Regulation of Human IELs by CD94 and HLA-E

描述（申请人的摘要）：在人肠道的微环境中 上皮，具有细胞溶解功能的武装效应T细胞（T-IEL）， 受CD 94/NKG 2（一个新的HLA-E特异性异二聚体家族）的严格调控 最初在NK细胞上发现的受体。这些受体可以转换 通过使用不同的激活或抑制性NKG 2开启和关闭适应性反应 同种型，并似乎调节显着大的T-IEL克隆扩增， 健康的个体。在乳糜泻中，一种与上皮细胞 损伤时，它们只表达激活同种型，这表明， 失调导致疾病。拟议研究的总体目标是 建立一个细胞溶解效应T细胞介导的适应性免疫的模型， 细胞在肠道微环境中受先天性巨噬细胞CD 94/NKG 2受体的调节。 免疫力研究人员将重点关注三个基本的具体目标， 了解这个调节系统的生物学意义。在 特异性目的1、HLA-E和G在肠上皮细胞中的表达及调控 上皮细胞，细胞因子产生的GALT在正常和炎症 条件将用于刺激HLA-E和G的表达， 隔离IEC、IEC线路和专业APC。在具体目标2中， 以及通过T-IEL调节CD 94/NKG 2同种型，研究者将 表征不同CD 94/NKG 2同种型的表达模式， 表达不同CD 94/NKG 2的T-IEL的TCR V-β和V-α库 同种型（通过光谱分析和测序），以及 表达不同CD 94/NKG 2同种型的T-IEL。在具体目标3中，功能性 由T-IEL表达的CD 94 INKG 2受体的性质，克隆的表达T-IEL的 将使用单个NKG 2同种型来确定它们如何有助于细胞溶解 功能和细胞因子分泌，并表征生化信号 涉案这项拟议中的研究将在分子定义明确的系统中进行测试 一般假设NK受体和非经典的MHC I类 分子调节T-IEL和肠上皮细胞之间的相互作用。 这些结果可能对我们理解 在高抗原性细胞中微调溶细胞性T细胞应答的过程 肠道环境，调节局部免疫和控制上皮 细胞损伤。

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.