Immunobiology of Corneal Antigen-Presenting Cells

项目摘要 本提案的总体目标是为主要研究者（PI）提供经验和技能 成为一名独立的研究人员，专注于研究免疫细胞的运输 机制等我的长期职业目标是建立一个全面而独立的研究计划 致力于解剖炎症和感染性角膜疾病中免疫细胞的运输机制， 从而产生新的治疗靶点。角膜抗原呈递细胞（APC）在角膜移植中具有关键作用。 保持角膜的清晰度并保持视力。APC介导的免疫应答 需要APC和T细胞之间的接触依赖性信息交换。Ag介绍人 成熟DC向幼稚T细胞诱导T细胞活化并增殖为效应T细胞，这在免疫应答中起重要作用。 转被认为是由几种机制控制，包括调节性T细胞（Treg）的作用。 APC的职业决策是APC和T细胞表面的调节分子。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 它们需要离开角膜，仍然在很大程度上未被探索。在项目前期工作中，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，其中APC亚群 表达不同的基因编码的荧光标签，并产生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， 交通信号，引导他们正常和发炎的角膜，并将用于解决以下问题 三个具体目标：1.）为了剖析APC向正常和炎症转移的分子机制， 角膜; 2.）分析APC从角膜向局部淋巴迁移的机制 在正常条件下和炎症期间的节点;和3）检查空间，时间和 角膜移植术后供、宿主APC的行为特征及其相互作用 淋巴结中的T细胞。这一目标还将分析效应T细胞或T细胞如何进入角膜。 拟议的实验将产生一个全面的，面向机制的调查， 正常条件下不同APC亚群之间的行为相似性和差异， 炎症

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.