Regulation of Human IELs by CD94 and HLA-E

描述(申请人摘要)：在人体肠道的微环境中 上皮臂效应T细胞(T-IEL)具有细胞溶解功能 受CD94/NKG2的严格调控--一种新的人类白细胞抗原-E特异性异二聚体家族 最初在NK细胞上发现的受体。这些受体可以切换 不同激活或抑制NKG2的适应性反应 同种类型，并似乎调节非常大的T-IEL克隆性扩张 健康的个体。在乳糜泻中，一种与上皮性病变有关的疾病 损伤，它们只表达激活的同工型，这表明 调节失调会导致疾病。拟议研究的总体目标是 建立细胞溶解效应T细胞介导的获得性免疫模型 肠道微环境中细胞受天然CD94/NKG2受体的调节 豁免权。调查员将重点关注三个基本的具体目标 了解这一调控系统的生物学意义。在……里面 特异性靶点1、人类白细胞抗原-E和G在肠道中的表达及调控 正常和炎症时GALT产生的上皮细胞和细胞因子 条件将被用来刺激新鲜人类白细胞抗原E和G的表达 隔离的IEC、IEC线路和专业APC。在具体目标2中，表述 以及T-IEL对CD94/NKG2亚型的调节，研究人员将 鉴定不同CD94/NKG2亚型的表达模式 表达CD94/NKG2的T-IEL的TCRV-β和V-α谱系 同工型(通过光谱分析和测序)，以及细胞因子谱 表达不同CD94/NKG2亚型的T-IEL。在具体目标3中，职能 T-IEL表达的CD94INKG2受体的特性及其克隆表达 单一的NKG2亚型将被用来确定它们对细胞溶解的贡献 功能和细胞因子的分泌以及生化信号的特征 牵涉其中。拟议的研究将在一个分子定义明确的系统中进行测试。 NK受体和非经典MHC-I类受体的一般假设 分子调节T-IEL和肠上皮细胞之间的相互作用。 这一结果可能会对我们理解 在高抗原中微调细胞溶解T细胞反应的过程 肠道环境，调节局部免疫，控制上皮 细胞受损。

DESCRIPTION (Applicant's Abstract): In the microenvironment of the human gut epithelium, armed effector T cells (T-IELs) with cytolytic functions are tightly regulated by CD94/NKG2, a novel family of HLA-E specific heterodimeric receptors originally identified on NK cells. These receptors can switch adaptive responses on and off by using different activating or inhibitory NKG2 isotypes and appear to regulate remarkably large T-IEL clonal expansions in healthy individuals. In celiac disease, a condition associated with epithelial damage, they exclusively express activating isotypes, suggesting that dysregulation causes disease. The overarching goal of the proposed research is to develop a model of how adaptive immunity mediated by cytolytic effector T cells is regulated in the gut microenvironment by CD94/NKG2 receptors of innate immunity. The investigator will focus on three specific aims fundamental to understanding the biological significance of this regulatory system. In Specific Aim 1, expression and regulation of HLA-E and G on intestinal epithelial cells, cytokines produced in the GALT in normal and inflammatory conditions will be used to stimulate expression of HLA-E and G by freshly isolated IECs, IEC lines and professional APCs. In Specific Aim 2, expression and regulation of CD94/NKG2 isotypes by T-IELs, the investigator will characterize the pattern of expression of the different CD94/NKG2 isotypes, the TCR V-beta and V-alpha repertoire of T-IELs expressing distinct CD94/NKG2 isotypes (by spectroanalysis and sequencing), and the cytokine profile of T-IELs expressing distinct CD94/NKG2 isotypes. In Specific Aim 3, functional properties of CD94INKG2 receptors expressed by T-IELs, cloned T-IELs expressing single NKG2 isotypes will be used to determine how they contribute to cytolytic function and cytokine secretion and to characterize the biochemical signals involved. The proposed research will test in a molecularly well-defined system the general hypothesis that NK receptors and non-classical MHC class I-like molecules regulate interactions between T-IELs and intestinal epithelial cells. The results are likely to have important implications for our understanding of the processes that fine tune cytolytic T cell responses in the high antigen environment of the gut, regulating local immunity and controlling epithelial cell damage.