INVARIANT T CELLS AND IL4 AND TYPE I DIABETES

描述(改编自申请人的摘要)：本申请的目的 建议是确定哪些因素可以阻止糖尿病前期患者 患上糖尿病。这项提案将调查IL-4的调节 在T细胞的一个子集中，这些T细胞与人类的发展有关 I型糖尿病。在人类疾病中，疾病的不完全整合 同卵双胞胎之间的进展，既有自身反应性T细胞，也有 自身抗体表明，其他因素也与发病率和 IDDM的进展。调查人员最近描述了一组 调节性V-α24J-αQ T细胞频率降低， 糖尿病双生子Th1样(缺乏IL-4分泌)与 非糖尿病双胞胎。此外，疾病的非进展或缓慢进展者 循环中细胞因子IL-4和γ-干扰素水平较高； 这可能反映了这些个体更多的Th0或Th2样表型。 调查人员与调查T细胞的实验室建立了合作关系 细胞信令事件来解决这个问题，信令事件会导致什么 不变V-α24J-αQ T细胞IL-4分泌缺失的研究 糖尿病患者。核因子GATA-3的表达， C-MAF、NIP45和NF-AT物种，已被鉴定为IL-4 转录因子，将在条件下用Northern印迹定量 激活和外源因子使细胞偏向Th1的条件 或Th2表型。这一方法将涉及c-maf和c-maf的转染。 GATA-3作用于非IL-4分泌的V-α24J-αQ T细胞 有一个显性的负信号阻止IL-4的分泌。此外， 涉及Tyr-641上STAT6磷酸化的近端信号通路 在V-alpha24J-alphaQ T细胞将被检查。此外，由于在那里 糖尿病患者V-α24J-αQ T细胞钙(2)通量缺陷 不分泌IL-4，早期TCR相关信号事件将被调查。 除了检查目标路径之外，还有更广泛的方法来隔离目标 可能调节V-α24J-αQ T细胞IL-4表达的基因 糖尿病的受试者，将被使用。减法与聚合酶链式反应联用 将进行(RDA)和DNA微芯片阵列杂交。最后， V-α24J-αQ T细胞分泌细胞因子的表型 可以使用生成的数据将糖尿病患者改变为Th0/Th2表型 从前三个目标开始，我们将对其进行检查。的最终目标是 该项目是开发一种可用于糖尿病前期患者的治疗方法 以防止他们发展为糖尿病患者。

DESCRIPTION (adapted from applicant's abstract): The intent of this proposal is to determine what factors prevent pre-diabetic individuals from developing diabetes. This proposal will investigate the regulation of IL-4 in a subset of T cells, which are implicated in the progression of human type I diabetes. In human disease, incomplete concordance of disease progression between identical twins, both with autoreactive T cells and autoantibodies suggests that other factors contribute to the incidence and progression of IDDM. Investigators have recently described a subset of regulatory V-alpha 24J-alphaQ T cells that are reduced in frequency and are Th1-like (lacking IL-4 secretion) in diabetic twins as compared to non-diabetic twins. In addition, the non- or slow-progressors to disease had high levels of the cytokines IL-4 and gamma-IFN in their circulation; this may reflect a more Th0- or Th2-like phenotype for these individuals. Investigator has set up collaborations with laboratories investigating T cell signaling events to address the question, what signaling events lead to the loss of IL-4 secretion in the invariant V-alpha 24J-alphaQ T cells in individuals with diabetes. Expression of the nuclear factors, GATA-3, c-maf, NIP45, and NF-AT species, that have been identified as IL-4 transcription factors, will be quantitated by Northern blot under conditions of activation and conditions of exogenous factors to bias cells toward a Th1 or Th2 phenotype. This approach will involve transfection of c-maf and GATA-3 into non IL-4 secreting V-alpha 24J-alphaQ T cells to determine if there is a dominant negative signal preventing IL-4 secretion. Further, proximal signaling pathways involving phosphorylation of STAT 6 on tyr-641 in the V-alpha 24J-alphaQ T cells will be examined. In addition, as there are defects in Ca(2+) flux in V-alpha 24J-alphaQ T cells from diabetics that do not secrete IL-4, early TCR associated signaling events will be surveyed. Besides examining targeted pathways, broader approaches, to isolate target genes, which may regulate IL-4 expression in V-alpha 24J-alphaQ T cells from subjects with diabetes, will be used. Subtraction methods coupled with PCR (RDA) and DNA microchip array hybridization will be performed. Lastly, whether the cytokine secretion phenotype of V-alpha 24J-alphaQ T cells from the diabetics can be altered to a Th0/Th2 phenotype using the data generated from the first three aims will be examined. The eventual goal of the project is to develop a therapy that can be administered to pre-diabetics that prevent their progression into diabetics.