Recombinant murine IL-10 produced in situ by H. pylori

由幽门螺杆菌原位产生的重组鼠 IL-10

• 批准号: 6833463
• 负责人: KATHRYN A. EATON
• 金额: $ 6.96万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-15 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6833463
• 关键词: Helicobacter; gastritis; interleukin 10; laboratory mouse; molecular cloning; protein quantitation /detection; protein structure function; proteomics; recombinant proteins

DESCRIPTION (provided by applicant): H. pylori has been called the most common infectious disease of humans in the world today. Worldwide, between 50-100% of people are infected with H. pylori, but only a minority of those develop clinical signs of disease. Almost 2 decades of research have resulted in a general consensus that host immune response is a critical factor in determining the outcome of infection. Gastritis due to H. pylori is a T helper-1-mediated immune response associated with high levels of IFNgamma and low levels of IL-10 and other anti-inflammatory mediators. Understanding cytokine function in gastritis due to H. pylori has both diagnostic and therapeutic significance. First, identification of regulatory cytokines such as IL-10 or TGFbeta could lead to further understanding of regulatory pathways and development of therapies for those individuals with intractable infections. Second and perhaps equally important, such understanding could lead to the ability to identify individuals likely to be responders or non-responders. Individuals vary widely in their response to H. pylori, likely because of varying immunoreactivity of each individual host. Because of the strong association between IL-10 and immunoregulation of H. pylori responses, we have chosen to focus this pilot study on IL-10. Our overall goal is to test the hypothesis that recombinant murine IL-10 produced in situ by H. pylori ameliorates or prevents gastritis in mice. This goal will be accomplished in 2 specific aims: Aim 1: To engineer H. pylori to express recombinant murine IL- 10. Aim 2: To determine if in vivo expression leads to prevention or resolution of gastritis in response to recombinant bacteria or to wild-type H. pylori that co-colonize with recombinant strains. Briefly, we will construct recombinant H. pylori that expresses murine IL-10, demonstrate that the cytokine is expressed by bacteria in culture and in the mouse stomach, and determine if such expression ameliorates or prevents inflammation in a mouse model of severe gastritis. A successful outcome will not only determine the potential therapeutic role of IL-10 in gastritis due to H. pylori, but will also establish a model in which the roles of other cytokines and mediators can be determined, and host and bacterial interactions can be directly evaluated in vivo.

描述（由申请人提供）：幽门螺杆菌被称为当今世界上最常见的人类传染病。在全球范围内，有50-100％的人感染了幽门螺杆菌，但只有少数人出现疾病的临床迹象。将近20年的研究导致一般共识，即宿主免疫反应是确定感染结果的关键因素。幽门螺杆菌引起的胃炎是与高水平的IFNGAMMA和低水平IL-10和其他抗炎介质有关的T辅助辅助介导的免疫反应。 了解幽门螺杆菌引起的胃炎中细胞因子功能具有诊断和治疗意义。首先，鉴定IL-10或TGFBETA等调节性细胞因子可能会进一步了解那些患有棘手感染的人的调节途径和疗法的发展。其次，也许同样重要的是，这种理解可能会导致识别可能是反应者或无反应者的个人的能力。个体在对幽门螺杆菌的反应方面差异很大，这可能是由于每个宿主的免疫反应性变化。由于IL-10与幽门螺杆菌反应的免疫调节之间存在密切的关联，因此我们选择将这项初步研究集中在IL-10上。 我们的总体目标是检验以下假设：幽门螺杆菌原位产生的重组鼠IL-10可以改善或防止小鼠的胃炎。这个目标将以两个具体目标来实现： 目的1：要设计幽门螺杆菌以表达重组鼠IL-10。 目的2：确定体内表达是响应重组细菌还是与重组菌株共同殖民的野生型幽门螺杆菌的预防或分辨率。 简而言之，我们将构建表达鼠IL-10的重组幽门螺杆菌，表明细胞因子是通过培养物和小鼠胃中细菌表达的，并确定这种表达是否会改善或防止严重胃炎小鼠模型中的炎症。成功的结果不仅将确定IL-10在幽门螺杆菌引起的胃炎中的潜在治疗作用，而且还将建立一个模型，在该模型中，可以确定其他细胞因子和介质的作用，并且可以在体内直接评估宿主和细菌相互作用。