Characterization of the insulin A-chain major immunogenic determinant presented by MHC class II I-Ad molecules.
Data are presented which demonstrate the minimal insulin peptide required to activate a large group of insulin-specific T hybrids following presentation by either live or fixed APC, is the N-terminal insulin-A(1-13) peptide. Functional activation and competition assays using both live and fixed APC with 19 synthesized variants of the N-terminal bovine insulin A-chain molecule permitted classification of peptide residues into MHC agretope and T cell epitope regions. Our findings indicate insulin A-chain peptide occupies the Ag binding groove of class II MHC in an extended conformation as a result of intracellular reduction of A-loop disulfide bonds. Insulin A-chain Cys7 and Cys11 residues represent two independent T cell epitopes N- and C-terminal to the A-loop region. Data are presented that demonstrate the unique residues associated with several insulin isoform molecules contribute to the peptide agretope region. Our findings may suggest peptide agretopes may subtly modify the peptide/MHC conformation presented to TCR.
Williams
D B
DB
Samuel Lunenfeld Research Institute Mount Sinai Hospital, Department of Immunology, University of Toronto, Ontario, Canada.
Ferguson
J
J
Gariepy
J
J
McKay
D
D
Teng
Y T
YT
Iwasaki
S
S
Hozumi
N
N
D000595:Amino Acid Sequence; D000818:Animals; D000938:Antigen-Presenting Cells / Q000502:physiology; D002478:Cells, Cultured; D003198:Computer Simulation; D000939:Epitopes; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D007328:Insulin / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*; D013329:Structure-Activity Relationship
D000939:Epitopes; D000949:Histocompatibility Antigens Class II; D007328:Insulin; D010446:Peptide Fragments