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

• 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
• 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

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）：这一修订的竞争性更新的主要目标是开发1型糖尿病的抗原特异性预防疗法，该疗法可应用于人类。我们迄今为止的研究旨在从机理上了解NOD小鼠模型中的致病过程，该模型集中于8类蛋白酪氨酸磷酸酶家族，该家族是人类体液自身免疫的主要靶点。我们已经确定了保守的抗原肽序列，这是人类和小鼠CD 4 + T细胞克隆的共同目标，和糖尿病自身抗体识别的构象表位，已经通过超过3亿年的进化保守。我们已经实施了新的技术，在体外评估T细胞细胞因子反应（ELISPOT），和方法学快速筛选致病性T细胞受体使用逆转录小鼠，以进一步评估这些抗原的致病性的重要性。该基金还支持了对人类锌转运蛋白8的体液和细胞介导的免疫反应的初步表征，这是一种主要的新的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。目前的建议，同时建立在这些初步数据体现了重点的转变，通过使用NOD小鼠作为临床前模型，用于开发基于机械的新疗法，用于预防人类1型糖尿病。具体目的1：NOD小鼠中识别ZnT 8的自发产生的自身反应性T细胞的表征。我们的方法在这里主要是经验性的，旨在通过NOD小鼠模型中的逆转录来鉴定多种候选抗原特异性TCR并评估其疾病潜力。基本目标是从ZnT 8中鉴定5-10种肽，这些肽是疾病发病机制的关键，并且可能具有治疗潜力。我们还将测试在上一个资助期内确定的phogrin TCR的糖尿病发生。具体目标2：在NOD小鼠中进行ZnT 8定向干预的临床前研究该目的探索了基于ZnT 8和phogrin的疗法的实际发展，同时注意到这种抑制可能在疾病的早期和晚期发生的机制。在实验上，这涉及分析缺乏ZnT 8的NOD小鼠中T1 D的发病率和进展速率，以及基于显性ZnT 8或发蛋白T细胞表位肽或编码cDNA的基于肽和DNA的疫苗接种策略的开发。我们预计，我们的研究结果将证实这两种自身抗原都可以成为NOD小鼠糖尿病早期和晚期的有效治疗靶点，并将为最终可用于人类的药物和方案的设计提供重要的新见解。