title: Specificity and detection of insulin-reactive CD4+ T cells in type 1 diabetes in the nonobese diabetic (NOD) mouse.
abstract: In the nonobese diabetic (NOD) mouse model of type 1 diabetes (T1D), an insulin peptide (B:9-23) is a major target for pathogenic CD4(+) T cells. However, there is no consensus on the relative importance of the various positions or "registers" this peptide can take when bound in the groove of the NOD MHCII molecule, IA(g7). This has hindered structural studies and the tracking of the relevant T cells in vivo with fluorescent peptide-MHCII tetramers. Using mutated B:9-23 peptides and methods for trapping the peptide in particular registers, we show that most, if not all, NOD CD4(+) T cells react to B:9-23 bound in low-affinity register 3. However, these T cells can be divided into two types depending on whether their response is improved or inhibited by substituting a glycine for the B:21 glutamic acid at the p8 position of the peptide. On the basis of these findings, we constructed a set of fluorescent insulin-IA(g7) tetramers that bind to most insulin-specific T-cell clones tested. A mixture of these tetramers detected a high frequency of B:9-23-reactive CD4(+) T cells in the pancreases of prediabetic NOD mice. Our data are consistent with the idea that, within the pancreas, unique processing of insulin generates truncated peptides that lack or contain the B:21 glutamic acid. In the thymus, the absence of this type of processing combined with the low affinity of B:9-23 binding to IA(g7) in register 3 may explain the escape of insulin-specific CD4(+) T cells from the mechanisms that usually eliminate self-reactive T cells.
authors.affiliation: Howard Hughes Medical Institute, National Jewish Health, Denver, CO 80206, USA.
authors.forename: Frances
authors.initials: F
authors.lastname: Crawford
authors.forename: Brian
authors.initials: B
authors.lastname: Stadinski
authors.forename: Niyun
authors.initials: N
authors.lastname: Jin
authors.forename: Aaron
authors.initials: A
authors.lastname: Michels
authors.forename: Maki
authors.initials: M
authors.lastname: Nakayama
authors.forename: Philip
authors.initials: P
authors.lastname: Pratt
authors.forename: Philippa
authors.initials: P
authors.lastname: Marrack
authors.forename: George
authors.initials: G
authors.lastname: Eisenbarth
authors.forename: John W
authors.initials: JW
authors.lastname: Kappler
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016367:Baculoviridae; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D000949:Histocompatibility Antigens Class II / Q000276:immunology* / Q000378:metabolism; D006825:Hybridomas / Q000276:immunology; D007328:Insulin / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D016688:Mice, Inbred NOD; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D011485:Protein Binding; D013950:Thymus Gland / Q000276:immunology
chemical_list: D000949:Histocompatibility Antigens Class II; C407099:I-A g7 antigen; D007328:Insulin; D010446:Peptide Fragments; C097877:insulin B (9-23)