I-Ak polymorphisms define a functionally dominant region for the presentation of hen egg lysozyme peptides.
The class II molecules of the MHC not only bind processed antigenic peptides but also interact with the TCR. This latter interaction is thought to be the basis for allele specific "restriction" of Ag presentation to T cells. The specificity of this interaction is likely due to amino acid differences in a small number of polymorphic or "hypervariable" regions located in the amino terminal domains of the alpha- and beta-chains. We have explored the functional significance of these polymorphic regions in an I-Ak-restricted, hen egg lysozyme specific Ag presentation system in which the measurement of IL-2 production by T cell hybridomas was used as the indicator of TCR recognition of the I-A/Ag complex. Chimeric I-A molecules, in which b allelic residues were substituted in one or more of the polymorphic regions of the A alpha k chain or in which d allelic residues were substituted in one or more of the polymorphic regions of the A beta k chain, were used to examine the contribution of each polymorphic region of the molecule to its function. The results obtained demonstrate that the regions between residues 69 to 76 of the A alpha k chain and the regions between residues 63 to 67 and 75 to 78 of the A beta k-chain exert a dominant effect on the presentation of lysozyme peptides by I-Ak to the T cell hybridomas in our panel. These observations were confirmed and extended by the analysis of Ag presentation by seven serologically selected mutants, all of which have amino acid interchanges in or around the dominant polymorphic regions. The results suggest that the serologically selected mutants fail to present Ag not because they fail to bind the peptide Ag but because the amino acid substitutions destabilize the interaction between the Ia/peptide complex and the TCR. Use of the recently published hypothetical model for class II structure to interpret the Ag presentation results suggests that the dominant polymorphic regions lie across from one another near one end of the alpha-helices that form the two walls of the proposed Ag-binding cleft located on the top surface of the class II molecule. Furthermore, the majority of the amino acids which have been changed in the serologically selected mutants have side chains which are postulated to point up toward the exterior of the molecule and would, therefore, be potential contact residues for the TCR.
Rosloniec
E F
EF
Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, CO 80206.
Vitez
L J
LJ
Beck
B N
BN
Buerstedde
J M
JM
McKean
D J
DJ
Landais
D
D
Benoist
C
C
Mathis
D
D
Freed
J H
JH
D000595:Amino Acid Sequence; D000818:Animals; D000938:Antigen-Presenting Cells / Q000276:immunology*; D002645:Chickens; D002678:Chimera; D004527:Egg Proteins / Q000276:immunology*; D005799:Genes, Dominant; D000949:Histocompatibility Antigens Class II / Q000235:genetics* / Q000276:immunology; D006822:Hybrid Cells / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D009113:Muramidase / Q000276:immunology*; D009154:Mutation; D010455:Peptides / Q000276:immunology*; D011110:Polymorphism, Genetic; D013601:T-Lymphocytes / Q000276:immunology; D014162:Transfection
D004527:Egg Proteins; D000949:Histocompatibility Antigens Class II; D010455:Peptides; D009113:Muramidase