The Immune Response to F. tularensis in situ

对土拉弗氏菌的原位免疫反应

• 批准号: 6912410
• 负责人: JUDY M TEALE
• 金额: $ 27.08万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6912410
• 关键词: Francisella tularensis; NAD(P)H dehydrogenase; bactericidal immunity; bioterrorism /chemical warfare; cell migration; chemokine; chemokine receptor; gene expression profiling; host organism interaction; immune response; laboratory mouse; leukocyte activation /transformation; microarray technology; neutrophil; nitric oxide synthase; pathologic process; toll like receptor; virulence

DESCRIPTION (provided by applicant): The long-term goal of this project is to perform a careful kinetic analysis of the immune response to Francisella tularensis and the resulting pathology. The underlying hypothesis is that the innate immune response and/or early adaptive immune responses to pneumonic tularemia caused by F. tularensis subsp. tularensis (Schu4) is delayed or different compared to less virulent subspecies or attenuated mutants. The rationale is that comparing immune responses between virulent and less virulent strains will provide new insights into protective responses concentrating on the less virulent strains known to modulate the immune response. Three aims are proposed to compare Schu4 with targeted attenuated mutants. First, bacterial trafficking will be compared using labeled organisms followed by imaging in real time with the microPET and biodistribution of radioisotope and viable organisms. This will help determine whether differences in bacterial migration correlate with virulence as well as to identify the most important organs/tissues for study. Secondly, mice will be infected with Francisella strains/mutants (aerosol chamber) and sacrificed at various times post infection for removal of blood and relevant tissues defined in Aim 1. Initially, gene profiling will be done on isolated RNA from infected tissues by targeted macroarrays, thus focusing on immune related molecules important in the infection. Subsequently, in situ immunofluoresence approaches will be used, so that the pathogen, immune cells, and mediators can be identified at the same time. In addition, the extent of pathology will be assessed with time. Based upon our preliminary data, the third aim will concentrate on chemokines and the role of infiltrating neutrophils in disease progression. In collaboration with Project 2, we will also analyze TLR expression in situ. Information regarding virulence/protective mechanisms obtained in Projects 1-3 will interface with the 'humanized' transgenic model from Project 4. The data from this subproject, using real time imaging and in situ protocols, will provide a foundation for the disease processes associated with pneumonic tularemia. This project is therefore an integral part of the overall goal of this Program Project to understand the pathogenesis and host response to tularemia. (Cores C, B, and A required.)

描述（由申请人提供）：该项目的长期目标是对土拉弗朗西斯菌的免疫反应及其产生的病理学进行仔细的动力学分析。 基本假设是先天免疫反应和/或早期适应性免疫 对由土拉菌亚种引起的肺炎土拉热病的反应。 tularensis (Schu4) 与毒性较低的亚种或减毒突变体相比，延迟或不同。其基本原理是，比较强毒力和弱毒力菌株之间的免疫反应将为保护性反应提供新的见解，重点是已知可调节免疫反应的弱毒力菌株。提出了三个目标来将 Schu4 与靶向减毒突变体进行比较。首先，将使用标记的生物体对细菌运输进行比较，然后使用 microPET 进行实时成像以及放射性同位素和活生物体的生物分布。这将有助于确定细菌迁移的差异是否与毒力相关，以及确定用于研究的最重要的器官/组织。其次，小鼠将感染弗朗西斯氏菌菌株/突变体（气溶胶室），并在感染后不同时间处死，以去除目标 1 中定义的血液和相关组织。首先，将通过靶向宏阵列对从感染组织中分离的 RNA 进行基因分析，从而关注在感染中重要的免疫相关分子。随后，将使用原位免疫荧光方法，以便同时识别病原体、免疫细胞和介质。此外，病理的程度将随着时间的推移进行评估。根据我们的初步 根据数据，第三个目标将集中于趋化因子和浸润性中性粒细胞在疾病进展中的作用。与项目2合作，我们还将原位分析TLR表达。项目 1-3 中获得的有关毒力/保护机制的信息将与项目 4 中的“人源化”转基因模型相结合。该子项目的数据使用实时成像和原位协议，将为与肺炎兔热病。因此，该项目是该计划项目总体目标的一个组成部分，旨在了解兔热病的发病机制和宿主反应。 （需要核心 C、B 和 A。）