Department of Microbiology, Immunology and Biochemistry, The University of Tennessee Health Science Center, Memphis, Tennessee.
Curr Protoc Microbiol. 2020 Dec;59(1):e127. doi: 10.1002/cpmc.127.
Leptospirosis is a zoonotic disease caused by pathogenic Leptospira species that are maintained in sylvatic and domestic environments by transmission among rodents and other carriers. Humans become infected after contact of breached skin or mucosa with contaminated water or soil. Understanding persistent or sublethal infection in a host is critical for controlling human risk of exposure to pathogenic Leptospira. Animal models that recapitulate disease progression after infection via natural transmission routes are more appropriate for validation of vaccines and therapeutics. Furthermore, the ability to measure shedding of live Leptospira in urine of reservoir and carrier hosts can be used to develop new diagnostic assays and sensors to evaluate human risk of exposure. We developed inbred mouse models of Leptospirosis, that bypass survival as a criterion, in which we can analyze both pathogen and host factors affecting sublethal infection (<1 month), including shedding of Leptospira in urine. Mice are infected with pathogenic Leptospira using a physiologic route, and the clinical, histological, and molecular scores of disease are measured. Furthermore, the host immune response to Leptospira is evaluated. This mouse model also provides a tool in which to test fundamental hypotheses related to host-pathogen interactions and the immune mechanisms engaged in protective and pathogenic immune responses. © 2020 Wiley Periodicals LLC Basic Protocol 1: Culture and maintenance of virulent Leptospira Basic Protocol 2: Infection of mice through a physiologic route and collection of clinical scores and biological samples Basic Protocol 3: Analysis of pathogenesis after Leptospira infection.
钩端螺旋体病是一种由致病性钩端螺旋体引起的人畜共患病,这些螺旋体在森林和家庭环境中通过啮齿动物和其他载体之间的传播而得以维持。人类在接触受污染的水或土壤后,通过破损的皮肤或黏膜感染这种疾病。了解宿主中的持续性或亚致死性感染对于控制人类接触致病性钩端螺旋体的风险至关重要。能够重现自然传播途径感染后疾病进展的动物模型更适合疫苗和疗法的验证。此外,能够测量储菌宿主和带菌宿主尿液中活钩端螺旋体的脱落,可用于开发新的诊断检测和传感器,以评估人类接触的风险。我们开发了钩端螺旋体病的近交系小鼠模型,该模型不将生存作为标准,我们可以分析影响亚致死性感染(<1 个月)的病原体和宿主因素,包括尿液中钩端螺旋体的脱落。使用生理途径感染小鼠致病性钩端螺旋体,并测量疾病的临床、组织学和分子评分。此外,还评估了宿主对钩端螺旋体的免疫反应。这种小鼠模型还提供了一种工具,可以用来测试与宿主-病原体相互作用以及在保护性和致病性免疫反应中涉及的免疫机制相关的基本假设。 © 2020 威利父子公司 基本方案 1:培养和维持毒力钩端螺旋体 基本方案 2:通过生理途径感染小鼠并收集临床评分和生物样本 基本方案 3:分析钩端螺旋体感染后的发病机制。