Francisella tularensis intracellularly-induced outer membrane proteins

土拉弗朗西斯菌细胞内诱导的外膜蛋白

• 批准号: 7882067
• 负责人: Jason F Huntley
• 金额: $ 15.77万
• 依托单位: UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7882067
• 关键词: Acute; Antibiotics; Antibodies; B-Lymphocytes; Bacteria; Binding; Categories; Cells; Centers for Disease Control and Prevention (U.S.); Classification; Conditioned Culture Media; Detection; Development; Disease; Dose; Engineering; Francisella tularensis; Generations; Human; Immune; Immune response; Immunity; Immunization; In Vitro; Individual; Infection; Invaded; Lead; Lung; Mass Spectrum Analysis; Mediating; Membrane; Membrane Proteins; Methods; Modeling; Molecular Profiling; Morbidity - disease rate; Mus; Organism; Pathogenesis; Process; Production; Proteins; Proteomics; Recombinants; Regimen; Reporting; Research; Research Proposals; Route; Scanning; Scientist; Splenocyte; Surface; System; Testing; Text; Tularemia; Two-Dimensional Gel Electrophoresis; Vaccines; Virulence; Virulence Factors; Virulent; abstracting; base; cell mediated immune response; cytokine; design; in vivo; macrophage; mortality; mutant; pathogen; practical application; prevent; protein expression; protein profiling; resistant strain; response

DESCRIPTION (provided by applicant): Abstract ================== NOTICE: THIS ABSTRACT WAS EXTRACTED FROM APPLICATION AND HAS NOT BEEN PROOFED BY AN SRA.WHEN THERE ARE PROBLEMS WITH THE APPLICATION SCANNING PROCESS, THE EXTRACTED TEXT MAY BE INCORRECT OR INCOMPLETE. ================== Francisella tularensis, the etiological agent of tularemia, is a Gram-negative intracellular coccobacillus that is adept at inducing acute, fatal disease in a number of mammalian species, including humans. F. tularensis is well-recognized as one of the most dangerous bacterial pathogens known because of its low infectious dose (< 10 organisms), ease of aerosolization, multiple routes of infection, and capacity for inducing severe morbidity and mortality. Little is known about the molecules that F. tularensis uses to invade host cells and subsequently cause disease, but outer membrane proteins (OMPs) are likely instrumental in this process. Indeed, OMPs have been shown to be virulence factors in many other bacterial pathogens and, for this reason, are ideal vaccine targets. During my postdoctoral research, I developed a method to extract and purify F. tularensis OMPs, which has led to the identification of nearly 30 new OMPs. In subsequent studies, I demonstrated that an OMP-based vaccine protected mice against virulent Type A F. tularensis pulmonary challenge. While these results are compelling and have greatly advanced the tularemia research field, the bacteria used for these studies were grown under rich media conditions. To date, little has been reported about OMP expression during host infection, yet it is reasonable to hypothesize that F. tularensis dramatically alters its OMP profile in vivo to facilitate host cell invasion and disease. As such, this research proposal will extend my previous findings by: (1) Using rRT-PCR to examine whether known OMPs are up- or down-regulated in the mammalian host; (2) Discovering new OMPs that are exclusively expressed in macrophages; (3) Identifying new virulence factors through the generation of OMP-deficient mutants and assessing the infectivity of these mutants in a mouse pulmonary infection model; and (4) Characterizing both the antibody-mediated and cell- mediated immune responses against OMPs that confer protection against F. tularensis pulmonary challenge. These project subaims are designed to answer fundamental questions about F. tularensis virulence as well as the correlates of protective immunity. Additionally, this proposal has practical applications for the development of safe, efficacious vaccines to prevent F. tularensis infection and disease. Francisella tularensis is important not only because of its classification as a Category A Select Agent, but because of its rapid intracellular pathogenesis. Little is known about the surface molecules of F. tularensis, yet such information could provide vital information for the development of a safe, efficacious vaccine against tularemia. The projects outlined in this proposal seek to identify bacterial molecules involved in infection and characterize protective immune responses.

描述（由申请人提供）： 摘要 ================== 注意事项：本摘要摘自应用程序，未经SRA验证。当应用程序扫描过程出现问题时，提取的文本可能不正确或不完整。 ================== 土拉热弗朗西丝氏菌（Francisella tularensis）是一种革兰氏阴性细胞内球杆菌，其擅长在包括人类在内的许多哺乳动物物种中诱导急性、致命性疾病。F.土拉热病被公认为已知的最危险的细菌病原体之一，因为其感染剂量低（< 10个生物体）、易于雾化、多种感染途径和诱导严重发病率和死亡率的能力。我们对F.土拉菌用于侵入宿主细胞并随后引起疾病，但外膜蛋白（OMP）可能在此过程中起作用。事实上，OMP已被证明是许多其他细菌病原体中的毒力因子，因此是理想的疫苗靶标。在我的博士后研究期间，我开发了一种提取和纯化F。土拉热OMP，这导致了近30个新的OMP的鉴定。在随后的研究中，我证明了一种基于OMP的疫苗可以保护小鼠免受A型F病毒的攻击。土拉热肺激发试验。虽然这些结果是令人信服的，并大大推进了兔热病研究领域，用于这些研究的细菌是在丰富的培养基条件下生长的。迄今为止，关于宿主感染过程中OMP表达的报道很少，但有理由假设F。土拉热菌在体内显著改变其OMP谱以促进宿主细胞侵袭和疾病。因此，这项研究计划将通过以下方式扩展我以前的研究结果：（1）使用rRT-PCR检测已知的OMP在哺乳动物宿主中是否上调或下调;（2）发现仅在巨噬细胞中表达的新OMP;（3）通过产生OMP缺陷突变体来鉴定新的毒力因子，并在小鼠肺部感染模型中评估这些突变体的感染性;和（4）表征针对赋予针对F的保护的OMP的抗体介导的和细胞介导的免疫应答。土拉热肺部挑战。这些项目子目标旨在回答有关F.土拉菌毒力以及保护性免疫的相关性。此外，这一建议对开发安全、有效的疫苗以预防F。土拉菌感染和疾病。土拉热弗朗西丝氏菌之所以重要，不仅是因为其被归类为A类选择性病原体，而且还因为其快速的细胞内发病机制。对F的表面分子知之甚少。然而，这些信息可以为开发一种安全、有效的兔热病疫苗提供重要信息。本提案中概述的项目旨在确定感染中涉及的细菌分子并描述保护性免疫反应。