Immunobiology of Corneal Antigen-Presenting Cells

项目摘要 这项建议的总体目标是为首席调查员(PI)提供经验和技能 有必要成为一名独立研究人员，专注于研究免疫细胞贩运 机械装置。我的长期职业目标是建立一个全面和独立的研究计划 致力于剖析炎症和感染性角膜疾病中免疫细胞的转运机制， 导致了新的治疗靶点。角膜抗原提呈细胞(APC)在 保持角膜的清晰度，保护视力。APC介导的免疫反应 需要在APC和T细胞之间进行依赖接触的信息交换。AG演示文稿由 成熟的DC到幼稚的T细胞诱导T细胞活化和增殖为效应性T细胞，这是在 转弯被认为是由几种机制控制的，包括调节性T细胞(Treg)的行动。 APC所做的职业决定是APC和T细胞表面受调控的分子。尽管 它们在免疫系统中的独特位置，APC进入角膜所需的信号，以及线索 它们需要离开角膜，但在很大程度上仍未被探索。在这个项目的前期工作中，我们 开发了一种新的多光子活体显微镜(MP-IVM)模型来研究完整的APC 麻醉小鼠的角膜。这种成像方法使用转基因小鼠，在转基因小鼠中，APC亚群 表达不同的基因编码荧光标签，并制作APC的3D延时电影 亚细胞分辨率，与周围细胞相互作用。将对这些细胞进行研究，以研究 引导他们到达正常和发炎的角膜的交通信号灯，并将用于解决以下问题 三个具体目标：1.APC转化为正常和炎症的分子机制剖析 角膜；2.)分析APC从角膜向局部淋巴迁移的机制 正常情况下和炎症期间的结节；以及3)检查空间、时间和 角膜移植后供者和宿主APC的行为特征及其相互作用 淋巴结内有T细胞。这个目标还将分析效应器T细胞或Tregs如何进入角膜。 拟议的实验将产生一个全面的，以机制为导向的调查 不同APC亚群在正常和正常情况下的行为异同 发炎。

Project Summary The overall goal of this proposal is to provide the principal investigator (PI) with the experiences and skills necessary to become an independent researcher, with the focus on studying immune cell trafficking mechanisms. My long-term career goal is to establish a comprehensive and independent research program dedicated to dissect the trafficking mechanism of immune cells in inflammatory and infectious corneal disease, leading to new therapeutic targets. Corneal antigen-presenting cells (APC) have a critical role in maintaining the clarity of the cornea and preserving vision. APC-mediated immune responses require contact-dependent information exchange between APC and T cells. Ag presentation by mature DC to naive T cells induces T cell activation and proliferation into effector T cells, which is in turn thought to be controlled by several mechanisms, including the action of regulatory T cells (Treg). Career decisions taken by APC are regulated molecules on the surface of APC and T cells. Despite their unique position in the immune system, the signals APC need to enter the cornea, and the clues they require to leave the cornea, are still largely unexplored. In preliminary work for this project, we have developed a new multiphoton intravital microscopy (MP-IVM) model to study APC in intact corneas of anesthetized mice. This imaging approach uses transgenic mice, in which APC subsets expresses distinct genetically encoded fluorescent tags, and produces 3D time-lapse movies of APC at subcellular resolution, interacting with surrounding cells. These cells will be studied to investigate the traffic signals that guide them to normal and inflamed corneas and will be used to address the following three specific aims: 1.) To dissect the molecular mechanisms by which APC travel to normal and inflamed corneas; 2.) To analyze the mechanisms involved in migration of APC from the cornea to local lymph nodes under normal conditions and during inflammation; and 3) examine the spatial, temporal and behavioral characteristics of donor and host APC after corneal transplantation and their interaction with T cells in lymph nodes. This aim will also analyze how effector T cells or Tregs enter the cornea. The proposed experiments will generate a comprehensive, mechanism-oriented survey of the behavioral similarities and differences between distinct APC subsets under normal condition and inflammation.