NOVEL IMMUNOMODULATORY STRATEGIES FOR PREVENTION AND TREATMENT OF DIABETES

鸡传染性支气管炎免疫发病机制的细胞和分子特征 胰岛素依赖型糖尿病(IDDM)是理性的 预防和治疗糖尿病的特定疗法的设计 糖尿病。然而，成功的免疫疗法预防IDDM 授权确定参与该事件的相关靶抗原(S) 自身免疫过程。这项当前的提议试图探索几部小说 自身免疫性疾病的防治策略 糖尿病在NOD小鼠身上。这项建议有三个主要目的： 首先，非肥胖糖尿病(NOD)小鼠的IDDM模型将 纵向探讨了自发丧失对两个主要 在分子水平上推测为自身抗原。我们将利用重叠 谷氨酸脱羧酶-65(GAD)和胰岛素多肽测定 胰岛来源T细胞的免疫原性和特异性。分子 T细胞分泌的细胞因子mRNA转录水平分析 对自身抗原的反应将获得关于T细胞的明确数据 自身免疫的发生和发展所必需的辅助细胞亚群 糖尿病。此外，我们建议评估糖尿病的潜在致病作用。 GAD和胰岛素多肽反应性胰岛T细胞的研究 对NOD SCID/SCID受者的采用转移实验。最后，我们 建议使用基因治疗的新技术来表达 一种免疫调节蛋白，可改善血管易损性 胰岛导致反复的自身免疫破坏。 在特定的目标2中，我们建议检查胸腺内的潜力 接种骨髓和胰岛细胞以“拯救”破坏 NOD小鼠体内残留的胰腺b细胞。而且，这些接种剂 是否也会被用来促进特定捐赠者对 长期糖尿病NOD小鼠的胸腺外胰岛移植。此外， 我们建议建立一个大型动物模型来进行临床前评估。 胸腺介导的人类免疫耐受的潜力。 在特定目标3中，白细胞跨内皮细胞迁移的机制 以及它们在白细胞向胰岛募集中的作用 将对环境进行研究。我们建议评估阻止的可能性 血小板内皮细胞黏附分子-1(PECAM-1)治疗急性心肌梗死 糖尿病的发生过程。我们还试图研究CD4-的机制- 介导的信号转导及其在细胞活化中的作用 自身抗原特异性T细胞。通过利用一种新型的CD4循环类似物， 我们寻求通过以下方法治疗和预防自身免疫性糖尿病的发病 以抗原特异性的方式阻断自身免疫反应。此外， 我们将利用从特定目标1获得的信息来创建 新型糖基化多肽，具有更高的稳定性和 多肽特异性抑制自身抗原反应性T细胞的潜能 举止。

Cellular and molecular characterization of the immune pathogenesis of insulin-dependent diabetes mellitus (IDDM) is necessary for the rational design of specific therapies for the prevention and treatment of diabetes. However, successful immunotherapy for the prevention of IDDM mandates identification of the relevant target antigen(s) involved in the autoimmune process. This current proposal seeks to explore several novel therapeutic strategies for the prevention and treatment of autoimmune diabetes in the NOD mouse. There are three major aims of this proposal: First, the non-obese diabetic (NOD) murine model of IDDM will be longitudinally probed for a spontaneous loss of tolerance to two major putative autoantigens at a molecular level. We will utilize overlapping glutamic acid decarboxylase-65 (GAD) and insulin peptides to determine the immunogenicity and specificity of islet-derived T cells. Molecular analysis of cytokine mRNA transcript levels elaborated by T cells responsive to autoantigens will elicit definitive data regarding the T helper subset requisite for the onset and progression of autoimmune diabetes. Furthermore, we propose to evaluate the diabetogenic potential of GAD and insulin peptide-reactive islet-derived T cells by performing adoptive transfer experiments to NOD scid/scid recipients. Finally, we propose to use the novel technique of gene therapy to express immunomodulatory proteins which can ameliorate the vulnerability of islets to recurrent autoimmune destruction. In specific aim 2, we propose to examine the potential of intrathymic inocula of bone marrow and islet cells for "rescue" from the destruction of residual pancreatic b-cells in the NOD mouse. Moreover, these inocula will also e used to promote donor-specific unresponsiveness to extrathymic islet allografts in long-term diabetic NOD mice. Furthermore, we propose to develop a large animal model for the preclinical evaluation of the potential of thymus-mediated immune tolerance in man. In specific aim 3, the mechanisms of leukocyte transendothelial migration and their role in the recruitment of leukocytes to the pancreatic islet milieu will be studied. We propose to assess the potential of blocking platelet endothelial cell adhesion molecule-1 (PECAM-1) to treat the diabetogenic process. We also seek to examine the mechanisms of CD4- mediated signal transduction and its role in the activation of autoantigen-specific T cells. By utilizing a novel CD4 cyclic analogue, we seek to both treat and prevent the onset of autoimmune diabetes by blocking autoimmune responses in an antigen-specific manner. Furthermore, we will utilize the information obtained from Specific Aim 1 to create novel glycosylated peptides which have increased stability and the potential to suppress autoantigen-reactive T cells in an peptide-specific manner.