Decorin-binding proteins of Borrelia burgdorferi

描述（由申请人提供）：莱姆病仍然是一个主要的公共卫生问题，莱姆病的发病机制和免疫学的许多方面因此需要进一步研究。核心蛋白聚糖结合蛋白A（DbpA）是伯氏疏螺旋体（Borrelia burgdorferi）的一种膜脂蛋白，参与了B的寄生策略。通过在哺乳动物组织侵入期间作为细胞基质结合粘附素发挥作用而与伯氏螺旋体结合。DbpA也已成为最突出的新的人类莱姆病疫苗候选者。然而，DbpA的许多特征，如其时间表达，膜拓扑结构，在B中的作用。对伯氏菌毒力和作为保护性免疫原的总体效用仍然知之甚少。目前的研究解决了这些重要的信息差距。为此，本建议的具体目的是：（1）检查蜱传播的B的DbpA的时间表达模式。（2）研究蜱传B表达DbpA的膜拓扑结构。（3）构建强毒B.通过dbpA的插入失活，重点研究DbpA表达在B中的作用。进一步研究DbpA作为疫苗原在莱姆病小鼠模型中的总体功效。关于后者，重点将放在（a）纯化重组DbpA（作为免疫原）以保留其天然构象和（B）蜱侵染（B的攻毒途径）。burgdorferi）来模拟天然B。伯氏传播目前的提案代表了对DbpA在莱姆病发病机制中的作用及其作为人类莱姆病疫苗的潜力的全面研究。

DESCRIPTION (provided by applicant): Lyme disease continues to represent a major public health problem, and many aspects of Lyme disease pathogenesis and immunology thus warrant further investigation. Decorin-binding protein A (DbpA), a membrane lipoprotein of Borrelia burgdorferi, has been implicated in subserving the parasitic strategy of B. burgdorferi by functioning as a cell matrix-binding adhesin during mammalian tissue invasion. DbpA also has emerged as the most prominent new human Lyme disease vaccine candidate. However, many features of DbpA, such as its temporal expression, membrane topology, role in B. burgdorferi virulence, and overall utility as a protective immunogen, remain poorly understood. The current study addresses these important information gaps. To this end, the Specific Aims of this proposal are: (1) To examine the temporal expression pattern(s) of DbpA by tick-transmitted B. burgdorferi in the mammalian (mouse) host, (2) To examine the membrane topology of DbpA expressed by tick-transmitted B. burgdorferi in the mammalian (mouse) host, (3) To construct a DbpA-deficient mutant of virulent B. burgdorferi by insertional inactivation of dbpA, with emphasis on examining the role of DbpA expression in B. burgdorferi infectivity, virulence, and disease pathogenesis, and (4) To examine further the overall efficacy of DbpA as a vaccinogen in the mouse model of Lyme borreliosis. Regarding the latter, emphasis will be placed on (a) purifying recombinant DbpA (as immunogen) to preserve its native conformation and (b) tick infestation (challenge route for B. burgdorferi) of DbpA-immunized mice to mimic natural B. burgdorferi transmission. The current proposal represents a comprehensive study of the role of DbpA in both the pathogenesis of Lyme disease and its potential as a human Lyme disease vaccine.