Diagnosis and Pathobiology of Emerging Enterohepatic Helicobacter spp. in Mice

新兴肠肝螺杆菌的诊断和病理学。

• 批准号: 8308332
• 负责人: JAMES G FOX
• 金额: $ 42.2万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-01 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8308332
• 关键词: Address; Anti-Inflammatory Agents; Anti-inflammatory; Bacterial Antigens; Biochemical; Biological Assay; Chronic Hepatitis; Clinical; Colitis; Collaborations; Collection; Colon; Colon Carcinoma; Communities; Development; Diagnosis; Disease; Disease Progression; Enteral; Enterocolitis; Exclusion; Failure; Gastrointestinal Diseases; Genes; Genetic; Genome; Genomics; Helicobacter; Hepatic; Hepatitis; Hepatobiliary; Hepatocarcinogenesis; Human; Human Microbiome; Immune; Immune response; Immunocompromised Host; Immunodeficient Mouse; In Vitro; Inbred C57BL Mice; Inbred Mouse; Infection; Inflammation; Inflammatory Bowel Diseases; Institutes; Interleukin-10; Intestines; Laboratories; Laboratory Animals; Liver; MSC gene; Malignant neoplasm of liver; Mediating; Modeling; Molecular; Molecular Target; Morbidity - disease rate; Mus; Names; National Center for Research Resources; Nuclear; Oral; Organism; Primary carcinoma of the liver cells; Research; Ribosomal RNA; Rodent; Role; Route; Scientist; Screening procedure; Serological; Specific Pathogen Frees; Testing; Tumor Cell Line; Tumor Promoters; United States National Institutes of Health; Virulence; Virulence Factors; adaptive immunity; adenoma; aged; antimicrobial; base; candidate identification; cytokine; gastrointestinal; genome sequencing; germ free condition; in vitro Model; in vitro testing; in vivo; mortality; mouse model; novel; pathogen; prevent; prototype; reproductive; transmission process

DESCRIPTION (provided by applicant): We are excited about the opportunity to respond to the NCRR RFA-RR-10-005 "Pathobiology of Emerging Pathogens in Laboratory Animals". During the past 15 years, 8 novel enterohepatic Helicobacter spp. (EHS) in mice have been formally named, and 9 additional clusters (based on 16S rRNA sequences) of novel species have been cultured and identified in our lab. We have collected, what we believe, is the largest EHS culture collection available worldwide and are poised to characterize these murine strains, both to facilitate their diagnosis and also to establish their pathogenic potential. H. hepaticus, the prototype EHS, causes a persistent enteric infection and is prevalent in academic mouse colonies throughout the world. H. hepaticus infection causes liver cancer, is known to be a strong tumor promoter in several models of hepatocarcinogenesis, and induces inflammatory bowel disease in immunocompromised mice. The presence of EHS has confounded a number of studies, can contaminate tumor cell lines, causes significant morbidity and mortality in immunodeficient mice and selected strains of immune-competent mice, and has been recently associated with reproductive failures in mouse colonies. A 2007 study conducted by us indicated that 85% of mice used in academic settings worldwide are infected with one or more EHS. However the majority of EHS, we and others have identified, are not easily cultured, do not have specific molecular or serological assays for diagnosis, nor is it known whether and under what conditions these H. spp. have pathogenic potential. We therefore intend to develop sensitive and specific molecular assays to diagnose EHS in mice. We anticipate the development of a mini-microarray chip for use by the research community to detect EHS and prevent their colonization in established H. spp. free colonies. Because of the utility of the IL10-/- in our lab and others in using H. hepaticus to dissect etiopathogenesis of IBD, we will employ this model in screening novel H. spp. for pathogenic potential as well as determine the putative role of virulence genes in newly sequenced H. spp. These results will be compared and validated with in vitro tests we have used to predict virulence potential in H. hepaticus. The specific aims of the project are: 1) To provide morphologic, genetic, and biochemical characterization of novel enterohepatic Helicobacter species (EHS) and develop species specific assays to diagnose EHS isolated from mice; 2) To define the relationship of in vitro virulence factors of murine EHS to in vivo pathogenic potential in the IL10-/- mouse model and to determine how the hosts' immune responses influence disease progression; and 3) To identify and characterize virulence determinants in the genome sequences of 3 enteric Helicobacter spp. isolated from rodents and humans with clinical disease, as well as those of H. hepaticus and determine if these genes are operable in inducing IBD in C57BL IL10-/- mice. To accomplish these aims we have assembled a team of scientists who have successfully collaborated together for the last decade to study the etiopathogenesis of H. hepaticus gastrointestinal disease in mice.

描述（由申请人提供）：我们很高兴有机会回应实验动物中新兴病原体的NCRR RFA RRR-10-005“新兴病原体的病理学”。在过去的15年中，有8种新型肠肝杆菌属。 （EHS）在小鼠中已被正式命名，并且在我们的实验室中培养并鉴定出了新的物种的另外9个（基于16S rRNA序列）。我们认为，我们已经收集了全世界最大的EHS培养物品，并准备表征这些鼠标菌株，既可以促进其诊断，又要确定其致病潜力。 H. Hepaticus（原型EHS）引起了持续的肠道感染，并且在世界各地的学术老鼠殖民地中都普遍存在。 H.肝癌感染引起肝癌，在几种肝癌发生模型中是强烈的肿瘤启动子，并在免疫功能低下的小鼠中诱导炎症性肠病。 EHS的存在使许多研究混淆，可能污染肿瘤细胞系，在免疫缺陷的小鼠中引起明显的发病率和死亡率，并选择了免疫能力小鼠的菌株，并且最近与小鼠菌落的生殖失败有关。我们进行的一项2007年的研究表明，全世界在学术环境中使用的小鼠中有85％感染了一个或多个EHS。但是，我们和其他人已经确定的大多数EHS都不容易培养，没有特定的分子或血清学测定法以进行诊断，也不知道这些H. spp是否以及在什么条件下。具有致病潜力。因此，我们打算开发敏感和特定的分子测定，以诊断小鼠的EHS。我们预计开发了一个迷你微阵列芯片，用于研究界使用，以检测EHS并防止其在既定的H. spp中的殖民化。免费殖民地。由于我们的实验室中IL10 - / - 在使用H. h.paticus剖析IBD的疗法发生方面的实用性，因此我们将采用该模型来筛选新型H. spp。为了致病潜能，并确定毒力基因在新测序的H. spp中的推定作用。这些结果将通过体外测试进行比较和验证，我们用来预测肝螺旋杆菌中的毒力潜力。该项目的具体目的是：1）提供新型肠肝杆菌物种（EHS）（EHS）的形态，遗传和生化特征，并开发特定物种的测定，以诊断从小鼠分离的EHS； 2）定义鼠EHS体外毒力因子与IL10 - / - 小鼠模型中体内致病潜力的关系，并确定宿主的免疫反应如何影响疾病进展； 3）识别和表征3个肠形螺旋杆菌属的基因组序列中的毒力决定因素。从患有临床疾病的啮齿动物和人类以及肝螺旋杆菌的啮齿动物和人类中分离出来，并确定这些基因是否可以在诱导C57BL IL10 - / - 小鼠中诱导IBD中均可操。为了实现这些目标，我们组建了一个科学家团队，他们在过去的十年中成功合作，研究小鼠肝癌胃肠道疾病的疗法发生。