Role of COPII Vesicles in T cell immunity

项目概要/摘要 T 细胞在保护性适应性免疫反应中发挥重要作用，但也有助于炎症 类风湿性关节炎、I 型糖尿病和移植物排斥等疾病。他们引发了广泛的 通过分泌影响其他免疫细胞的各种细胞因子和细胞毒性因子来发挥效应活性 周围组织。然而，我们对导致这些问题的共同路径的理解仍然存在差距。 释放他们。传统上，分泌蛋白从内质网 (ER) 传播到高尔基体 囊泡中的装置由一组称为外壳蛋白复合物 II (COPII) 的蛋白质产生。而COPII 囊泡在 ER 到高尔基体的运输中发挥着关键作用，这一过程由 COPII 囊泡在 T 细胞功能中驱动 例如效应细胞因子的释放是未知的。该提案的总体目标是研究 COPII 的作用 T 细胞生物学中的囊泡，并检查它们在体内致病性和保护性 T 细胞过程中的相关性。 鉴于 COPII 囊泡在分泌途径中的基本作用，该提案的中心假设 COPII囊泡形成的破坏将导致T细胞释放特定细胞因子的缺陷 细胞，从而影响其生物功能。这个假设是根据我掌握的初步数据得出的 使用缺乏 COPII 必要成分的新型 T 细胞特异性条件敲除小鼠产生 外套，第 23 节。该提案的科学目标是：（1）确定 COPII 中断的影响 成熟幼稚 T 细胞功能的形成，以及 (2) 确定 T 细胞中 Sec23 旁系同源物的机制 功能。在第一个目标下，我将探讨废除 COPII 形成对 T 细胞的影响 保护和致病功能。为此，我将使用 T 细胞介导的成熟体内模型 抗病毒反应和同种免疫，并探索与人类 T 细胞的相关性。在第二个目标下，我 将探索 T 细胞是否依赖于不同的 COPII 旁系同源物来实现独特的功能。人类和小鼠携带 两个 Sec23 旁系同源物，据信它们可以在功能上相互补偿并表达 由于基因表达的进化变化，以组织特异性的方式。然而，我的初步数据显示 小鼠 T 细胞表达两种旁系同源物这一令人惊讶的观察结果。我将评估这两位的贡献 Sec23 旁系同源物使用现代蛋白质定量方法和广泛的细胞因子分泌模式 开始探索 COPII 介导的 T 细胞转运机制。该提案完成后，将 为 T 细胞介导的免疫提供新颖且基本的见解。它还将作为一个平台 实现我在实验免疫学方面的培训目标，并为我的长期工作提供必要的培训 作为一名医师科学家的长期目标。

PROJECT SUMMARY / ABSTRACT T cells play an important role in protective adaptive immune responses, but also contribute to inflammatory diseases such as rheumatoid arthritis, Type I diabetes mellitus, and graft rejection. They elicit a wide range of effector activities by secreting various cytokines and cytotoxic factors that influence other immune cells and surrounding tissue. However, gaps remain in our understanding of the shared common pathways that lead up to their release. Classically, secreted proteins travel from the endoplasmic reticulum (ER) to the Golgi apparatus in vesicles generated by a group of proteins called Coat Protein Complex II (COPII). While COPII vesicles play a critical role in ER-to-Golgi transport, the processes driven by COPII vesicles in T cell functions such as effector cytokine release are unknown. The overall goal of this proposal is to study the role of COPII vesicles in T cell biology, and to examine their relevance in pathogenic and protective T cell processes in vivo. Given the fundamental role of COPII vesicles in the secretory pathway, the central hypothesis of this proposal is that the disruption of COPII vesicle formation will lead to defects in the release of specific cytokines by T cells and thus impact their biological functions. This hypothesis was formed based on preliminary data I have generated using novel T cell-specific conditional knock-out mice lacking a necessary component of the COPII coat, Sec23. The scientific aims of the proposal are: (1) to determine the impact of disruption of COPII formation on mature naïve T cell functions, and (2) to determine the mechanisms of Sec23 paralogs in T cell functions. Under the first aim, I will explore the consequences of abrogating COPII formation on T cell protective and pathogenic functions. To do this, I will use well-established in vivo models of T cell-mediated anti-viral responses and alloimmunity, as well as explore relevance to human T cells. Under the second aim, I will explore whether T cells depend on different COPII paralogs for unique functions. Humans and mice carry two Sec23 paralogs, and it is believed that they can functionally compensate for each other and are expressed in a tissue-specific manner due to evolutionary shifts in gene expression. However, my preliminary data show the surprising observation that murine T cells express both paralogs. I will assess the contribution of these two Sec23 paralogs to broad cytokine secretion patterns using modern approaches to protein quantification and begin to explore the mechanisms of COPII-mediated transport in T cells. This proposal, when completed, will provide novel and fundamental insights into T cell-mediated immunity. It will also serve as a platform to achieve my training goals in experimental immunology, and to provide me with requisite training for my long- term goals as a physician scientist.