Host-pathogen Genetics Using Listeria and Drosophila

利用李斯特菌和果蝇进行宿主病原体遗传学

• 批准号: 7240516
• 负责人: Nancy Elizabeth Freitag
• 金额: $ 29.25万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7240516
• 关键词: Adult; Affect; Animals; Attenuated; Bacteria; Bacterial Proteins; Binding; Biological Models; Cell Line; Cells; Data; Defect; Drosophila genus; Drosophila melanogaster; Elements; Employee Strikes; Foundations; Genetic; Genetic Models; Genetic Screening; Goals; Growth; Host Defense; Human; Immune; Immune response; Infection; Infectious Agent; Insecta; Integration Host Factors; Knowledge; Larva; Listeria; Listeria monocytogenes; Listeriosis; Location; Mammalian Cell; Mammals; Microbe; Modeling; Mus; Mutation; Nature; Numbers; Organelles; Organism; Outcome; Pathogenesis; Pathway interactions; Predisposition; Proteins; Resistance; Resistance to infection; Role; Salmonella typhimurium; Screening procedure; Staging; System; Tissues; Vertebrates; Virulence; Work; bacterial genetics; base; cell type; design; fly; foodborne infection; microbial; mutant; pathogen; research study; response; tissue culture; tissue/cell culture

DESCRIPTION (provided by applicant): Listeria monocytogenes is a facultative intracellular bacterial pathogen that is an important agent of serious human food-borne infections. L. monocytogenes has served for decades as an outstanding model system for elucidating cellular and molecular interactions that take place during host infection. This proposal seeks to combine the power of bacterial genetics with the use of a genetically tractable model host system to isolate host mutants with altered resistance to L. monocytogenes infection. The fruit fly Drosophila melanogaster has been intensely studied as a model genetic system for decades, and it offers several advantages as a model host including the striking conservation of its innate immune recognition pathways with those of vertebrate animals. Studies have demonstrated the conservation of pathogenic mechanisms used by infectious agents within flies and vertebrates, and the feasibility of screening thousands of flies to isolate mutants with altered responses to infection provides a powerful means of identifying host factors that contribute to host survival. Recent work has demonstrated that D. melanogaster serves as a suitable host for L. monocytogenes infection, thus this proposal seeks to exploit the use of these genetically tractable organisms to functionally identify critical factors of both pathogen and host that contribute to the establishment of microbe infection. In Aim 1, experiments will examine the cellular course of L. monocytogenes infection within insect tissue culture cells, larvae, and adult flies. These studies will provide a foundation for the functional analysis of pathogen and host gene products identified for their potential roles in influencing the outcome of microbial infection. Aim 2 will functionally characterize L. monocytogenes mutants that are attenuated for virulence in flies. Bacterial gene products required for insect infection will be analyzed for potential roles in mammalian infection. Aim 3 will isolate and identify Drosophila mutants with altered susceptibility to Listeria infection. These studies will help identify the host factors that contribute to immune responses directed against intracellular pathogens. The ultimate goal of the experiments described will be the functional characterization of bacterial factors that support survival within the host, and the elucidation of host mechanisms that serve to counter bacterial survival strategies.

描述（由申请人提供）：单核细胞增生李斯特菌是一种兼性细胞内细菌病原体，是严重的人类食物传播感染的重要药物。单核细胞增生李斯特氏菌已成为一种出色的模型系统，用于阐明宿主感染期间发生的细胞和分子相互作用。该提案旨在将细菌遗传学的能力与使用遗传学模型的宿主系统相结合，以分离出对单核细胞增生李斯特氏菌感染的抗性改变的宿主突变体。数十年来，对果蝇果蝇的果蝇（Drosophila Melanogaster）进行了深入研究，它作为模型宿主提供了多种优势，包括对其先天免疫识别途径与脊椎动物的固有免疫识别途径。研究表明，蝇和脊椎动物内感染剂使用的致病机制的保存，以及筛查数千只苍蝇以分离出对感染反应改变的突变体的可行性，提供了一种有力的宿主因素，以识别有助于宿主存活的宿主因素。最近的工作表明，黑色素芽孢杆菌是单核细胞增生李斯特氏菌感染的合适宿主，因此该提案旨在利用这些遗传障碍生物的使用，以在功能上识别病原体和宿主的关键因素，从而有助于建立微生物感染。 在AIM 1中，实验将检查昆虫组织培养细胞，幼虫和成年果蝇中单核细胞增生乳杆菌感染的细胞病程。这些研究将为病原体和宿主基因产物的功能分析提供基础，这些基因在影响微生物感染结果中的潜在作用。 AIM 2在功能上将表征单核细胞增生李斯特氏菌突变体，这些突变体因蝇中的毒力而衰减。将分析昆虫感染所需的细菌基因产品，以在哺乳动物感染中的潜在作用。 AIM 3将分离并鉴定果蝇突变体对李斯特菌感染的敏感性改变。这些研究将有助于确定有助于针对细胞内病原体的免疫反应的宿主因素。所描述的实验的最终目标是支持宿主内生存的细菌因子的功能表征，以及阐明用于抵抗细菌生存策略的宿主机制。