Intracellular pathogens and innate immunity

项目概要/摘要（总体） 本申请是一个名为“细胞内病原体和 先天免疫”我们要解决的中心问题是细胞内病原体是如何被细胞识别的。 宿主以及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，以促进其发病机制。在其 在以前的迭代中，该计划项目利用了新兴的全基因组技术， 表征巨噬细胞对多种细胞内细菌病原体的转录反应 并发现不同细胞内细菌病原体的感染导致STING的激活 和下游诱导的核心转录反应占主导地位的表达I型 干扰素调控基因为了更深入地探索宿主对细胞内病原体的反应， P01实验室已经开始研究对感染的转录后反应。在项目1，Portnoy 扩展了他对c-di-AMP和STING在单核细胞增生李斯特菌感染过程中的作用的研究， 免疫力他进一步提出研究如何L。单核细胞增生避免自噬以及这如何影响 发病机制和免疫诱导。最后，他建议对东道国和 在感染过程中细菌蛋白泛素化。在项目2中，考克斯研究了自噬体的形成 在含有结核分枝杆菌的吞噬体中被选择性激活。接下来，根据他的全球 泛素化研究将IRF 7鉴定为促进细胞内生长的转录因子， M.他将阐明IRF 7促进感染的机制。最后，他建议 鉴定M.肺结核感染。在项目3中， 万斯提出通过使用CRISPR/Cas9来解开STING激活的相反作用。 技术，以产生新的STING敲入小鼠，选择性地消除特定的STING依赖性 体内反应。下一步，他将确定嗜肺军团菌如何靶向中枢代谢 调节mTORC 1，以及它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中的分析技术，以确定新的转录后反应L。嗜肺菌核心 B，巴顿将提供总体方向和维持小鼠群体，繁殖小鼠，回交小鼠，并使用 CRISPR-Cas9产生新的基因组编辑的小鼠模型，以确定后基因的体内功能。 上面鉴定的转录反应。在核心C中，Krogan将应用新的基于质谱的， 生物信息学方法定量分析翻译后相互作用，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， 通过提供科学、组织和行政领导来促进协同作用， 通过在所有P01实验室和附属小组的月度会议上进行广泛的科学审查来完成。

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.