Structure Peptide/Conserved T-cell Receptor Determining Diabetes

描述（由申请人提供）： 我们的数据表明，胰岛素B链氨基酸9至23肽（胰岛素B：9-23）可能是NOD小鼠自发性糖尿病的起始所必需的主要靶标，并且识别该肽的T细胞具有特异性共享保守的Valpha和Jalpha片段的T细胞受体。我将使用移植物来定义胰岛素B：9-23在胰岛自身免疫发展中的作用的具体时间和位置。我还将直接测试基因组编码的T细胞受体链对NOD小鼠的疾病和胰岛素B：9-23肽的T细胞库的识别至关重要的假设。我建议建立NOD小鼠具有假定的“不相关”的单一Jalpha 56片段，其具有与保守的Jalpha片段不同的序列（53/42），并评估它们的抗胰岛自身免疫性的发展。建立仅携带Jalpha 56的C57 BL/6小鼠，其中使用单个Jalpha来产生免疫库，并且我们将使用速度同源技术来产生同源Jalpha 56 NOD小鼠。我预测，如果Jalpha 53/42保守序列是关键的，抗胰岛素/胰岛自身免疫将不会发展为NOD小鼠只有“无关”Jalpha 56序列。如果预防发生在小鼠与单一的Jalpha 56，我们将分子创造小鼠与单一的Jalpha 56序列修改为Jalpha 53序列，我预测恢复自身免疫。对于独立的阶段，我将定义保守的α链，赋予抗胰岛素自身免疫与新的分子逆转录技术的序列。生产具有原始和修饰的α链T细胞受体的逆转录小鼠将使我们能够定义关键序列。我们将从抗B：9-23致糖尿病克隆的保守的Valpha和Jalpha分子转移氨基酸盒到“不相关的”对照α链中，以评估我们是否可以产生抗胰岛素自身免疫，并将从对照α链转移氨基酸盒到抗B：9-23致糖尿病α链中，以抑制或消除自身免疫。如果这一途径在NOD小鼠中至关重要，它将刺激寻找类似的人类关键途径。作为一个长期目标，计划包括首先在小鼠模型中开发方法，并最终将类似的逆转录和分子技术应用于来自人胰岛浸润的人T细胞受体。

DESCRIPTION (provided by applicant): Our data indicates that insulin B chain amino acids 9 to 23 peptide (insulin B:9-23) may be a primary target essential for the initiation of spontaneous diabetes in NOD mice and that T cells recognizing this peptide have a T cell receptor that specifically shares conserved Valpha and Jalpha segments. I will define using transplants, the specific timing and location of the contribution of insulin B:9-23 in the development of islet autoimmunity. I will also directly test the hypothesis that genomically encoded T cell receptor chains are crucial for disease in the NOD mouse and for recognition by the T cell repertoire of the insulin B:9-23 peptide. I propose to create NOD mice having the presumed "irrelevant" single Jalpha 56 segment, which has different sequence from the conserved Jalpha segment (53/42), and to evaluate them for development of anti-islet autoimmunity. C57BL/6 mice bearing only Jalpha 56 were established where a single Jalpha is used to create the immune repertoire, and we will create congenic Jalpha 56 NOD mice using speed congenic techniques. I predict if the Jalpha 53/42 conserved sequences are critical, anti-insulin/islet autoimmunity will not develop for NOD mice having only the "irrelevant" Jalpha 56 sequence. If prevention occurs in mice with the single Jalpha 56, we will molecularly create mice with the single Jalpha 56 sequence modified to be a Jalpha 53 sequence, and I predict restoration of autoimmunity. For the independent phase, I will define sequences of the conserved alpha chain that confer anti-insulin autoimmunity with novel molecular retrogenic techniques. Producing retrogenic mice with the original and modified alpha chain T cell receptors will allow us to define crucial sequences. We will molecularly transfer amino acid cassettes from the conserved Valpha and Jalpha of anti-B:9-23 diabetogenic clones into a "irrelevant" control alpha chain to assess whether we can create anti-insulin autoimmunity, and will transfer amino acid cassettes from control alpha chain into anti-B:9-23 diabetogenic alpha chains to suppress or abrogate autoimmunity. If this pathway is critical in the NOD mouse, it will stimulate a search for an analogous human critical pathway. As a long-term goal, plans are included to first develop methodology in the mouse model and eventually apply similar retrogenic and molecular techniques to T cell receptors of man from pancreatic islet infiltration of man.