INVARIANT T CELLS AND IL4 AND TYPE I DIABETES

不变 T 细胞与 IL4 和 I 型糖尿病

• 批准号: 2887920
• 负责人: David A. Hafler
• 金额: $ 30.86万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-30 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2887920
• 关键词: T cell receptor; T lymphocyte; calcineurin; corticosteroids; dexamethasone; diabetes mellitus genetics; enzyme activity; enzyme inhibitors; gene induction /repression; human genetic material tag; human tissue; immunogenetics; immunopharmacology; insulin dependent diabetes mellitus; interleukin 4; pharmacogenetics; phosphatidylinositol 3 kinase; tissue /cell culture; transcription factor

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.

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阻止糖尿病前期个体 发展成糖尿病 本提案将研究IL-4的调节 在T细胞的一个子集中，这与人类疾病的进展有关。 I型糖尿病 在人类疾病中， 同卵双胞胎之间的进展，都有自身反应性T细胞， 自身抗体表明，其他因素有助于发病率， 糖尿病的进展。 研究人员最近描述了一个子集， 调节性V-α 24 J-α Q T细胞的频率降低， Th 1样（缺乏IL-4分泌）在糖尿病双胞胎中的比较， 非糖尿病双胞胎 此外，疾病的非或缓慢进展者 在循环中有高水平的细胞因子IL-4和γ-IFN; 这可能反映了这些个体的更多的Th 0-或Th 2-样表型。 研究人员与研究T的实验室建立了合作关系。 细胞信号事件来解决这个问题，什么信号事件导致 IL-4分泌的损失，在不变的V-α 24 J-α Q T细胞中， 患有糖尿病的人。 核因子加塔-3的表达， c-maf、NIP 45和NF-AT物质，已被鉴定为IL-4 将通过北方印迹在以下条件下定量 激活和外源因子的条件，使细胞偏向Th 1 或Th 2表型。 这种方法将涉及转染c-maf和 将加塔-3导入不分泌IL-4的V-α 24 J-α Q T细胞中，以确定 存在阻止IL-4分泌的显性负信号。 此外，本发明还 涉及酪氨酸-641上STAT 6磷酸化的近端信号通路 将检查V-α 24 J-α Q T细胞中的细胞。 此外，由于 是糖尿病患者的V-α 24 J-alphaQ T细胞中Ca（2+）通量的缺陷， 不分泌IL-4，将调查早期TCR相关信号事件。 除了检查靶向途径外，更广泛的方法， 基因，其可以调节来自人的V-alpha 24 J-alphaQ T细胞中的IL-4表达。 将使用糖尿病受试者。 减法与PCR联用 (RDA)并进行DNA微芯片阵列杂交。 最后， 是否存在来自人的V-α 24 J-α Q T细胞的细胞因子分泌表型， 糖尿病患者可以使用产生的数据改变为Th 0/Th 2表型 从前三个目标将被审查。 的最终目标 一个项目是开发一种可以用于糖尿病前期患者的治疗方法 防止他们发展成糖尿病。