Comprehensive Analysis of Borrelia burgdorferi Adhesins

伯氏疏螺旋体粘附素的综合分析

• 批准号: 8209007
• 负责人: George Chaconas
• 金额: $ 44.61万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8209007
• 关键词: Address; Adherence; Adhesions; Adhesives; Affect; Alleles; Animals; Area; Bacteria; Bacterial Adhesins; Bacteriophages; Binding; Biochemical; Bite; Blood Vessels; Borrelia burgdorferi; Cell-Matrix Junction; Cells; Characteristics; Cultured Cells; Development; Evaluation; Extracellular Matrix; Generations; Geographic Distribution; Goals; Health system; Heart; Human; Image; Immunocompetent; In Vitro; Infection; Infectious Skin Diseases; Integrins; Intravenous; Investigation; Joints; Knock-out; Knowledge; Label; Laboratories; Life; Lipoproteins; Lyme Disease; Mammalian Cell; Measures; Mediation; Microscopic; Mind; Modeling; Molecular; Mus; Nature; Nervous system structure; Order Spirochaetales; Organism; Pathogenesis; Pathway interactions; Phage Display; Phenotype; Play; Prevalence; Principal Investigator; Property; Protein Binding; Proteins; Reagent; Recombinant Proteins; Recombinants; Resolution; Role; Site; Skin; Specificity; Surface; Testing; Therapeutic Intervention; Ticks; Time; Tissues; Video Microscopy; Virulence; Work; base; cell type; functional group; gain of function; in vivo; innovation; insight; mutant; novel; public health relevance; tissue tropism; vaccine candidate; vector

DESCRIPTION (provided by applicant): Borrelia burgdorferi, a causative agent of Lyme disease, establishes persistent infection that can affect the joints, heart, skin, and nervous system. The abilities of this spirochete to disseminate and persist in a variety of sites in the mammalian host indicate that interactions with mammalian cells and extracellular matrix occur continually during infection. In fact, many documented or putative adhesins have been identified, suggesting that the concerted action of diverse adhesion pathways enables B. burgdorferi to negotiate successive steps during its characteristic multiphasic and multisystemic infection. Unfortunately, an important gap in our current knowledge is the lack of any mechanistic information to inform this attractive model. This paucity of insight is due in part to the fact that some of the adhesins identified to date display an overlap in binding activities, a factor that confounds straightforward analysis of any single adhesion pathway. Here we propose a comprehensive in vitro and in vivo analysis of the properties and function of each adhesin in the absence of potentially redundant mechanisms. To this end, this application brings together three principal investigators, each with a unique area of expertise, to address this critical knowledge gap. The three laboratories will employ state of the art approaches, including generation of gain-of-function B. burgdorferi mutants in a non-adherent, high-passage strain background and testing of the strains for in vitro adhesion activities (Aim 1, Leong lab), quantification of bacterial burdens for all gain-of-function mutants in comparison to the parental strain in mouse tissues (Aim 2, Coburn lab), and intravital microscopic examination of the nature of the interaction between each of the strains and the vasculature in living mice (Aim 3, Chaconas lab). This project therefore constitutes the first systematic examination of the roles of the diverse adhesins of B. burgdorferi in the ability of the organism to establish disseminated infection in immunocompetent animals. Our long-term goal is to understand how the biochemical activities of different adhesins function together to allow B. burgdorferi to overcome host barriers to the establishment of persistent, disseminated infection. As Lyme disease prevalence continues to expand in the Northern hemisphere, in terms of both case numbers and geographic distribution, this work may illuminate how B. burgdorferi causes Lyme disease in humans and persists in its animal reservoirs. This innovative multiple PI project brings together teams with unique expertise that will allow comprehensive investigations of a large set of B. burgdorferi proteins with defined biochemical activities. These proteins may be excellent candidates for development as vaccine candidates or for targeting as therapeutic interventions. PUBLIC HEALTH RELEVANCE: Lyme disease is the most prevalent vector-borne illness in the northern hemisphere, and a significant burden on the health system in regions in which it is common. We propose to delve into how Borrelia burgdorferi proteins that bind to mammalian tissue components are critical for the bacteria to cause infection in mice. The three principal investigators of this application together are a team with unique expertise that will allow comprehensive investigations of a large array of B. burgdorferi proteins that have unique biochemical activities and may be excellent candidates for development as vaccine candidates or targeting for therapeutic interventions.

描述（由申请人提供）：伯氏疏螺旋体，莱姆病的病原体，建立持续感染，可影响关节，心脏，皮肤和神经系统。这种螺旋体在哺乳动物宿主的各种部位传播和持续存在的能力表明，在感染期间，与哺乳动物细胞和细胞外基质的相互作用不断发生。事实上，许多文献记载的或推测的粘附素已经被确定，这表明多种粘附途径的协同作用使伯氏疏螺旋体在其特征性的多相和多系统感染过程中能够协调连续的步骤。不幸的是，在我们目前的知识中，一个重要的差距是缺乏任何机制信息来告知这个有吸引力的模型。这种缺乏洞察力的部分原因是迄今为止发现的一些粘附素在结合活动中表现出重叠，这是一个混淆任何单一粘附途径的直接分析的因素。在这里，我们提出了一个全面的体外和体内分析的性质和功能的每个粘连素在没有潜在的冗余机制。为此，该申请汇集了三位主要研究人员，每个人都有独特的专业领域，以解决这一关键的知识差距。这三个实验室将采用最先进的方法，包括在非黏附的高传代菌株背景下产生功能获得型伯氏螺旋体突变体，并测试菌株的体外黏附活性（目标1，Leong实验室），将所有功能获得型突变体的细菌负荷与小鼠组织中的亲本菌株进行比较（目标2，Coburn实验室），以及活体显微检查每一种菌株与活小鼠血管系统之间相互作用的性质（目标3，查科纳斯实验室）。因此，该项目首次系统地研究了伯氏疏螺旋体的各种粘附素在机体在免疫能力强的动物中建立播散性感染的能力中所起的作用。我们的长期目标是了解不同粘附素的生化活性如何共同起作用，使伯氏疏螺旋体克服宿主障碍，建立持久的、播散性感染。随着莱姆病在北半球的流行继续扩大，在病例数和地理分布方面，这项工作可能阐明伯氏疏螺旋体如何在人类中引起莱姆病并在其动物宿主中持续存在。这个创新的多PI项目汇集了具有独特专业知识的团队，将允许对具有明确生化活性的大量伯氏疏螺旋体蛋白质进行全面调查。这些蛋白质可能是开发候选疫苗或靶向治疗干预措施的极好候选者。