Islet Amyloid Polypeptide Is a Target Antigen for Diabetogenic CD4+ T Cells
Islet Amyloid Polypeptide Is a Target Antigen for Diabetogenic CD4+ T Cells
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DOI：
10.2337/db11-0288
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发表时间：
2011-09-01
期刊：
影响因子：
7.7
通讯作者：
Haskins, Kathryn
中科院分区：
文献类型：
--
作者：
Debug, Thomas;Baker, Rocky L.;Haskins, Kathryn
为了研究β细胞中的自身抗原，我们使用了一组来自NOD小鼠的致病性T细胞克隆。我们在这项研究中特别关注的是高度致糖尿病的T细胞克隆BDC-5.2.9的靶抗原的鉴定。研究设计和方法-为了纯化β细胞抗原，我们应用连续尺寸排阻色谱法和反相高效液相色谱法制备β细胞肿瘤的膜。通过测量在NOD抗原呈递细胞存在下响应于色谱级分的BDC-5.2.9的干扰素-γ产生来监测抗原的存在。峰抗原组分进行了分析，离子阱质谱法，并通过肽分析和候选蛋白进行了进一步研究，在可能的情况下，测试胰岛组织基因敲除mice. MTS-质谱分析显示存在的胰岛淀粉样多肽（IAPP）的抗原成分。IAPP缺陷小鼠的胰岛不能在体外和体内刺激BDC-5.2.9，并且存在强烈刺激BCD-5.2.9的IAPP衍生肽，证实了IAPP作为抗原靶点。结论-IAPP是致糖尿病CD 4 T细胞克隆BDC-5.2.9的靶抗原。糖尿病60：2325-2330，2011
OBJECTIVE-To investigate autoantigens in beta-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 beta-cell antigens, we applied sequential size exclusion chromatography and reverse-phase high-performance liquid chromatography to membrane preparations of beta-cell tumors. The presence of antigen was monitored by measuring the interferon-gamma 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. Diabetes 60:2325-2330, 2011