LSU VETERINARY COBRE: PATHOGENESIS OF RICKETTSIA SP

路易斯安那州立大学兽医 COBRE：立克次体 SP 的发病机制

• 批准号: 8167884
• 负责人: Kevin R. Macaluso
• 金额: $ 19.98万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167884
• 关键词: Adult; Antigens; Blood capillaries; Cellular Structures; Computer Retrieval of Information on Scientific Projects Database; Coupled; Cues; Dermacentor; Event; Funding; Gene Expression; Grant; Immunoblotting; Individual; Infection; Institution; Larva; Liquid Chromatography; Metabolic; Nymph; Ovary; Pathogenesis; Proteins; Research; Research Personnel; Resources; Rickettsia; Rickettsia rickettsii; Salivary Glands; Source; Staging; Ticks; Time; Tissues; Two-Dimensional Gel Electrophoresis; United States National Institutes of Health; Vertical Disease Transmission; capillary; egg; feeding; novel; polyclonal antibody; protein expression; protein profiling; transmission process; vector

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Specific Aims. Aim 1. Characterize the rate at which SFG rickettsiae spread through individual ticks and throughout individual tick tissues. Uninfected Dermacentor variabilis will be exposed to Rickettsia rickettsii, R. montanensis, or R. bellii via capillary feeding. Parameters of SFG infection and transmission, including dispersion and multiplication, in individual tick tissues will be determined. A. Assess SFG rickettsial infection and subsequent tissue dissemination in the tick vector using quantitative real-time PCR. B. Quantify rickettsiae associated with horizontal (salivary gland) and vertical (ovary) transmission events in infected egg, larva, nymph, and adult stages. Aim 2. Identify and characterize uniquely expressed protein from three tick-borne rickettsiae in tick hosts during horizontal and vertical transmission events. Although not well understood, a metabolic union between other SFG rickettsiae and their tick hosts has been described. Rickettsia undergo a reactivation event changing in cell structure and vertebrate infectivity in association with host environmental cues coupled to bloodmeal acquisition. This aim will provide novel information concerning tick-borne rickettsiae in relation to global protein expression and antigenic profiles. A. Initial studies will compare protein profiles of SFG rickettsiae during transmission events within the tick tissues involved in horizontal (salivary gland) and vertical (ovary) transmission by two dimensional gel electrophoresis. B. Identify rickettsial antigens by immunoblot using polyclonal antibodies to tick-borne rickettsiae and liquid chromatography quadruple-time-of-flight (2DLC-QTOF) analysis. C. Compare observed protein expression to gene expression, as assessed by real-time PCR.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 具体目标。 目标 1. 描述 SFG 立克次体通过个体蜱虫和个体蜱组织传播的速率。未感染的变异皮肤蜱将通过毛细管喂养接触立克次体、山地皮肤蜱或贝氏立克次体。将确定 SFG 感染和传播的参数，包括个体蜱组织中的分散和繁殖。 A. 使用定量实时 PCR 评估蜱载体中的 SFG 立克次体感染和随后的组织传播。 B. 量化与受感染卵、幼虫、若虫和成虫阶段的水平（唾液腺）和垂直（卵巢）传播事件相关的立克次体。 目标 2. 鉴定并表征水平和垂直传播事件期间蜱宿主中三种蜱传立克次体的独特表达蛋白。尽管还不太清楚，但其他 SFG 立克次体与其蜱宿主之间的代谢结合已被描述。立克次体经历了细胞结构和脊椎动物感染性改变的再激活事件，这与宿主环境因素和血粉采集相关。这一目标将提供有关蜱传立克次体与整体蛋白质表达和抗原谱相关的新信息。 答：初步研究将通过二维凝胶电泳比较涉及水平（唾液腺）和垂直（卵巢）传播的蜱组织内传播事件期间 SFG 立克次体的蛋白质谱。 B. 使用蜱传立克次体多克隆抗体和液相色谱四重飞行时间 (2DLC-QTOF) 分析通过免疫印迹鉴定立克次体抗原。 C. 通过实时 PCR 评估，将观察到的蛋白质表达与基因表达进行比较。