Novel Antigen Identification for an enterotoxigenic E coli vaccine

项目概要/摘要 本项目的长期目标是开发一种安全、广泛保护的肠出血性大肠杆菌疫苗。 大肠杆菌（ETEC），是全球腹泻的常见原因。这个高度合作的项目将依赖于联合 资源，以及具有分子微生物学互补专业知识的研究人员的创新， 微生物发病机理、疫苗学、免疫学和先进的结构生物学， 研究ETEC疫苗开发的新靶点。 具体而言，该项目将： (1)检查ETEC的两种关键表面蛋白是否可以引起免疫回忆（记忆）反应， 被认为与持续保护免受感染有关 (2)从血液样本中获得的循环免疫细胞中开发单克隆抗体， 感染 (3)绘制出这些抗体与我们计划用于疫苗的两种表面蛋白质结合的位置 (4)确定我们计划用于疫苗的蛋白质的整体结构 (5)测试这些单克隆抗体中和蛋白质功能的能力以及细菌 传递毒素 (6)用这些蛋白质给小鼠接种疫苗，以测试对感染的保护作用 (7)测试小鼠抗体的活性，并绘制小鼠抗体与疫苗蛋白结合的位置 相对于人类抗体。 该项目将回答有关这两种蛋白质性质的基本问题，这些问题不仅与 ETEC而是分泌类似蛋白质的其他重要病原体。最重要的是，该项目预计 提供关键信息，可用于有效设计ETEC的多功能疫苗 提供广泛的长期保护。

Project Summary/Abstract The long-term objective of this project is to develop a safe and broadly protective vaccine for enterotoxigenic E. coli (ETEC), a common cause of diarrhea worldwide. This highly collaborative project will rely on the combined resources, and innovation of investigators with complementary expertise in molecular microbiology and microbial pathogenesis, vaccinology, immunology, and advanced state-of-the-art structural biology to investigate novel targets for ETEC vaccine development. Specifically, the project will: (1) examine whether two key surface proteins of ETEC can provoke immune recall (memory) responses that are thought to be involved in sustained protection against infection (2) develop monoclonal antibodies from circulating immune cells obtained from blood samples following natural infections (3) map where these antibodies bind on the two surface proteins that we plan to use in vaccines (4) determine the overall structure of the proteins that we plan to use in vaccines (5) test the ability of these monoclonal antibodies to neutralize protein function and the ability of the bacteria to deliver toxins (6) vaccinate mice with these proteins to test protection against infection (7) test the activity of antibodies from mice and map where antibodies from mice bind the vaccine proteins relative to the human antibodies. The project will answer fundamental questions about the nature of the two proteins that are relevant not only to ETEC but to other important pathogens that secrete similar proteins. Most importantly the project is expected to provide critical information that can be translated into efficient design of a multi-featured vaccine for ETEC that provides broad-based long-term protection.