Intracellular pathogens and innate immunity

项目摘要/摘要(总体) 这项申请是一项计划项目拨款的竞争性续订，其名称为“细胞内病原体和 先天免疫力。“我们解决的中心问题是，细胞内的病原体是如何被 宿主和免疫系统如何整合多种信号以诱导适当的反应，以及 相反，病原体如何避免和/或操纵宿主反应以促进其发病。在ITS中 在之前的迭代中，该计划项目利用新出现的全基因组技术来 巨噬细胞对多种细胞内细菌病原体的转录反应 并发现，感染了多种细胞内的细菌病原体导致了刺痛的激活 下游诱导以I型表达为主的核心转录反应 干扰素调节的基因。为了更深入地探索宿主对细胞内病原体的反应，每个 P01实验室已经开始研究感染后的转录后反应。在项目1中，Portnoy 扩展了他对c-diAMP和STING在单核细胞增多性李斯特菌感染和 豁免权。他进一步建议研究单核细胞增多性李斯特氏菌如何避免自噬，以及这是如何影响的。 免疫的致病机制和诱导作用。最后，他建议将主持人和 细菌蛋白在感染过程中的泛素化。在项目2中，考克斯研究了自噬小体是如何形成的 在含有结核分枝杆菌的吞噬小体上被选择性地激活。接下来，基于他的全球 泛素化研究确定IRF7是一种促进细胞内生长的转录因子 他将阐明IRF7促进感染的机制。最后，他提议 确定结核分枝杆菌感染过程中宿主蛋白磷酸化的功能变化。在项目3中， 万斯建议通过使用CRISPR/Cas9来解开刺痛激活的相反影响 产生选择性消除特定叮人依赖的新型敲入小鼠的技术 活体内的反应。接下来，他将确定嗜肺军团菌如何针对中枢新陈代谢 调节因子mTORC1，以及这如何影响宿主对感染的反应。最后，他们将使用全面的 核心C中的图谱技术，以确定对嗜肺性乳杆菌的新的转录后反应。在核心中 巴顿将提供全面的指导，并维持小鼠的群体，繁殖小鼠，回交小鼠，并使用 CRISPR-Cas9将产生新的基因组编辑小鼠模型以确定后基因组的体内功能 上面确定的转录反应。在Core C中，Krogan将应用基于新的质谱学和 定量描述翻译后相互作用的生物信息学方法，包括泛素化和 在感染过程中发生的磷酸化。核心A的目的是确保科学进步和 通过提供科学、组织和行政领导来促进协同，这将是 在所有P01实验室及其附属小组的月度会议上通过广泛的科学审查完成。

Project Summary/Abstract (Overall) This application is a competitive renewal of a program project grant entitled, “Intracellular pathogens and innate immunity.” The central problem that we address is how intracellular pathogens are recognized by the host and how the immune system integrates multiple signals to induce an appropriate response, and conversely, how pathogens avoid and/or manipulate the host response to promote their pathogenesis. In its previous iterations, this program project took advantage of newly emerging genome-wide technologies to characterize the transcriptional response of macrophages to a variety of intracellular bacterial pathogens and discovered that infection with diverse intracellular bacterial pathogens led to the activation of STING and the downstream induction of a core transcriptional response dominated by expression of type I interferon-regulated genes. To explore the host response to intracellular pathogens more deeply, each of the P01 labs has begun to investigate post-transcriptional responses to infection. In Project 1, Portnoy extends his studies on the roles of c-di-AMP and STING during Listeria monocytogenes infection and immunity. He further proposes to examine how L. monocytogenes avoids autophagy and how this impacts both pathogenesis and induction of immunity. Finally, he proposes to characterize the role of both host and bacterial protein ubiquitylation during infection. In Project 2, Cox examines how autophagosome formation is selectively activated at the Mycobacterium tuberculosis-containing phagosome. Next, based on his global ubiquitylation studies that identified IRF7 as a transcription factor that promotes the intracellular growth of M. tuberculosis, he will elucidate the mechanisms by which IRF7 promotes infection. Finally, he proposes to identify functional changes in host protein phosphorylation during M. tuberculosis infection. In Project 3, Vance proposes to disentangle the opposing effects of STING activation by using CRISPR/Cas9 technologies to generate novel STING knock-in mice that selectively eliminate specific STING-dependent responses in vivo. Next, he will determine how Legionella pneumophila targets the central metabolic regulator mTORC1, and how this affects host responses to infection. Finally, they will use comprehensive profiling technologies in Core C to identify novel post-transcriptional responses to L. pneumophila. In Core B, Barton will provide overall direction and maintain colonies of mice, breed mice, backcross mice, and use CRISPR-Cas9 to generate novel genome-edited mouse models to determine the in vivo function of post- transcriptional responses identified above. In Core C, Krogan will apply novel mass spectrometry-based and bioinformatics approaches to quantitatively profile posttranslational interactions, including ubiquitylation and phosphorylation that occur during infection. The purpose of Core A is to ensure scientific progress and promote synergy by providing scientific, organizational, and administrative leadership, which will be accomplished by extensive scientific review during monthly meetings of all the P01 lab and affiliated groups.