Analysis of pgf 54 Members in Lyme Disease Serodiagnosis

莱姆病血清学诊断中pgf 54成员分析

• 批准号: 7054063
• 负责人: James A. Carroll
• 金额: $ 20.88万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7054063
• 关键词: Borrelia; Ixodes; Lyme disease; SDS polyacrylamide gel electrophoresis; acidity /alkalinity; bacteria infection mechanism; bacterial proteins; bacterial vaccines; biomarker; body temperature; clinical research; communicable disease diagnosis; cooperative study; diagnosis design /evaluation; disease /disorder onset; early diagnosis; enzyme linked immunosorbent assay; gene expression; host organism interaction; human subject; laboratory mouse; latent bacterial disease; matrix assisted laser desorption ionization; patient oriented research; polymerase chain reaction; protein structure function; serology /serodiagnosis; vaccine development; vaccine evaluation; vector vaccine; western blottings

DESCRIPTION (provided by applicant): Borrelia burgdorferi, the etiologic agent of Lyme disease, is maintained in an infectious cycle that involves the transmission of the bacterium from an Ixodes tick to a mammalian host. During transmission the spirochetes encounter numerous alterations in environmental stimuli, and respond to these alterations in the environment by modifying the expression of genes that may prove important in adaptation, infection and persistence. Many of the genes that are differentially expressed reside on the many of the linear (lp) and circular (cp) plasmids that can be isolated from this spirochete. To date, the function of many of these pH and temperature-regulated membrane proteins has not been determined. Of interest are several members of paralogous gene family (pgf) 54 that localize to plasmid lp54 that are up-regulated in vitro when spirochetes are exposed to a temperature and pH (37 degrees C and pH 7.0) that mimic the environment of the attached, fed tick or mammal. Likewise, many of these pgf 54 members are down regulated by laboratory conditions that mimic the free-living, unfed tick (23 degrees C and pH 8.0). Moreover, we have determined that sera from mice infected via tick-borne transmission recognize nineteen borrelial membrane proteins, two of which are members of pgf 54. In Specific Aim 1 we intend to assess using the proteins encoded by the pgf 54 members as diagnostic markers of early, early-disseminated, and late stage Lyme disease. In Specific Aim 2 we will determine the efficacy of using recombinant proteins encoded by pgf 54 as vaccine candidates. Furthermore, in Specific Aim 3 we will identify additional pH-regulated membrane proteins expressed by B. burgdorferi during tickborne infection of mice that may be of diagnostic interest.

描述（由申请人提供）：莱姆病的病因学药Borrelia burgdorferi保持在传染性周期中，涉及细菌从ixodes向哺乳动物宿主的ixodes传播。在传播过程中，螺旋体遇到了环境刺激的许多改变，并通过修改可能证明对适应，感染和持久性重要的基因表达来应对环境中的这些改变。许多差异表达的基因都存在于许多可以从该螺旋体中分离出来的线性（LP）和圆（CP）质粒。迄今为止，尚未确定许多这些pH和温度调节的膜蛋白的功能。有趣的是，当螺旋体暴露于温度和pH（37摄氏度和pH 7.0）时，它们位于质粒LP54的几个成员（pGF）54，它们在体外被上调，模拟附着，喂养tick虫或哺乳动物的环境。同样，这些PGF 54名成员中的许多成员都受到模仿自由生活的未滴答的实验室条件（23摄氏度和pH 8.0）的调节。此外，我们已经确定，通过tick传播传播感染的小鼠的血清识别出了19个骨膜蛋白，其中两个是PGF 54的成员。在特定目标1中，我们打算使用PGF 54成员编码的蛋白质评估，该蛋白是PGF 54成员作为早期，早期诊断，早期释放的Lyme病的诊断标记。在特定目标2中，我们将确定使用由PGF 54作为候选疫苗的重组蛋白的功效。此外，在特定的目标3中，我们将确定B. burgdorferi在可能具有诊断兴趣的小鼠感染期间表达的其他pH调节的膜蛋白。