Functional sites on the A alpha-chain. Polymorphic residues involved in antigen presentation to insulin-specific, Ab alpha:Ak beta-restricted T cells.
The interaction between the clonally selected TCR, the processed Ag peptide and the Ia molecule is not fully understood in molecular terms. Our study intended to delineate the residues of Ab alpha molecules that function as contact sites for Ag and for the TCR of a panel of T cells specific for the A chain of insulin in combination with mixed haplotype Ab alpha:Ak beta molecules. Multiple L cell transfectants expressing alpha,beta-heterodimers composed of wild-type A beta- and chimeric or mutant A alpha-chains served as antigen presenting cells. The recombinant A alpha-chains had been generated by an exchange of allelically hypervariable regions (ahv) or amino acids. The results point out a broad spectrum of b sequence requirements for the bovine insulin-specific activation of the various T cell populations. Activation of some T cells seemed quite permissive, requiring b-haplotype amino acids in any one of the three ahv, while others had strict requirements, demanding b-haplotype sequence in all three ahv. Our data stress the role of ahvII and especially ahvIII in T cell activation. Interestingly, single amino-acid substitutions in ahvII or ahvIII of Ak alpha were sufficient to bring up full stimulation potential for two T cell hybridomas. We also found that some ahv permutations influenced the Ag preference (beef insulin versus pig insulin) of some T cells. These data suggest a critical role for the three-dimensional structure of the complex formed by Ia and the processed Ag peptide. The stability of the trimolecular complex essential for T cell activation is envisioned as being the sum of the interactions between Ag/I-A, TCR/Ag, and TCR/I-A, each variable in strength and compensated for by the others.
Reske-Kunz
A B
AB
Institut für Immunologie, Johannes Gutenberg-Universität, Mainz, FRG.
Landais
D
D
Peccoud
J
J
Benoist
C
C
Mathis
D
D
D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D002417:Cattle; D002999:Clone Cells / Q000145:classification / Q000276:immunology; D000939:Epitopes / Q000276:immunology*; D006239:Haplotypes; D000949:Histocompatibility Antigens Class II / Q000235:genetics* / Q000276:immunology; D006736:Horses; D006825:Hybridomas / Q000145:classification / Q000276:immunology; D007328:Insulin / Q000235:genetics / Q000276:immunology*; D007739:L Cells / Q000276:immunology; D008213:Lymphocyte Activation; D051379:Mice; D009154:Mutation; D011110:Polymorphism, Genetic; D011994:Recombinant Proteins / Q000276:immunology; D012756:Sheep; D013045:Species Specificity; D013552:Swine; D013601:T-Lymphocytes / Q000145:classification / Q000276:immunology*; D014162:Transfection
D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007328:Insulin; D011994:Recombinant Proteins