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  • Title: Islet amyloid polypeptide is a target antigen for diabetogenic CD4+ T cells.
    Author: Delong T, Baker RL, Reisdorph N, Reisdorph R, Powell RL, Armstrong M, Barbour G, Bradley B, Haskins K.
    Journal: Diabetes; 2011 Sep; 60(9):2325-30. PubMed ID: 21734016.
    Abstract:
    OBJECTIVE: To investigate autoantigens in β-cells, we have used a panel of pathogenic T-cell clones that were derived from the NOD mouse. Our particular focus in this study was on the identification of the target antigen for the highly diabetogenic T-cell clone BDC-5.2.9. RESEARCH DESIGN AND METHODS: To purify β-cell antigens, we applied sequential size exclusion chromatography and reverse-phase high-performance liquid chromatography to membrane preparations of β-cell tumors. The presence of antigen was monitored by measuring the interferon-γ production of BDC-5.2.9 in response to chromatographic fractions in the presence of NOD antigen-presenting cells. Peak antigenic fractions were analyzed by ion-trap mass spectrometry, and candidate proteins were further investigated through peptide analysis and, where possible, testing of islet tissue from gene knockout mice. RESULTS: Mass-spectrometric analysis revealed the presence of islet amyloid polypeptide (IAPP) in antigen-containing fractions. Confirmation of IAPP as the antigen target was demonstrated by the inability of islets from IAPP-deficient mice to stimulate BDC-5.2.9 in vitro and in vivo and by the existence of an IAPP-derived peptide that strongly stimulates BCD-5.2.9. CONCLUSIONS: IAPP is the target antigen for the diabetogenic CD4 T-cell clone BDC-5.2.9.
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