Cloning of molecular targets of Cell Mediated Autoimmunity in Type 1 Diabetes

1 型糖尿病细胞介导的自身免疫分子靶标的克隆

基本信息

批准号：
8233962
负责人：
HOWARD W DAVIDSON
金额：
$ 32.72万
依托单位：
UNIVERSITY OF COLORADO DENVER
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-12-14 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8233962
关键词：
Animal Model Animals Antigens Appearance Attention Autoantibodies Autoantigens Autoimmune Diseases Autoimmunity Biological Markers Breeding CD4 Positive T Lymphocytes CD8B1 gene Cells Child Chronic Disease Clinical Clinical Research Clinical Trials Cloning Complementary DNA Conduct Clinical Trials DNA Data Development Diabetes Mellitus Diabetes autoantibodies Disease Drug Design Elderly Epitopes Evaluation Evolution Family Funding Goals Grant Haplotypes Health Human Immunity Immunodominant Epitopes In Vitro Inbred NOD Mice Incidence Individual Insulin Insulin-Dependent Diabetes Mellitus Intervention Knowledge Laboratories Mediating Methodology Modeling Molecular Target Mus NOD/SCID mouse Organ PTPRN gene Pancreas Pathogenesis Peptides Phase Population Pre-Clinical Model Prevention Prevention therapy Process Protein Tyrosine Phosphatase Protocols documentation Reagent Regulatory T-Lymphocyte Research Role Screening procedure Specificity Staging System T cell response T-Cell Receptor T-Lymphocyte T-Lymphocyte Epitopes TNFRSF10A gene Testing Therapeutic Time To autoantigen Translating Vaccination autoreactive T cell base cell mediated immune response cytokine diabetic disease natural history enzyme linked immunospot assay high risk human subject insight man member model development mouse model new technology novel therapeutic intervention preclinical study protein aminoacid sequence research study response therapeutic target vaccination strategy young adult

项目摘要

DESCRIPTION (provided by applicant): The major goal of this revised competing renewal is the development of antigen-specific preventative therapies of type 1 diabetes that can be translated to man. Our studies to date have been aimed at a mechanistic understanding of the pathogenic processes in the NODmouse model focused on a family of class 8 protein tyrosine phosphatases that are major targets of humoral autoimmunity in humans. We have identified conserved antigenic peptide sequences that are common targets of both human and mouse CD4+ T cell clones, and conformational epitopes recognized by diabetes autoantibodies that have been conserved through over 300 million years of evolution. We have implemented new technologies for the in vitro evaluation of T cell cytokine responses (ELISPOT), and methodologies for rapidly screening pathogenic T cell receptors using retrogenic mice to further evaluate the pathogenic importance of these antigens. The grant also supported the initial characterization of humoral and cell-mediated immune responses to human Zn transporter 8, a major new ss-cell specific human diabetes autoantigen; the first to be discovered in over a decade. The current proposal while building upon these preliminary data embodies a shift in emphasis by using the NOD mouse as a pre-clinical model for the development of mechanistically-based new therapies for prevention of type 1 diabetes in humans. Specific Aim 1: Characterization of spontaneously-arising autoreactive T cells recognizing ZnT8 in NOD mice. Our approach here is largely empirical aimed at identifying multiple candidate antigen-specific TCRs and evaluating their disease potential by retrogenesis in a NOD mouse model. The fundamental objective is to identify 5-10 peptides from ZnT8 that are key to the pathogenesis of disease and which may have therapeutic potential. We will also test the diabetogenesis of the phogrin TCRs identified in the previous funding period. Specific aim 2: Pre-clinical studies of ZnT8-directed interventions in NOD mice This aim explores the practical development of ZnT8-and phogrin-based therapies, while at the same time giving heed to the mechanisms by which such suppression might occur in early and late stages of the disease. Experimentally this involves analysis of the incidence and rate of progression of T1D in NOD mice lacking ZnT8, and development of peptide and DNA based vaccination strategies based on dominant ZnT8 or phogrin T cell epitope peptides, or encoding cDNA. We anticipate that our results will confirm that both autoantigens can be effective therapeutic targets in NOD mice, both early and late during diabetogenesis, and will provide important new insight into the design of drugs and protocols that can ultimately be translated to humans.

描述（由申请人提供）：本次修订后的竞争性更新的主要目标是开发可应用于人类的 1 型糖尿病抗原特异性预防疗法。迄今为止，我们的研究旨在从机制上理解 NODmouse 模型中的致病过程，重点关注 8 类蛋白酪氨酸磷酸酶家族，这些酶是人类体液自身免疫的主要靶标。我们已经鉴定出作为人类和小鼠 CD4+ T 细胞克隆的共同靶点的保守抗原肽序列，以及糖尿病自身抗体识别的构象表位，这些构象表位在 3 亿多年的进化过程中一直保守。我们已经实施了体外评估 T 细胞细胞因子反应 (ELISPOT) 的新技术，以及使用逆转录小鼠快速筛选致病性 T 细胞受体的方法，以进一步评估这些抗原的致病重要性。该资助还支持对人锌转运蛋白 8 的体液和细胞介导的免疫反应的初步表征，锌转运蛋白 8 是一种主要的新型 ss 细胞特异性人类糖尿病自身抗原；这是十多年来首次发现的。目前的提议在这些初步数据的基础上体现了重点的转变，即使用 NOD 小鼠作为临床前模型，开发基于机械的新疗法来预防人类 1 型糖尿病。具体目标 1：NOD 小鼠中自发产生的识别 ZnT8 的自身反应性 T 细胞的表征。我们在这里的方法主要是经验性的，旨在识别多个候选抗原特异性 TCR，并通过在 NOD 小鼠模型中逆发生来评估其疾病潜力。基本目标是从 ZnT8 中鉴定出 5-10 种肽，这些肽对于疾病的发病机制至关重要，并且可能具有治疗潜力。我们还将测试上一个资助期间确定的噬蛋白 TCR 的糖尿病发病机制。具体目标 2：针对 NOD 小鼠的 ZnT8 定向干预的临床前研究该目标探索基于 ZnT8 和噬菌蛋白的疗法的实际开发，同时关注这种抑制在早期和晚期可能发生的机制。疾病的晚期阶段。实验上，这涉及分析缺乏 ZnT8 的 NOD 小鼠中 T1D 的发病率和进展速度，以及基于显性 ZnT8 或噬菌蛋白 T 细胞表位肽或编码 cDNA 的肽和 DNA 疫苗接种策略的开发。我们预计我们的结果将证实这两种自身抗原在糖尿病发生的早期和晚期都可以成为 NOD 小鼠的有效治疗靶点，并将为最终可转化为人类的药物和方案的设计提供重要的新见解。