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)：伯氏疏螺旋体是莱姆病的病原体，在感染循环中保持不变，包括将细菌从硬蜱传播到哺乳动物宿主。在传播过程中，螺旋体会遇到环境刺激的许多变化，并通过改变可能被证明在适应、感染和持久性方面重要的基因的表达来响应环境中的这些变化。许多差异表达的基因存在于许多可以从螺旋体中分离出来的线性(Lp)和环状(Cp)质粒上。到目前为止，许多这些调节pH和温度的膜蛋白的功能还没有确定。令人感兴趣的是平行基因家族(PGF)54的几个成员，它们定位于质粒lp54，当螺旋体暴露在模拟附着的、喂食的扁虱或哺乳动物的环境的温度和pH(37摄氏度和pH 7.0)时，在体外上调表达。同样，这些PGF54成员中的许多成员受到实验室条件的下调，这些条件模仿自由生活的未喂养的扁虱(23摄氏度和PH8.0)。此外，我们已经确定，通过扁虱传播感染的小鼠的血清识别19种支气管膜蛋白，其中两种是PGF54的成员。在特定目标1中，我们打算使用由PGF54成员编码的蛋白质作为早期、早期播散性和晚期莱姆病的诊断标记物进行评估。在具体目标2中，我们将确定使用由PGF54编码的重组蛋白作为候选疫苗的效果。此外，在特定的目标3中，我们将鉴定可能具有诊断意义的伯氏芽孢杆菌在壁虱传播感染小鼠过程中表达的其他pH调节的膜蛋白。