Project 1: Listeria metabolites and innate immunity

项目1：李斯特菌代谢物与先天免疫

• 批准号: 10190578
• 负责人: DANIEL A PORTNOY
• 金额: $ 49.85万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-30 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10190578
• 关键词: Address; Alleles; Anabolism; Antigens; Bacillus subtilis; Bacteria; Bacterial Infections; Binding; Biological Models; Brain; Cells; Cellular biology; Communicable Diseases; Communities; Data; Disease; Electron Transport; Enterocolitis; Flavins; Funding; Genes; Goals; Human; Immune response; Immune system; Immunity; Infection; Innate Immune Response; Innate Immune System; Interferon Type I; Interferons; International; Learning; Legionella pneumophila; Listeria; Listeria monocytogenes; Listeriosis; Longitudinal Studies; Mediating; Metabolism; Microbiology; Modeling; Mucous Membrane; Mus; Mycobacterium tuberculosis; Natural Immunity; Nucleotides; Nutritional Requirements; Oral; Pathogenesis; Pathway interactions; Patient observation; Periodicity; Placenta; Play; Prevention; Primates; Regulation; Riboflavin; Role; Shapes; Signal Transduction; Signaling Molecule; System; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Tuberculosis; Tuberculosis Vaccines; Vaccines; adaptive immunity; bacterial metabolism; base; efflux pump; extracellular; foodborne pathogen; global health; innate immune pathways; interest; isoprenoid; mevalonate; microorganism; mutant; novel; oral infection; pathogen; placental infection; prevent; response; tissue culture; tuberculosis immunity; vaccine development; γδ T cells

Project Summary/Abstract (Project 1, Portnoy) This proposal is Project 1 within a P01 renewal, entitled “The intersection of innate and adaptive immunity to intracellular pathogens.” A central problem that we address is how intracellular pathogens are recognized by the host innate immune system and how multiple signals are integrated to induce an appropriate response, and conversely, how pathogens avoid and/or manipulate host responses to promote their pathogenesis. In Project 1, we have chosen to approach this problem by continuing a detailed analysis of Listeria monocytogenes, a facultative intracellular, food-borne pathogen that has been studied for decades as a model system with which to dissect basic aspects of infection & immunity. Previously, we discovered that L. monocytogenes secretes c-di-AMP which binds to and activates host STING leading to a type I interferon response. In Aim 1, we exploit a newly developed oral model of listeriosis where we find that bacterially secreted c-di-AMP induces a STING-dependent host protective response. We propose to analyze how STING provides protection and test the hypothesis that STING also promotes dissemination to the placenta and brain. In Aims 2 and 3, we begin to explore the idea that c-di-AMP might be just one example of the principle that small bacterial metabolites play important roles in shaping host immunity. L. monocytogenes is a riboflavin requiring microorganism, and we explore the hypothesis that that this nutritional requirement allows the bacteria to avoid activation of mucosal-associated invariant T-cells (MAIT cells), which are innate T-cells stimulated by a modified intermediate of riboflavin biosynthesis. In preliminary data, we introduced five Bacillus subtilis genes (ribDEAHT) into L. monocytogenes converting it into a riboflavin-synthesizing bacterium and are now poised to test our hypothesis and learn about the role of MAIT cells during bacterial infection and immunity. We also propose to explore the activation of the dominant gamma delta T-cell in humans (Vg9Vd2 T-cells). These cells are activated by an intermediate of bacterial isoprenoid biosynthesis (HMBPP). Our ultimate goal is to determine the importance of this innate immune pathway during infection of primates (human and non-human). Here we propose to construct bacterial mutants that either fail to make HMBPP or make more of it to examine how bacterial HMBPP is detected during infection in tissue culture models. We further propose to collaborate with the Cox lab (Project 2) to explore the importance of HMBPP during M. tuberculosis infection and immunity.

项目摘要/摘要（项目 1，波特诺伊） 该提案是 P01 更新中的项目 1，题为“先天免疫和适应性免疫的交叉点” 细胞内病原体。”我们要解决的一个中心问题是如何识别细胞内病原体 宿主先天免疫系统以及如何整合多个信号以诱导适当的免疫反应 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。在项目 1 中，我们选择通过继续详细分析来解决这个问题 单核细胞增生李斯特氏菌，一种兼性细胞内食源性病原体，已被研究数十年 作为剖析感染和免疫基本方面的模型系统。此前，我们发现 单增李斯特菌分泌 c-di-AMP，它结合并激活宿主 STING，导致 I 型 干扰素反应。在目标 1 中，我们利用了一种新开发的李斯特菌病口腔模型，我们发现 细菌分泌的 c-di-AMP 诱导 STING 依赖性宿主保护反应。我们建议 分析 STING 如何提供保护并检验 STING 还促进传播的假设 胎盘和大脑。在目标 2 和 3 中，我们开始探索 c-di-AMP 可能只是其中之一的想法 小细菌代谢物在塑造宿主免疫力方面发挥重要作用的原理示例。 L。 单核细胞增生李斯特菌是一种需要核黄素的微生物，我们探索了这样的假设： 营养需求使细菌能够避免激活粘膜相关的不变 T 细胞 (MAIT 细胞），它们是由核黄素生物合成的修饰中间体刺激的先天 T 细胞。在 初步数据，我们将五个枯草芽孢杆菌基因（ribDEAHT）引入单核细胞增生李斯特菌中，将其转化 进入核黄素合成细菌，现在准备测试我们的假设并了解其作用 MAIT细胞在细菌感染和免疫过程中的作用。我们还建议探索激活 人类中的显性 γ δ T 细胞（Vg9Vd2 T 细胞）。这些细胞被中间体激活 细菌类异戊二烯生物合成（HMBPP）。我们的最终目标是确定这种与生俱来的重要性 灵长类动物（人类和非人类）感染期间的免疫途径。这里我们建议构建 无法产生 HMBPP 或产生更多 HMBPP 的细菌突变体，以检查细菌 HMBPP 的性质 在组织培养模型感染期间检测到。我们进一步提议与考克斯实验室合作 （项目2）探讨HMBPP在结核分枝杆菌感染和免疫过程中的重要性。