YopM and protective innate defenses against plague

YopM 和针对鼠疫的保护性先天防御

• 批准号: 8017377
• 负责人: SUSAN C STRALEY
• 金额: $ 47.13万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-16 至 2013-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8017377
• 关键词: Biology; Bubonic Plague; CD8B1 gene; Cells; Cytokine Signaling; Cytotoxic T-Lymphocytes; Data; Dendritic Cells; Down-Regulation; Effector Cell; Exposure to; Foundations; Human; Immune; Immune response; Infection; Interleukin-1; Interleukin-12; Interleukin-15; Interleukin-18; Knockout Mice; Lung; Maps; Mediating; Messenger RNA; Mus; Natural Killer Cells; Parents; Pathway interactions; Plague; Pneumonic Plague; Population; Predictive Value; Reporting; Resistance; Signal Pathway; Signal Transduction; Spleen; T-Lymphocyte; Testing; Therapeutic Intervention; Time; Toxin; Virulence; Virulent; Yersinia pestis; cDNA Arrays; chemokine; cytokine; insight; mRNA Expression; macrophage; microbial host; mouse model; mutant; neutrophil; pathogen; prevent; research study

DESCRIPTION (provided by applicant): Yersinia pestis, the causative agent of plague, delivers a set of 6 Yop toxins that act within host cells to negate and modulate signaling pathways that orchestrate host innate defenses that are crucial for developing an effective immune response. This proposal focuses on the mechanism of YopM. We have found that YopM specifically causes a global loss of natural killer cells, and those NK cells remaining do not contain detectable mRNA for IFNg. Concomitantly, YopM causes down-regulation in macrophages of net mRNA expression for key proinflammatory cytokines. In this project, we will test the hypothesis that YopM acts directly on cells such as macrophages and dendritic cells to prevent expression of cytokines such as IL- 15 and IL-12 that synergize to activate effector cells such as macrophages, polymorphonuclear neutrophils, natural killer cells, and cytolytic T cells for controlling numbers of Y. pestis. In Aim 1, we will determine whether YopM's action is mediated by hypothesized key cells by comparing the infection dynamics for virulent and YopM- Y. pestis (viable numbers, dissemination as a function of time) and host response (cell populations and cytokines) in primary pneumonic plague and bubonic plague in mice ablated for the cells genetically or by manipulation. In Aim 2, we will test our hypothesis that the Th1 cytokine and chemokine pathway mediates different host responses to YopM+ and YopM- Y. pestis between d 1 and d 3 to 4 p.i. in bubonic and pneumonic plague by infecting knockout mice or mice ablated for hypothesized key cytokines and analyzing the resulting infection dynamics and host response. In Aim 3, we will map the signaling circuitry of YopM's pathogenic effects by comparing transcriptional profiles (cDNA microarrays) and analyses of intracellular cytokines in purified macrophages from mice with pneumonic or bubonic plague due to virulent and YopM- Y. pestis. The data will reveal the pathogenic mechanism of YopM by identifying cells, cytokines, and signaling pathways modulated by YopM during plague. The findings will provide a completely new understanding of the biology of plague from both microbial and host perspectives as well as provide a framework for rational enhancement of resistance to plague in people threatened with exposure to plague. The data will reveal immune deviation points during plague and could identify potential therapeutic intervention targets. The findings also will provide a better understanding of mammalian innate defenses and insight into evasive mechanisms of pathogens and will facilitate the defeat of important human pathogens that are not select agents.

描述（由申请人提供）：鼠疫耶尔森氏菌是鼠疫的病原体，它提供一组 6 种 Yop 毒素，这些毒素在宿主细胞内起作用，以否定和调节信号通路，从而协调宿主先天防御，这对于形成有效的免疫反应至关重要。本提案重点关注YopM的机制。我们发现 YopM 特异性地导致自然杀伤细胞的整体丧失，而剩下的 NK 细胞不含有可检测到的 IFNg mRNA。与此同时，YopM 导致巨噬细胞中关键促炎细胞因子的净 mRNA 表达下调。在这个项目中，我们将测试这样的假设：YopM 直接作用于巨噬细胞和树突状细胞等细胞，以阻止 IL-15 和 IL-12 等细胞因子的表达，这些细胞因子协同激活巨噬细胞、多形核中性粒细胞、自然杀伤细胞等效应细胞细胞和溶细胞 T 细胞，用于控制鼠疫耶尔森氏菌的数量。在目标 1 中，我们将通过比较强毒力和 YopM-鼠疫菌的感染动态（活菌数量、随时间变化的传播）和宿主反应（细胞群和细胞因子）来确定 YopM 的作用是否由假设的关键细胞介导。小鼠的原发性肺鼠疫和腺鼠疫通过基因或操作去除细胞。在目标 2 中，我们将检验我们的假设，即 Th1 细胞因子和趋化因子途径在感染后第 1 天和第 3 至 4 天之间介导对 YopM+ 和 YopM- 鼠疫耶尔森氏菌的不同宿主反应。在腺鼠疫和肺鼠疫中，通过感染基因敲除小鼠或消除假设的关键细胞因子的小鼠并分析由此产生的感染动态和宿主反应。在目标 3 中，我们将通过比较转录谱（cDNA 微阵列）和分析纯化的巨噬细胞中的细胞内细胞因子，绘制 YopM 致病效应的信号通路，这些巨噬细胞来自患有由强毒力和 YopM-Y. 鼠疫引起的肺鼠疫或黑死病的小鼠。这些数据将通过识别鼠疫期间 YopM 调节的细胞、细胞因子和信号通路来揭示 YopM 的致病机制。这些发现将从微生物和宿主的角度提供对鼠疫生物学的全新理解，并为合理增强受到鼠疫威胁的人们对鼠疫的抵抗力提供​​框架。这些数据将揭示鼠疫期间的免疫偏差点，并可以确定潜在的治疗干预目标。这些发现还将提供对哺乳动物先天防御的更好理解，并深入了解病原体的逃避机制，并将有助于击败非选择性病原体的重要人类病原体。

项目成果

YopM and plague.

YopM 和瘟疫。

• DOI: 10.1007/978-1-4614-3561-7_31
• 发表时间: 2012
• 期刊: Advances in experimental medicine and biology
• 作者: Straley,SusanC

Toward a molecular pathogenic pathway for Yersinia pestis YopM.

• DOI: 10.3389/fcimb.2012.00155
• 发表时间: 2012
• 期刊: Frontiers in cellular and infection microbiology
• 影响因子: 5.7
• 作者: Uittenbogaard AM;Chelvarajan RL;Myers-Morales T;Gorman AA;Brickey WJ;Ye Z;Kaplan AM;Cohen DA;Ting JP;Straley SC
• 通讯作者: Straley SC