Comprehensive Analysis of Borrelia burgdorferi Adhesins

伯氏疏螺旋体粘附素的综合分析

基本信息

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
项目类别：
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

项目摘要

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）：伯氏疏螺旋体是莱姆病的病原体，会造成持续感染，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多个部位传播和持续存在的能力表明，在感染过程中，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，许多记录或推定的粘附素已被鉴定，这表明不同粘附途径的协同作用使伯氏疏螺旋体能够在其特征性多相和多系统感染过程中协商连续步骤。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个有吸引力的模型。这种洞察力的缺乏部分是由于迄今为止确定的一些粘附素在结合活性方面表现出重叠，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，我们提出在没有潜在冗余机制的情况下对每种粘附素的特性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，每一位都拥有独特的专业领域，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，包括在非粘附、高传代菌株背景中生成功能获得性伯氏疏螺旋体突变体，以及测试菌株的体外粘附活性（目标 1，Leong 实验室）），与小鼠组织中的亲本菌株相比，对所有功能获得突变体的细菌负荷进行定量（目标 2，Coburn 实验室），并对每种突变体之间相互作用的性质进行活体显微镜检查活体小鼠的菌株和脉管系统（目标 3，Chaconas 实验室）。因此，该项目首次系统地研究了伯氏疏螺旋体的多种粘附素在生物体在免疫活性动物中建立播散性感染的能力中的作用。我们的长期目标是了解不同粘附素的生化活性如何共同发挥作用，使伯氏疏螺旋体克服宿主建立持续性、播散性感染的障碍。随着莱姆病在北半球的发病率持续扩大，无论是病例数量还是地理分布，这项工作可能会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。这个创新的多重 PI 项目汇集了具有独特专业知识的团队，将能够对大量具有明确生化活性的伯氏疏螺旋体蛋白质进行全面研究。这些蛋白质可能是开发候选疫苗或靶向治疗干预的绝佳候选蛋白。公共卫生相关性：莱姆病是北半球最流行的媒介传播疾病，对莱姆病常见地区的卫生系统造成重大负担。我们建议深入研究与哺乳动物组织成分结合的伯氏疏螺旋体蛋白如何对于细菌引起小鼠感染至关重要。该申请的三位主要研究人员共同组成了一支拥有独特专业知识的团队，将能够对大量伯氏疏螺旋体蛋白进行全面研究，这些蛋白具有独特的生化活性，并且可能是开发疫苗候选物或治疗干预目标的优秀候选者。