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我中识别自我。病毒的检测和消除 微生物感染对人类的生存至关重要。免疫系统面临的一个挑战是感染 胞内微生物，因为微生物掩盖了它在宿主细胞中的存在。免疫的一种策略 检测微生物的系统是对不同种类的抗原进行采样，如肽、脂类和 糖脂，通过抗原递呈分子。免疫系统的一个根本独特的手臂是MR1，它 是一种抗原提呈分子，在细胞内广泛表达，在组织中广泛表达，并检测到小分子 来自微生物代谢的分子。这些特征表明，MR1已准备好检测细胞内 微生物。MR1为受MR1限制的T细胞提供抗原。这些T细胞在肺部非常普遍，而且 有能力杀死受感染的细胞。因为MR1呈递小分子抗原并采用 细胞内分布，控制细胞内环境的MR1采样的机制是不同的 来自其他抗原递呈分子。我们的假设是内体循环和内体钙 信号是MR1依赖的抗原提呈的关键，这些特征区分了MR1和MHC- 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， 已经确定了对MR1抗原重要的钙敏感转运蛋白和钙通道 演示文稿。在目标1中，我们将确定内体运输蛋白Synaxin 16在MR1中的作用。 从属抗原呈递。在目标2中，我们将定义钙敏感型突触素在MR1- 从属抗原呈递。在目标3中，我们将确定与此相关的重要钙通道 并考察了双孔通道的作用。这项研究计划是详细培训和 申请者的职业发展计划。除了实践基础科学研究外，重点是 技术，如细胞内钙测量和活细胞荧光显微镜，申请人 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 发展合作的科学会议。掌握所有这些技能将带来定义以下方面的专业知识 细胞内信号级联在抗原提呈细胞中，目标是在年底之前获得R01 K08奖。首席导师在培训科学家方面有很长的记录，指导委员会将 跟踪申请者的进步，并为申请者作为科学家的发展做出贡献。最后， 赞助机构俄勒冈健康与科学大学拥有所有必要的设施和支持 支持这项提议。

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.