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Islet amyloid polypeptide is a target antigen for diabetogenic CD4+ T cells.


ABSTRACT: OBJECTIVE:To investigate autoantigens in ?-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 ?-cell antigens, we applied sequential size exclusion chromatography and reverse-phase high-performance liquid chromatography to membrane preparations of ?-cell tumors. The presence of antigen was monitored by measuring the interferon-? 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.

SUBMITTER: Delong T 

PROVIDER: S-EPMC3161333 | biostudies-literature | 2011 Sep

REPOSITORIES: biostudies-literature

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Islet amyloid polypeptide is a target antigen for diabetogenic CD4+ T cells.

Delong Thomas T   Baker Rocky L RL   Reisdorph Nichole N   Reisdorph Richard R   Powell Roger L RL   Armstrong Michael M   Barbour Gene G   Bradley Brenda B   Haskins Kathryn K  

Diabetes 20110706 9


<h4>Objective</h4>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.<h4>Research design and methods</h4>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 pr  ...[more]

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