VIRULENCE ASSOCIATED PROTEINS OF BORRELIA BURGDORFERI

描述（改编自申请人的摘要）：疏螺旋体属 莱姆病的病原体，广义伯氏菌， 可能进展为持续性感染的多系统疾病 以显著的发病率为特征。这个项目的总体目标是 建议是鉴定和表征相关的强毒株 (VSA)外膜（OM）蛋白质，无论是外膜- 跨越（Oms）和脂蛋白，作为保护性免疫原。 研究人员的长期目标是将保护性免疫力 引起的致病机制。具体来说，这 研究提出：（1）纯化和表征28和35千道尔顿的 (kDa)B强毒株相关（VSA）OM蛋白。burgdorferi 感染性分离株的特异性抗体28和29的本土形式 将从源自B的OM囊泡中纯化35 kDa OM蛋白。 Burgdorferi及其氨基末端或内部肽序列 测定这些序列将被用来识别 通过筛选B来编码这些蛋白质。伯氏基因组学 数据库;（2）表征四个强毒株相关（VSA） B抗原。从噬菌体λ表达中鉴定出伯氏螺旋体 图书馆我们将确定差异表达的程度， 毒力与无毒力B中的VSA抗原。burgdorferi和评价 这些抗原是否在感染的动物中表达， 温度调节;和（3）测试这些OM VSA抗原， 他们赋予保护性免疫的能力。重组VSA蛋白 将被过度生产、纯化并用于免疫小鼠。天然 将VSA OM 28和35 kDa蛋白掺入脂质体中， 产生的蛋白脂质体用于免疫小鼠。免疫动物 然后用传染性B进行攻击。burgdorferi。血清特异性 将测试保护性免疫原，以确定这些 抗体提供被动保护，抗体是否介导 粘附抑制或优先杀死体外培养的毒力 B。burgdorferi以补体依赖的方式。测定 不同B的保护能力。表达的伯氏抗原 相对于在体外培养期间表达的那些，宿主将 促进疫苗鸡尾酒的发展， B的非克隆环境分离株。burgdorferi。而且这些 研究将提供深入了解由VSA介导的致病机制 B抗原。burgdorferi。

DESCRIPTION (Adapted from the Applicant's Abstract): Borrelia burgdorferi sensu lato, the etiologic agent of Lyme disease, causes a multi-system disorder that may progress into a persistent infection characterized by significant morbidity. The overall goal of this proposal is to identify and characterize virulent strain associated (VSA) outer membrane (OM) proteins, both those that are outer membrane- spanning (Oms) and lipoproteins, that function as protective immunogens. The investigators' long-term goal is to link protective immunity elicited by these proteins to pathogenic mechanisms. Specifically, this study proposes to: (1) Purify and characterize a 28 and 35 kilodalton (kDa) virulent strain associated (VSA) OM proteins of B. burgdorferi that are specific to infectious isolates. The native forms of the 28 and 35 kDa OM proteins will be purified from OM vesicles derived from B. burgdorferi and their amino terminal or internal peptide sequences determined. These sequences will then be used to identify the genes encoding these proteins by screening the B. burgdorferi genomic database; (2) Characterize four virulent strain associated (VSA) antigens of B. burgdorferi identified from a phage lambda expression library. We will determine the extent of differential expression of the VSA antigen in virulent versus avirulent B. burgdorferi and evaluate whether these antigens are either expressed in infected animals or temperature regulated; and (3) Test these OM VSA antigens identified for their ability to confer protective immunity. Recombinant VSA proteins will be overproduced, purified, and used to immunize mice. The native VSA OM 28 and 35 kDa proteins will be incorporated into liposomes and the resulting proteoliposomes used to immunize mice. Immunized animals will then be challenged with infectious B. burgdorferi. Serum specific for protective immunogens will be tested to determine whether these antibodies confer passive protection and whether antibodies mediate adherence inhibition or preferentially kill in vitro cultivated virulent B. burgdorferi in a complement dependent manner. Determination of the protective abilities of different B. burgdorferi antigens expressed in the host relative to those expressed during in vitro cultivation, will facilitate in the development of a vaccine cocktail to protect against non-clonal, environmental isolates of B. burgdorferi. Furthermore, these studies will provide insight into pathogenic mechanisms mediated by VSA antigens of B. burgdorferi.