Identification of secreted proteins important for tulremia pathogenesis

描述(申请人提供)：革兰氏阴性杆菌图拉氏亚种。图拉氏菌是图拉热症的病原体，被认为是一种潜在危险的生物武器。这是由于有机体通过气雾剂途径具有极强的传染性，以及图拉热症的严重性。这种细菌的挑剔性质，要求使用生物安全三级(BSL3)遏制来处理它，以及缺乏遗传工具，这些都阻碍了对这种有机体的研究。因此，与许多其他细菌病原体相比，人们对图拉夫酵母亚种知之甚少。图拉尔人的生理学、遗传学和发病机制。以前的研究使用的是图拉氏丝孢子虫亚种的减毒活疫苗株。Holarctica，或罕见致病的F·tularensis亚种。诺维西达。这项建议的目的是鉴定在图拉氏华支杆菌亚种致病过程中重要的分泌蛋白。图拉尔人。我们将使用最近开发的遗传工具来识别和干扰编码分泌蛋白质的基因，直接目标是识别小鼠巨噬细胞细胞内生存和复制所需的蛋白质。我们预计这项工作将确定几个感兴趣的蛋白质，并将使我们能够为进一步工作的基础制定假设。这项申请中提出的工作将有助于NIAID A类CDC制剂(如土拉方杆菌)的生物防御研究议程的研究目标。这些近期的目标包括：1)开发遗传系统，将发病机制和毒力的差异联系起来；2)增加图拉热症研究的专业知识的多样性。 这项研究将为图拉氏菌生物学的研究提供更多的遗传学方法，并确定图拉热病发病机制的重要因素。这项工作还可能有助于更好的诊断、疫苗和药物开发。由于图拉氏丝虫是一种选择性试剂，可以被用作生物武器，这项研究可能会对未来的公共卫生产生重大影响。

DESCRIPTION (provided by applicant): The Gram-negative bacterium Francisella tularensis subsp. tularensis, the causative agent of tularemia, is considered a potentially dangerous biological weapon. This is due to the organism's extreme infectivity via the aerosol route and the severity of tularemia. The fastidious nature of the bacterium, the requirement that it be handled using biosafety level three (BSL3) containment, and the paucity of genetic tools have hindered research on this organism. Thus, compared to many other bacterial pathogens, little is known about F. tularensis subsp. tularensis physiology, genetics, and pathogenesis. Previous studies have used the attenuated live vaccine strain (LVS) derived from F. tularensis subsp. holarctica, or the rarely pathogenic F. tularensis subsp. novicida. The goal of this proposal is to identify secreted proteins that are important in the pathogenesis of the virulent F. tularensis subsp. tularensis. We will use recently developed genetic tools to identify and disrupt genes encoding secreted proteins with the immediate goal of identifying proteins required for intracellular survival and replication in mouse macrophages. We anticipate that this work will identify several proteins of interest and will allow us to formulate hypotheses for the basis of further work. The work proposed in this application would contribute to the research goals of the NIAID biodefense research agenda for CDC category A agents such as Francisella tularensis. These immediate goals include; 1) the development of genetic systems to correlate differences in pathogenesis and virulence and 2) increased diversity of expertise in the study of tularemia. The research in this proposal will provide additional genetic methods for the study of F. tularensis biology and identify factors important for the pathogenesis of tularemia disease. This work may also contribute to better diagnostics, vaccine and drug development. Because of F. tularensis is a select agent and could be used a biological weapon, this research could potentially have a significant impact on public health in the future.