Modulation of Salmonella Infection by Focal Adhesion Kinase

粘着斑激酶对沙门氏菌感染的调节

• 批准号: 7679219
• 负责人: Katherine A. Owen
• 金额: $ 4.72万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7679219
• 关键词: Acute; Affect; Animals; Anti-Inflammatory Agents; Anti-inflammatory; Antibiotic Resistance; Attenuated; Bacteria; Biological Models; Bone Marrow; Cells; Cessation of life; Complex; Cultured Cells; Data; Dendritic Cells; Disease; Distal; Epithelial; Epithelial Cells; Equilibrium; Focal Adhesion Kinase 1; Focal Adhesions; Gastroenteritis; Genes; Goals; Health; Host Defense; Human; Immune response; Infection; Infection Control; Infiltration; Inflammation; Inflammatory; Inflammatory disease of the intestine; Integrins; Interleukin-1; Interleukin-6; Interleukin-8; Intestines; Investigation; Knockout Mice; Lamina Propria; Liver; Mediating; Membrane; Messenger RNA; Modeling; Mus; Myelogenous; Organism; Pathogenesis; Pathology; Pattern; Phagocytes; Phase; Play; Population; Predisposition; Production; Proteins; Receptor Protein-Tyrosine Kinases; Regulation; Research; Role; Salmonella; Salmonella infections; Salmonella typhimurium; Septicemia; Severities; Signal Pathway; Signal Transduction; Signaling Molecule; Site; Small Intestines; Spleen; Staging; Techniques; Tissues; Tumor Necrosis Factor-alpha; Tumor Necrosis Factors; Typhoid Fever; cell motility; chemokine; cytokine; directional cell; enteric pathogen; human disease; ileum; in vivo; intestinal epithelium; knockout animal; macrophage; migration; monocyte; mouse model; neutrophil; pathogen; research study; resistant strain; villin

DESCRIPTION (provided by applicant): Bacteria of the genus Salmonella are a group of gram-negative pathogens that cause a wide range of human diseases from localized gastroenteritis to typhoid fever and septicemia. Although significant progress has been made in understanding the mechanisms of Salmonella pathogenesis, more than 12 million cases of typhoid fever occur annually, and emerging antibiotic resistant strains pose a significant threat to human health. The overall goal of the proposed research is to understand how the non-receptor tyrosine kinase Focal Adhesion Kinase (FAK) functions in host defense against the enteric pathogen S. typhimurium. The experiments outlined herein provide an integrated approach to determine how this molecule affects Salmonella disease pathology through its impact on the innate immune response and the integrity of the intestinal epithelial barrier. In Specific Aim 1, we will determine how FAK expression by macrophages modulates the cellular immune response during Salmonella infection in vivo. We will accomplish this by examining bacterial colonization levels and inflammation in tissues derived from wildtype and myeloid-specific conditional FAK knockout mice. We will also examine the phagocytic capacity of FAK-deficient macrophages using advanced flow cytometric techniques. The studies described in this aim will elucidate how differences in the recruitment and/or function of phagocytes occurring in the absence of FAK impact the complex balance between host survival and death. While the presence of cytokines helps initiate and regulate the immune response to Salmonella, the appropriate balance between pro- and anti-inflammatory cytokines is essential to control infection and avoid damage to the host. In Specific Aim 2, we will investigate the FAK-mediated signaling pathways involved in macrophage-mediated cytokine production during Salmonella infection. To do this, we will first examine the ability of FAK-deficient macrophages to produce pro- and anti-inflammatory cytokines followed by an investigation into the specific signaling molecules involved in cytokine production. Finally, in Specific Aim 3, we will determine how FAK expression in the intestinal epithelium impacts host susceptibility to Salmonella infection. Using mice in which FAK is conditionally deleted from the intestinal epithelium, we will assess how the loss of this molecule from the epithelial barrier affects bacterial colonization levels in the small intestine and dissemination to distal tissues.

描述（由申请人提供）：沙门氏菌属细菌是一组革兰氏阴性病原体，可引起多种人类疾病，从局部胃肠炎到伤寒和败血症。尽管在了解沙门氏菌发病机制方面已取得重大进展，但每年仍有超过 1200 万例伤寒病例发生，新出现的抗生素耐药菌株对人类健康构成重大威胁。拟议研究的总体目标是了解非受体酪氨酸激酶粘着斑激酶（FAK）如何在宿主防御肠道病原体鼠伤寒沙门氏菌中发挥作用。本文概述的实验提供了一种综合方法来确定该分子如何通过其对先天免疫反应和肠上皮屏障完整性的影响来影响沙门氏菌疾病病理学。在具体目标 1 中，我们将确定巨噬细胞的 FAK 表达如何在体内沙门氏菌感染期间调节细胞免疫反应。我们将通过检查野生型和骨髓特异性条件 FAK 敲除小鼠组织中的细菌定植水平和炎症来实现这一目标。我们还将使用先进的流式细胞术技术检查 FAK 缺陷型巨噬细胞的吞噬能力。该目标中描述的研究将阐明在缺乏 FAK 的情况下吞噬细胞的招募和/或功能的差异如何影响宿主生存和死亡之间的复杂平衡。虽然细胞因子的存在有助于启动和调节对沙门氏菌的免疫反应，但促炎细胞因子和抗炎细胞因子之间的适当平衡对于控制感染和避免对宿主造成损害至关重要。在具体目标 2 中，我们将研究沙门氏菌感染期间参与巨噬细胞介导的细胞因子产生的 FAK 介导的信号通路。为此，我们将首先检查 FAK 缺陷型巨噬细胞产生促炎和抗炎细胞因子的能力，然后研究参与细胞因子产生的特定信号分子。最后，在具体目标 3 中，我们将确定肠上皮中的 FAK 表达如何影响宿主对沙门氏菌感染的易感性。使用从肠上皮有条件删除 FAK 的小鼠，我们将评估上皮屏障中该分子的丢失如何影响小肠中的细菌定植水平以及向远端组织的传播。