Dual Overlapping Peptides Recognized by Insulin Peptide B:9-23 T Cell Receptor AV13S3 T Cell Clones of the NOD Mouse

J Autoimmun. 2000 May;14(3):231-7. doi: 10.1006/jaut.2000.0369.

Abstract

T cells isolated from islets of non-obese diabetic (NOD) mice are enriched for insulin-reactive cells. The great majority of these T cells recognize insulin B chain peptide (B:9-23). B:9-23 reactive T cell clones are diabetogenic and show a dramatic TCR alpha -chain restriction (predominant AV13S3). We have studied the reactivity of five different B:9-23 reactive T cell clones to truncated peptides and alanine substituted analogues of B:9-23. Amongst these AV13S3 T cell clones, one reacted with peptide B:9-16 and four with B:13-23. The two peptides have in common only four amino acids (B:13-16; EALY). Having defined minimal peptide epitopes, we evaluated a mutant insulin sequence (B:13 glutamine) which retains metabolic activity. As predicted, this single amino acid change abrogated T cell reactivity. In addition, we have created a modified I-A(g7)gene with the B:9-23 peptide covalently linked to I-A(g7). Antigen presenting cells transfected with this construct were excellent presenting cells for all clones studied. The definition of dual peptide motifs and creation of bioactive covalent I-A(g7)-B:9-23 should facilitate studies of the pathogenic significance and antigen recognition by B:9-23 reactive diabetogenic T cells.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Alanine / genetics
  • Alanine / immunology
  • Amino Acid Sequence
  • Amino Acid Substitution
  • Animals
  • Antigen Presentation
  • Epitopes, T-Lymphocyte / immunology
  • Glutamine / genetics
  • Glutamine / immunology
  • Histocompatibility Antigens Class II / genetics
  • Histocompatibility Antigens Class II / immunology
  • Insulin / genetics
  • Insulin / immunology*
  • Islets of Langerhans / cytology
  • Mice
  • Mice, Inbred NOD
  • Molecular Sequence Data
  • Peptide Fragments / genetics
  • Peptide Fragments / immunology*
  • Peptides / immunology*
  • Receptors, Antigen, T-Cell, alpha-beta / immunology*
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*

Substances

  • Epitopes, T-Lymphocyte
  • Histocompatibility Antigens Class II
  • I-A g7 antigen
  • Insulin
  • Peptide Fragments
  • Peptides
  • Receptors, Antigen, T-Cell, alpha-beta
  • insulin B (9-23)
  • Glutamine
  • Alanine