Constitutive and Inflammatory Immunopeptidome of Pancreatic beta-Cells
AuthorDudek, NL; Tan, CT; Gorasia, DG; Croft, NP; Illing, PT; Purcell, AW
PublisherAMER DIABETES ASSOC
University of Melbourne Author/sDudek, Nadine; Gorasia, Dhana; CROFT, NATHAN; PURCELL, ANTHONY; TAN, CHOR; ILLING, PATRICIA
AffiliationAcademic Services and Registrar
Melbourne Dental School
Biochemistry and Molecular Biology
Microbiology and Immunology
Document TypeJournal Article
CitationsDudek, N. L., Tan, C. T., Gorasia, D. G., Croft, N. P., Illing, P. T. & Purcell, A. W. (2012). Constitutive and Inflammatory Immunopeptidome of Pancreatic beta-Cells. DIABETES, 61 (11), pp.3018-3025. https://doi.org/10.2337/db11-1333.
Access StatusOpen Access
Type 1 diabetes is characterized by the autoimmune destruction of pancreatic β-cells. Recognition of major histocompatibility complex (MHC)-bound peptides is critical for both the initiation and progression of disease. In this study, MHC peptide complexes were purified from NIT-1 β-cells, interferon-γ (IFN-γ)-treated NIT-1 cells, splenic and thymic tissue of 12-week-old NOD mice, and peptides identified by mass spectrometry. In addition to global liquid chromatography-tandem mass spectrometry analysis, the targeted approach of multiple-reaction monitoring was used to quantitate the immunodominant K(d)-restricted T-cell epitope islet-specific glucose-6-phosphatase catalytic subunit-related protein (IGRP)₂₀₆₋₂₁₄. We identified >2,000 MHC-bound peptides; 1,100 of these presented by β-cells grown under normal conditions or after exposure to IFN-γ. These include sequences from a number of known autoantigens. Quantitation of IGRP₂₀₆₋₂₁₄ revealed low-level presentation by K(d) (~25 complexes/cell) on NIT-1 cells after IFN-γ treatment compared with the simultaneous presentation of the endogenously processed K(d)-restricted peptide Janus kinase-1₃₅₅₋₃₆₃ (~15,000 copies/cell). We have successfully sequenced peptides from NIT-1 β-cells under basal and inflammatory conditions. We have shown the feasibility of quantitating disease-associated peptides and provide the first direct demonstration of the disparity between presentation of a known autoantigenic epitope and a common endogenously presented peptide.
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