Intracellular pathogens and innate immunity

项目总监/主要调查员（最后，（最后，第一，中间）：Portnoy，Daniel 2 P01 AI 063302 -06项目总监/主要调查员第一，中间）：Portnoy，丹尼尔A。A. 2 P01 A1063302-06 摘要 更新P01本申请是作为P01的竞争性更新提交的，P01最初是为了响应N PAID作为NIAID生物防御和新兴传染病公告（PA）的一部分的计划公告（PA）而提交的，NIAID是生物防御和研究计划的一部分。拟议的研究是集中在五个不同的兼性研究计划的相互作用。拟议的研究重点是五种不同的兼性细胞内微生物病原体与巨噬细胞的相互作用，这是传染病研究的一个中心问题，也是传染病研究的一个中心问题，对全球健康，生物防御和新出现的感染具有重要影响。拟议的研究依赖于全球健康，生物防御和新出现的感染的重要分支。拟议的研究在很大程度上依赖于使用细菌和宿主突变体来检查巨噬细胞胞质监视途径，以检测细胞内病原体的先天免疫的巨噬细胞胞质监视途径。这个P01的中心假设是微生物的先天免疫检测病原体。该P01的中心假设是，通过辅助分泌系统分泌或释放的微生物分子被宿主特异性识别，通过辅助分泌系统分泌释放的微生物分子被宿主受体特异性识别，导致IRF 3胞质受体活化。这些受体的刺激导致1 RF 3和随后的受体的激活。IFNb和共调节基因的表达。该模型提供了宿主细胞表达IFNb和共调节基因的可能机制。该模型提供了一种可能的机制，通过该机制宿主细胞能够检测由活的复制微生物分泌的微生物产物。具体目标是：一。由活的复制微生物分泌的具体目标是：一。通过检查巨噬细胞对结核分枝杆菌、荚膜组织胞浆菌、Francise/la tularensis和单核细胞增生李斯特菌的转录反应，通过检查巨噬细胞对分枝杆菌先天免疫识别的转录反应，鉴定和表征有助于刺激先天免疫识别的宿主胞质途径的分子决定簇。结核病、荚膜组织胞浆菌、土拉热弗朗西斯菌和单核细胞增生李斯特菌的鉴定。将使用正向遗传筛选和检测IFN-b的生物测定法（正向和生物测定IFN-b，一种对胞质途径具有特异性的细胞因子）来完成这些决定簇的鉴定。二.鉴定激活细胞溶质途径特异性细胞因子的宿主途径和细菌配体。二.识别激活胞质应答的宿主途径和菌配体。二.描述先天性免疫识别的胞质途径在胞质III中的作用。在动物感染模型中的免疫识别途径。目标II和III将由核心C（动物核心）支持。四.建立常见感染动物模型。目的II和III（动物核心）。四.建立共同和独特的宿主转录谱，建立对微生物病原体的签名响应。利用对微生物病原体的反应。利用转录特征识别宿主反应的共同和不同途径和机制。目的共同和途径和机制宿主反应。目标IV将得到核心B（转录谱核心）的支持。虽然这个提议没有直接IV将支持核心B（转录谱核心）。虽然本提案不直接涉及BSL 2和BSL 3病原体，因此，每个PI涉及选定的试剂，但它确实涉及BSL 2和BSL 3病原体，因此，每个P1都描述了其确保生物安全性的承诺。这些目标代表着从最初的生物安全赠款范围缩小。这些目标代表了从最初的赠款提交范围缩小，以专注于可以在两年内完成的关键实验。请参阅提交的重点是可以在两年内完成的关键实验。请参阅修订版

Program Director/Principallnvestigator (Last, (Last, First, Middle): Portnoy, Daniel2 P01 AI063302-06 Program Director/Principal InVestigator First, Middle): Portnoy, Daniel A. A. 2 P01 A1063302-06 ABSTRACT renewal P01 This application was submitted as a competitive renewal of a P01 originally submitted in response to a submitted in response to a Program Announcement (PA) from N PAIDas part of the Biodefense and Emerging Infectious Disease Disease Announcement (PA) from NIAID as part of the Biodefense and Research Program. The proposed research is focused on the interaction of five diverse facultative Research Program. The proposed research is focused on the interaction of five diverse facultative intracellular microbial pathogens with macrophages, a central problem of infectious disease research, with a central problem of infectious disease research, with important ramifications for global health, biodefense and emerging infections. The proposed research relies important ramifications for global health, biodefense and emerging infections. The proposed research relies heavily on the use of bacterial and host mutants to examine a macrophage cytosolic surveillance pathway of heavily on the use bacterial and host mutants to examine a macrophage cytosolic surveillance pathway of innate immunity that detects intracellular pathogens. A central hypothesis of this P01 is that microbial innate immunity that detects pathogens. A central hypothesis of this P01 is that microbial molecules, secreted or released through auxiliary secretion systems, are specifically recognized by host secreted released through auxiliary secretion systems, are specifically recognized by host receptors leads to the activation of IRF3 cytosolic receptors. Stimulation of these receptors leads to the activation of 1RF3 and the subsequent receptors. expression of IFNb and co-regulated genes. This model provides a possible mechanism by which host cells expression lFNb and co-regulated genes. This model provides a possible mechanism by which host cells are able to detect microbial products secreted by live, replicating microorganisms. The Specific Aims are: I. secreted by live, replicating microorganisms. The Specific Aims are: I. Identify and characterize molecular determinants that contribute to stimulating the host cytosolic pathways of Identify and characterize determinants that contribute to stimulating the host cytosolic pathways of innate immune recognition by examining macrophage transcriptional responses to Mycobacterium innate immune recognition by examining macrophage transcriptional responses to Mycobacterium tuberculosis, Histoplasma capsulatum, Francise/la tularensis, and Listeria monocytogenes. Identification of tuberculosis, Histoplasma capsulatum, Francisella tularensis, and Listeria monocytogenes. Identification of these determinants will be accomplished using forward genetic screens and a bioassay that detects IFN-b, a forward and bioassay IFN-b, a cytokine specific to the cytosolic pathway. II. Identify host pathways and bacterial ligand(s) that activate the cytokine specific to the cytosolic pathway. II . Identify host pathways and bacterialligand(s) that activate the cytosolic response. II I. Characterize the role(s) of the cytosolic pathways of innate immune recognition in role(s) cytosolic Ill. pathways immune recognition in animal models of infection. Aims II and III will be supported by Core C (animal core). IV. Establish common animal models infection. Aims II and Ill by (animal core). IV. Establish common and unique host transcription profiles that establish signature responses to microbial pathogens. Utilize responses to microbial pathogens. Utilize transcriptional signatures to identify common and distinct pathways and mechanisms of host response. Aim common and pathways and mechanisms host response. Aim IV will be supported by Core B (transcriptional profiling core). Although this proposal does not directly IV will be supported by Core B (transcriptional profiling core). Although this proposal does not directly it does involve BSL2 and BSL3 pathogens and accordingly, each PI involve select agents, it does involve BSL2 and BSL3 pathogens and accordingly, each P1 has described their commitment to ensure biosafety. The aims represent a reduction in scope from the original grant biosafety. The aims represent a reduction in scope from the original grant submission in order to focus on critical experiments that can be accomplished in two years. Please see the submission focus on critical experiments that can be accomplished in two years. Please see the Revised