MR1-dependent presentation of Mycobacterium tuberculosis: a unique role for calcium signaling

项目摘要 免疫系统的基本作用是从非自我中检测自我。检测和消除 微生物感染对人类生存至关重要。对免疫系统的一个挑战是来自 因为微生物掩盖了它在宿主细胞中的存在。免疫系统的一个策略 检测微生物的系统是对不同种类的抗原进行采样，例如肽、脂质和 糖脂，通过抗原呈递分子。免疫系统的一个基本独特的手臂是MR 1， 是一种抗原呈递分子，在细胞内，在组织中广泛表达，并检测小的 来自微生物代谢的分子。这些特征表明，MR 1有望检测细胞内 微生物MR 1将抗原呈递给MR 1限制性T细胞。这些T细胞在肺部非常普遍， 有能力杀死被感染的细胞。由于MR 1呈递小分子抗原并采用 细胞内分布，管理MR 1采样的细胞内环境的机制是不同的 从其他抗原呈递分子中分离出来我们的假设是内体再循环和内体钙 信号传导是MR 1依赖性抗原呈递的关键，这些特征将MR 1与MHC-1区分开来。 Ia抗原呈递。我们使用结核分枝杆菌（Mtb）作为细胞内感染的模型， 已经鉴定了对MR 1抗原重要的钙敏感运输蛋白和钙通道 演示文稿.在目标1中，我们将定义内体运输蛋白Syntaxin 16在MR 1 - 2中的作用。 依赖性抗原呈递在目标2中，我们将定义钙敏感性突触结合蛋白在MR 1 - 1中的作用。 依赖性抗原呈递在目标3中，我们将确定参与此过程的重要钙通道。 过程，并检查了双孔通道的作用。本研究计划是详细培训的一部分， 申请人的职业发展计划。除了动手基础科学研究，重点是 例如细胞内钙的测量和活细胞荧光显微术的技术， 我将完成科学写作和生物统计学的课程，以扩大科学知识，并将参加 科学会议，以发展合作。获得所有这些技能将导致专业知识， 在抗原呈递细胞中的细胞内信号传导级联，目的是在抗原呈递细胞的细胞内信号传导结束时获得R 01。 K 08奖。主要导师在培训科学家方面有着长期的记录，导师委员会将 跟踪申请人的进展，并有助于申请人作为科学家的发展。最后 赞助机构，俄勒冈州健康与科学大学，拥有所有必要的设施和支持， 支持这个提议。

Project Summary The fundamental role of the immune system is to detect self from non-self. The detection and elimination of microbial infection is critical for human survival. One challenge to the immune system is infection from an intracellular microbe because the microbe masks its presence in a host cell. One strategy of the immune system to detect microbes is the sampling of different kinds of antigens, such as peptides, lipids and glycolipids, by antigen presenting molecules. A fundamentally unique arm of the immune system is MR1, which is an antigen presenting molecule that is intracellular, ubiquitously expressed across tissues, and detects small molecules derived from microbial metabolism. These features suggest that MR1 is poised to detect intracellular microbes. MR1 presents antigens to MR1-restricted T cells. These T cells are highly prevalent in the lungs and have the ability to kill infected cells. Because MR1 presents small molecule antigens and adopts an intracellular distribution, the mechanisms governing MR1 sampling of the intracellular environment are distinct from other antigen presenting molecules. Our hypothesis is that endosomal recycling and endosomal calcium signaling are key for MR1-dependent antigen presentation, and that these features distinguish MR1 from MHC- Ia antigen presentation. We use Mycobacterium tuberculosis (Mtb) as a model for intracellular infection and have identified calcium-sensitive trafficking proteins and calcium channels important for MR1 antigen presentation. In Aim 1, we will define the role of the endosomal trafficking protein Syntaxin 16 in MR1- dependent antigen presentation. In Aim 2, we will define the role of calcium sensitive Synaptotagmins in MR1- dependent antigen presentation. In Aim 3, we will identify the important calcium channels involved in this process and examined the role of Two-pore channels. This research plan is part of a detailed training and career development plan for the applicant. In addition to hands-on basic science research with an emphasis on techniques such as measurement of intracellular calcium and live-cell fluorescence microscopy, the applicant will complete coursework in scientific writing and biostatistics to expand scientific knowledge and will attend scientific conferences to develop collaborations. Acquisition of all of these skills will lead to expertise in defining intracellular signaling cascades in antigen presenting cells, with the goal of obtaining an R01 by the end of the K08 award. The primary mentor has a long track record of training scientists and a Mentoring Committee will track the applicant's progress and contribute to the applicant's development as a scientist. Finally, the sponsoring institution, Oregon Health & Science University, has all the facilities and support necessary to support the proposal.