Formation, Positioning, Motility, and Function of Tissue Resident Memory CD8+ T cells After Influenza Infection

项目总结/摘要-项目3 组织驻留记忆T细胞（TRM）是在组织中建立的非再循环CD 8 + T细胞。 感染后的外周组织。当再次遇到相同或相关的病原体时，这些记忆T 细胞迅速重新激活，并提供直接效应功能，这可能意味着生命和 在致命挑战模型中死亡。它们倾向于对保守抗原具有特异性，并且在 流感，可能是实现更广泛的交叉反应和通用疫苗的解决方案的一部分。 了解它们是如何被监管的，它们如何介导最佳保护，以及它们是如何建立的， 这是该项目的关键目标。除了用于鉴定T细胞类型的标志物（CD 3，CD 8， CD 44、CD 62 L、CD 69），几种其他细胞表面标志物用于鉴定记忆T细胞亚群。 组织中当与整合素β-1配对形成VLA-1时，CD 49 a是细胞外基质中胶原蛋白的受体。 基质，并且是我们在2004年首次用于定义组织中这些细胞的原型TRM标记。CD103， 当与β-7整联蛋白配对时，结合上皮细胞间连接处表达的E-钙粘蛋白。这些 TRM的“标志物”对它们的建立和功能至关重要，但很少有人确定它们在TRM中的作用。 CD 49 a和CD 103的体内功能。我们建议测试与这些粘附力中的每一个如何相关的假设 分子的作用是将记忆T细胞定位在不同的解剖位置，调节与细胞的通讯， 上皮细胞，与功能分化相关的遗传编程有关，从而调节 CD 8 + T细胞运动性、存活率和最佳免疫保护。具体目标是： 目的1：确定决定分化、建立和维持的机制。 流感感染后的TRM亚群。 目的2：研究T细胞-上皮细胞-基质相互作用的机制， 定位在气道中。 目的3：探讨CD 49 a和CD 103在优化免疫保护中的作用。 为了实现这些目标，我们使用基因组学、流式细胞术、体外和体内模型的组合来研究 这些细胞的分化和功能。采用的核心技术是活体多光子显微镜 （IV-MPM）应用于我们实验室开发的流感气管炎活动物模型。我们扩展了这个 使用包含流感病毒的原代气道类器官培养物的创新体外系统的方法 感染和病毒特异性CD 8 T细胞，以研究调节细胞运动和T- 细胞/上皮细胞相互作用。我们的研究结果将导致新的方法来优化免疫保护 流感和减少疾病负担。

PROJECT SUMMARY/ABSTRACT – PROJECT 3 Tissue resident memory T cells (TRM) are non-recirculating CD8+ T cells that become established in peripheral tissues after an infection. Upon re-encounter with the same or related pathogen(s), these memory T cells rapidly reactivate and provide immediate effector function that can mean the difference between life and death in a lethal challenge model. They tend to be specific for conserved antigens, and in the case of influenza, could be part of the solution to achieving more broadly cross-reactive and universal vaccine. Understanding how they are regulated, how they mediate optimal protection, and how they are established and maintained are critical goals of this project. Besides the markers used to identify the T cell types (CD3, CD8, CD44, CD62L, CD69), several other cell surface markers are used to identify memory T cell subsets in the tissues. CD49a, when paired to integrin beta-1 to form VLA-1, is a receptor for collagen in the extracellular matrix and is the prototypic TRM marker first used to define these cells in the tissue by us in 2004. CD103, when paired with beta-7 integrin, binds to E-cadherin expressed in the junctions between epithelial cells. These “markers' of TRM are critical to their establishment and function, yet little has been done to determine the in vivo functions of CD49a and CD103. We propose to test hypotheses related to how each of these adhesion molecules acts to position memory T cells in different anatomical locations, regulate communication with the epithelium, are associated with genetic programming linked to functional differentiation, thereby regulating CD8+ T cell motility, survival, and optimal immune protection. The Specific Aims are: Aim 1: Determine the mechanisms that determine differentiation, establishment, and maintenance of TRM subsets after influenza infection. Aim 2: Investigate mechanisms of T cell-epithelial cell-matrix interactions required for motility and positioning in the airways. Aim 3: Determine the functions of CD49a and CD103 in optimizing immune protection. To achieve these aims, we use combinations of genomics, flow cytometry, in vitro and in vivo models to study the differentiation and function of these cells. A core technology employed is intravital multiphoton microscopy (IV-MPM) applied to a live animal model of influenza tracheitis that our lab developed. We extend this approach to an innovative in vitro system using primary airway organoid cultures incorporating influenza infection and virus-specific CD8 T cells to study fine aspects of the mechanisms regulating cell motility and T- cell/epithelial cell interactions. The results of our studies will lead to novel ways to optimize immune protection from influenza and reduce the burden of disease.