Comprehensive Analysis of Borrelia burgdorferi Adhesins

伯氏疏螺旋体粘附素的综合分析

• 批准号: 8063374
• 负责人: George Chaconas
• 金额: $ 47.14万
• 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/8063374
• 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; 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）：莱姆病的病因Borlelia Burgdorferi建立了持续的感染，可以影响关节，心脏，皮肤和神经系统。这种螺旋体的能力在哺乳动物宿主的各种部位传播和持续存在表明，与哺乳动物细胞和细胞外基质的相互作用在感染期间不断发生。实际上，已经确定了许多有记录的或推定的粘附素，这表明各种粘附途径的一致动作使B. Burgdorferi能够在其特征性的多相和多系统感染期间协商连续的步骤。不幸的是，我们当前知识的一个重要差距是缺乏任何机械信息来告知这种有吸引力的模型。这种洞察力的匮乏部分是由于以下事实：迄今为止确定的一些粘合剂在结合活动中表现出重叠，这是对任何单个粘附途径进行直接分析的因素。在这里，我们提出了在没有潜在冗余机制的情况下对每种粘附素的性质和功能进行全面的体外和体内分析。为此，该应用程序汇集了三位主要研究人员，每个研究人员都有独特的专业知识，以解决这一关键知识差距。 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活小鼠中每种菌株与脉管系统之间相互作用的性质（AIM 3，Chaconas Lab）。因此，该项目构成了B. burgdorferi在生物体在免疫能力动物中建立传播感染的能力中各种粘附素的作用的首次系统检查。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，以使B. burgdorferi克服宿主的障碍，以建立持续的，传播的感染。就病例数和地理分布而言，随着莱姆病的患病率在北半球继续扩展，这项工作可能会阐明B. burgdorferi如何引起人类的莱姆病，并在其动物储层中持续存在。这个创新的多个PI项目将具有独特专业知识的团队汇集在一起​​，可以全面调查具有定义的生化活动的大量B. burgdorferi蛋白。这些蛋白质可能是作为疫苗候选者开发的出色候选者或作为治疗干预措施的靶向候选者。 公共卫生相关性：莱姆病是北半球最普遍的媒介传播疾病，并且是常见地区的卫生系统负担很大。我们建议深入研究与哺乳动物组织成分结合的疏螺旋体burgdorferi蛋白如何对细菌引起小鼠感染至关重要。该应用程序的三位主要研究人员是一个具有独特专业知识的团队，将允许对拥有独特生物化学活动的大量B. burgdorferi蛋白质进行全面研究，并且可能是作为疫苗候选者开发或针对治疗干预措施的出色候选者。