Francisella tularensis intracellularly-induced outer membrane proteins

土拉弗朗西斯菌细胞内诱导的外膜蛋白

基本信息

批准号：
8115148
负责人：
Jason F Huntley
金额：
$ 10.8万
依托单位：
UNIVERSITY OF TOLEDO HEALTH SCI CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8115148
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 Reverse Transcriptase Polymerase Chain Reaction 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

项目摘要

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）：抽象的 =================== 注意：此摘要是从应用程序中提取的，尚未由SRA证明。当应用程序扫描过程存在问题时，提取的文本可能不正确或不完整。 =================== Francisella tuarlarensis是tularemia的病因学药物，是一种革兰氏阴性的细胞内球杆菌，擅长于包括人类在内的许多哺乳动物物种中诱导急性致命疾病。 F. tularensis被广泛认可为最危险的细菌病原体之一，因为其感染剂量低（<10个生物），易感性，多种感染途径以及诱导严重的发病率和死亡率的能力。关于F. tularensis用来侵袭宿主细胞并随后引起疾病的分子知之甚少，但是外膜蛋白（OMP）可能在此过程中有助于。实际上，在许多其他细菌病原体中，OMP已被证明是毒力因子，因此，是理想的疫苗靶标。在博士后研究期间，我开发了一种提取和净化tularensis Omps的方法，这导致识别近30个新的OMP。在随后的研究中，我证明了一种基于OMP的疫苗保护小鼠免受毒性A型F. tularensis肺挑战的影响。尽管这些结果令人信服，并且在Tularemia研究领域已经大大推进了这些结果，但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，但是合理的假设是flularensis在体内显着改变其OMP的特征以促进宿主细胞侵袭和疾病。因此，这项研究建议将通过以下方式扩展我以前的发现：（1）使用RRT-PCR检查哺乳动物宿主中已知的OMP是上调还是下调；（2）发现在巨噬细胞中专门表达的新OMP；（3）通过产生缺陷的突变体来识别新的毒力因子，并评估这些突变体在小鼠肺部感染模型中的感染性；（4）表征抗体介导的和细胞介导的免疫反应，以赋予抗tularensis肺挑战的OPM。这些项目旨在回答有关F. tularensis毒力以及保护性免疫相关的基本问题。此外，该提案在开发安全，有效的疫苗以防止F. tularensis感染和疾病方面有实际应用。弗朗西斯菌（Francisella toarlensis）不仅重要，这不仅是因为其将其分类为选择剂，而且还因为其迅速的细胞内发病机理。关于F. tularensis的表面分子知之甚少，但是这些信息可以为开发安全，有效的疫苗针对Tularemia提供重要信息。该提案中概述的项目旨在识别涉及感染的细菌分子并表征保护性免疫反应。