IDENTIFICATION OF BETA CELL AUTOANTIGENS RECOGNIZED BY CD8+ T CELLS

该项目将确定MHC I类限制性β细胞 自身抗原是参与免疫应答的CD 8 + T细胞的靶标。 在IDDM的NOD小鼠模型中启动自身免疫应答。 研究开始于与大卫塞雷兹和德里罗彭尼安合作， 杰克逊实验室已经证明，从胰岛浸润的淋巴细胞 的NOD小鼠发展糖尿病可以在体外培养时喂养 胰岛细胞已从批量培养物中建立了CD 8+克隆（A1-4.4） 能够识别并特异性杀死β细胞其他三条线路 到目前为止已经分离出来，克隆正在建立中。这些克隆 在NOD小鼠自身免疫性胰岛炎的早期出现， 可能对引发疾病的关键靶点有反应。 该检测系统独立于与已知的 自身抗体我们建议分离和表征肽从 转化的β细胞的MHC I类分子是 这些CTL克隆通过以下步骤确定肽的序列： 微测序和串联质谱将允许鉴定 肽衍生的自身抗原，从而使我们能够 鉴定自身免疫反应性所针对的抗原。的 关于CD 8 + T细胞和自身抗原的肽靶标的信息 提供了一种理解T细胞破坏机制的方法 的β细胞，并可能允许开发用于预防的试剂 的IDDM。

This project will identify MHC class I restricted beta-cell autoantigens which are the targets of CD8+ T cells involved in the initiation of the autoimmune response in the NOD mouse model of IDDM. Studies begun in collaboration with David Serreze and Derry Roopenian at Jackson Laboratories have shown that infiltrating lymphocytes from islets of NOD mice developing diabetes can be cultivated in vitro when fed on islet cells. A CD8+ clone (A1-4.4)has been established from bulk cultures which recognize and specifically kill beta-cells. Three other lines have been isolated to date and clones are being established. These clones appear early in the course of autoimmune insulitis in the NOD mouse and presumable are reactive to critical targets in initiating the disease. This detection system is independent of biases related to known autoantibodies. We propose to isolate and characterize peptides from the MHC class I molecules of transformed beta-cells which are the targets for these CTL clones. Determining the sequence of the peptides by microsequencing and tandem mass spectrometry will allow identification of the autoantigen from which the peptides are derived, thus allowing us to identify antigens against which the autoimmune reactivity is directed. The information on peptide targets of the CD8+ T cells and the autoantigens provide an approach to understanding the mechanism for T cell destruction of beta-cells, and may allow development of reagents useful for prevention of IDDM.