Identification of Novel Autoantigens in Type 1 Diabetes

1 型糖尿病中新型自身抗原的鉴定

基本信息

批准号：
7235736
负责人：
MATTEO G LEVISETTI
金额：
$ 0.11万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2004
资助国家：
美国
起止时间：
2004-04-01 至 2009-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7235736
关键词：
NOD mouse SCID mouse SDS polyacrylamide gel electrophoresis T lymphocyte autoantibody autoantigens cellular immunity flow cytometry insulin dependent diabetes mellitus mass spectrometry monoclonal antibody pancreatic islets polymerase chain reaction southern blotting tissue /cell culture

项目摘要

DESCRIPTION (provided by applicant): Type 1 diabetes is an autoimmune disease characterized by the progressive destruction of pancreatic beta-cells that leads to insulinopenia and hyperglycemia. In spite of extensive study, little is known about the initial beta-cell autoantigens that trigger the autoimmune response. This application focuses on identifying the antigens that the immune system targets early in the pathogenesis of diabetes. Specific Aim 1: Characterize autoantibodies to beta cell antigens and identify the antigens. We plan on producing monoclonal antibodies from NOD mice of various ages that are reactive with beta cell surface antigens. These antibodies will be characterized and then used to identify the target antigens present on the surface of the beta cells. These beta cell target antigens will be cloned, sequenced and expressed in cell lines by transfection, and in systems that will allow purification of protein for experimentation. Specific Aim 2: Characterize the T cell reactivity against autoantigens that are identified by the monoclonal antibodies. The generation of peptides, and/or protein, from identified beta-cell autoantigens will allow for the pursuit of autoreactive T cells from the spleen, peripancreatic lymph nodes and islet infiltrates of NOD mice. T cell reactivity against all beta cell autoantigens that are identified will be characterized at the level of protein and T cell peptide epitopes will be characterized. Specific Aim 3: Evaluate the pathogenic role of autoantibodies and T cells. The functional role played by any identified beta-cell antigens in the pathogenesis of diabetes, and the antibodies and T cells which target them, will be examined. The phenotype of T cells reactive with identified antigens will be explored and characterized. Together these aims may lead to an understanding of the early pathologic events in Type 1 diabetes, specifically of the beta-cell antigens that are involved, which is essential for the development of preventive and therapeutic interventions.

描述（由申请人提供）： 1 型糖尿病是一种自身免疫性疾病，其特征是胰腺 β 细胞进行性破坏，导致胰岛素减少和高血糖。尽管进行了广泛的研究，但人们对触发自身免疫反应的初始β细胞自身抗原知之甚少。该应用的重点是鉴定免疫系统在糖尿病发病早期所针对的抗原。具体目标 1：表征 β 细胞抗原的自身抗体并鉴定抗原。我们计划从不同年龄的 NOD 小鼠中生产与 β 细胞表面抗原发生反应的单克隆抗体。这些抗体将被表征，然后用于识别 β 细胞表面上存在的靶抗原。这些β细胞靶抗原将通过转染在细胞系中以及允许纯化实验用蛋白质的系统中进行克隆、测序和表达。具体目标 2：表征 T 细胞针对单克隆抗体识别的自身抗原的反应性。从已鉴定的 β 细胞自身抗原中产生肽和/或蛋白质将允许从 NOD 小鼠的脾脏、胰周淋巴结和胰岛浸润物中追踪自身反应性 T 细胞。 T 细胞针对所有已鉴定的 β 细胞自身抗原的反应性将在蛋白质水平上进行表征，并且将表征 T 细胞肽表位。具体目标 3：评估自身抗体和 T 细胞的致病作用。将检查任何已识别的 β 细胞抗原在糖尿病发病机制中所发挥的功能作用，以及针对它们的抗体和 T 细胞。将探索和表征与已识别抗原反应的 T 细胞表型。这些目标共同可能有助于了解 1 型糖尿病的早期病理事件，特别是所涉及的 β 细胞抗原，这对于制定预防和治疗干预措施至关重要。