gd IEL migration and epithelial interactions in intestinal disease

描述(申请人提供)：γδ上皮内淋巴细胞(IEL)位于肠上皮单层内，沿着基底膜与上皮基底面接触，或位于上皮细胞之间的横向细胞间隙和紧密连接下方。γδIEL已被证明可根据具体情况预防或加重肠道疾病，但γδIEL的行为以及与肠粘膜中其他类型细胞的相互作用在很大程度上尚不明确。到目前为止，γδIEL被认为是固着的，但我的初步数据表明，γδIEL表现出动态的迁移行为，导致与上皮的广泛相互作用。此外，我已经证明γδIEL的运动依赖于上皮细胞和γδIEL对紧密连接蛋白occludin的表达，并且某些细胞因子在体外和体内都改变了这种动态的迁移行为。总之，这些观察结果导致了一个中心假设，即黏膜内稳态的破坏会对依赖闭塞素的γδIEL迁移产生不利影响，从而导致疾病的恶化。本应用的目的是1)阐明γδIEL迁移过程中γδIEL与上皮细胞相互作用的分子机制，2)确定γδIEL迁移在IL-15介导的小肠疾病中的作用，以及3)确定γδIEL迁移如何有助于保护或加重结肠炎。结合体内共聚焦显微镜、传统粘膜免疫学和上皮细胞生物学技术，将为γδIEL迁移和随后的上皮细胞相互作用在疾病过程中的贡献提供新的机制见解。此外，这些研究将加深我们对γδIEL如何在粘膜微环境中发挥作用以调节健康和疾病发展的理解，并最终可能导致旨在调节γδIEL运动的新的治疗方法，作为治疗肠道炎症的手段。这些研究是实现我的长期目标的第一步，我的长期目标是确定上皮/免疫细胞的相互作用，并确定这些相互作用与IBD和乳糜泻的相关性。拟议的研究和职业发展计划，以及我的导师、咨询委员会和芝加哥大学鼓励的跨学科方法，将提供必要的支持和额外培训，以成为学术研究环境中的独立研究人员。

DESCRIPTION (provided by applicant): γδ intraepithelial lymphocytes (IELs) are located within the intestinal epithelial monolayer, either in contact with the epithelial basal surface along the basement membrane, or in the lateral intercellular space between epithelial cells and below the tight junction. γδ IELs have been shown to prevent or exacerbate intestinal disease depending on the context, but γδ IEL behavior and interactions with other cell types in the intestinal mucosa are largely undefined. Until now, γδ IELs were presumed to be sessile, yet my preliminary data demonstrate that γδ IELs exhibit dynamic migratory behavior resulting in extensive interactions with the epithelium. Furthermore, I have shown that γδ IEL motility is dependent upon epithelial and γδ IEL expression of the tight junction protein occludin, and that certain cytokines alter this dynamic migratory behavior both in vitro and in vivo. Together, these observations have led to the central hypothesis that disruption of mucosal homeostasis adversely affects occludin-dependent γδ IEL migration leading to the exacerbation of disease. The aims of this application are to 1) elucidate the molecular mechanisms by which γδ IELs and epithelial cells interact during γδ IEL migration, 2) determine the effect of γδ IEL migration in IL-15 mediated small intestinal disease and 3) determine how γδ IEL migration contributes to the protection or exacerbation of colitis. A combination of in vivo confocal microscopy, traditional mucosal immunology, and epithelial cell biology techniques will be used to provide new mechanistic insights into the contribution of γδ IEL migration and subsequent epithelial interactions during disease. Moreover, these studies will enhance our understanding of how γδ IELs function within the mucosal microenvironment to regulate health and disease development, and may ultimately lead to new therapeutic approaches designed to modulate γδ IEL motility as means to treat intestinal inflammation. These studies are the first step toward achievement of my long-term goal to identify epithelial/immune cell interactions and determine the relevance of these interactions to IBD and celiac disease. The proposed research and career development plan, along with my mentors, advisory committee and the interdisciplinary approaches encouraged at the University of Chicago, will provide the support and additional training necessary to become an independent investigator in an academic research environment.