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The roles of Lyme spirochete adhesins in hematogenous dissemination

莱姆病螺旋体粘附素在血行传播中的作用

基本信息

批准号：
10570186
负责人：
George Chaconas
金额：
$ 71.46万
依托单位：
MEDICAL COLLEGE OF WISCONSIN
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-02-10 至 2027-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10570186
关键词：
Adhesives Animals Arthritis Bacteria Bacterial Adhesins Binding Biochemical Biology Blood Vessels Borrelia afzelii Borrelia burgdorferi Borrelia garinii CCL2 gene Cell Line Cells Chemosensitization Complex DBL Oncoprotein Endothelial Cells Endothelium Event Extracellular Matrix Gene Family Generations Genes Genetic Transcription Geography Health system Hematogenous Hour Infection Infectious Skin Diseases Integrin Binding Interleukin-10 Intravenous Invaded Investigation Joints Laboratories Left Ligands Lyme Disease Measures Mediating Modeling Monitor Mus Nature Order Spirochaetales Organism OspC protein Phase Prevention Principal Investigator Process Protein Array Research Personnel Role Skin TNF gene Testing Time Tissues Vector-transmitted infectious disease Visualization chronic infection cytokine decorin binding protein B genetic analysis genome wide screen in vivo inhibitor insight intravital microscopy meetings microbial colonization migration mouse model mutant neutrophil novel pathogenic microbe tick bite transmission process vector tick virtual

项目摘要

Upon transmission by a vector tick bite, Lyme disease spirochetes, primarily B. burgdorferi (Bb) in the US, establish a local skin infection, then disseminate to multiple tissues. Chronic infection by Bb is often associated with arthritis. Our laboratories have identified and/or characterized many Bb cell- or extracellular matrix (ECM)- binding adhesins using multiple approaches, overcoming the challenges of defining their roles in Bb biology. Our approaches include analyses of biochemical activities and generation of targeted mutants selectively defective for a single adhesive activity and analysis of the mutants in multiple murine infection models. To gain detailed mechanistic insight into interactions that may occur during Bb dissemination in vivo, following intravenous inoculation we used intravital microscopy to characterize vascular attachment and transmigration in skin and joint-proximal tissue. These studies revealed that adhesins BBK32 and VlsE account for virtually all of the transient Bb-endothelium binding occurring minutes after inoculation, termed “Meeting” interactions. A distinct set of adhesins, DbpB/A, OspC, and P66, mediate contacts required for invasion into extravascular joint space after 24 hours (hr), termed “Transmigrating” interactions. Using isogenic strain sets that have acquired or lost specific adhesive activities in multiple short-term and long-term infection models we showed roles for five of the six Meeting or Transmigrating adhesins in short-term tissue localization and/or long-term colonization in other murine infection models. While Transmigrating adhesins do not promote “Meeting” interactions, our discovery of enhanced adhesive capacity of the endothelium as infection progresses has helped clarify why different adhesins have roles at different stages of infection. Within hr, “Endothelial Activation” permits BBK32- and VlsE-independent (“Greeting”) interactions. After ~24 hr “Endothelial Potentiation” occurs, reflected by the ability of the joint vasculature to support Bb transmigration. Although both OspC and P66 function as Transmigrating adhesins, only P66, an integrin-binding adhesin that alters transcription in cultured endothelial cells, is also Potentiating, i.e., required to promote the rapid transmigration of a second Bb strain. Activation is mimicked by exogenous treatment of mice with several cytokines produced by infected mice, but potentiation is seen only with TNF-α, MCP-1 or IL-10. These findings reveal previously unrecognized steps that are critical for Bb spread and provide a means to distinguish roles for each adhesin in distinct infection stages: Meeting, Greeting, Potentiating, Transmigrating and Colonizing. In Aim 1 we will identify known adhesins that facilitate Greeting interactions; to better prioritize our adhesin list, we propose a genome-wide screen that may also identify novel adhesins. In Aim 2 we will clarify the roles of known (and, if applicable, novel) adhesins in endothelial Potentiation and Transmigration. Our use of rigorous genetic analyses in infection models from visualization of key interactions in vivo to quantitative analysis of Bb at different stages of infection will result in detailed understanding of a critical facet of Bb biology: dissemination.

经蜱叮咬传播后，莱姆病螺旋体，在美国主要是伯氏疏螺旋体（Bb），