LSU VETERINARY COBRE: PATHOGENESIS OF RICKETTSIA SP

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 具体目标。 目标1.表征SFG立克次体通过个体蜱和个体蜱组织传播的速率。未感染的变异革蜱将暴露于立克次氏体、立克次氏体。montanensis或R.通过毛细血管进食。将确定单个蜱组织中SFG感染和传播的参数，包括分散和增殖。 A.使用定量实时PCR评估SFG立克次体感染和随后在蜱虫载体中的组织传播。 B。定量立克次体在感染卵、幼虫、若虫和成虫阶段的水平（唾液腺）和垂直（卵巢）传播事件。 目标2.鉴定和表征三种蜱传立克次体在蜱宿主中水平和垂直传播事件中独特表达的蛋白质。虽然还没有很好地理解，其他SFG立克次体和它们的蜱宿主之间的代谢联盟已经被描述。立克次体经历一个再激活事件，改变细胞结构和脊椎动物感染性与宿主环境线索耦合血粉收购。这一目标将提供有关蜱传立克次体的全球蛋白表达和抗原谱的新信息。 A.初步研究将通过二维凝胶电泳比较SFG立克次体在涉及水平（唾液腺）和垂直（卵巢）传播的蜱组织内传播事件期间的蛋白质谱。 B。通过使用蜱传立克次体多克隆抗体的免疫印迹和液相色谱四重飞行时间（2DLC-QTOF）分析鉴定立克次体抗原。 C.将观察到的蛋白质表达与通过实时PCR评估的基因表达进行比较。

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.