation of allodeterminants recognized by TCR.
Tanabe
M
M
Department of Immunology, Tokyo University Hospital, University of Tokyo, Japan.
Minami
M
M
Kano
K
K
Takiguchi
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D000954:Antigens, Surface / Q000235:genetics; D001483:Base Sequence; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens; D005091:Exons; D006183:H-2 Antigens / Q000235:genetics / Q000276:immunology*; D006825:Hybridomas; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000096:biosynthesis; D007739:L Cells; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D011110:Polymorphism, Genetic; D011994:Recombinant Proteins; D013329:Structure-Activity Relationship; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
D000945:Antigens, Differentiation, T-Lymphocyte; D000954:Antigens, Surface; D015704:CD4 Antigens; D016827:CD8 Antigens; D006183:H-2 Antigens; D007376:Interleukin-2; D009841:Oligonucleotides; D011994:Recombinant Proteins