Novel Antigen Identification for an enterotoxigenic E coli vaccine

项目摘要/摘要 该项目的长期目标是开发一种安全且具有广泛保护性的肠毒素E。 大肠埃希氏菌(ETEC)，全球腹泻的常见原因。这个高度协作的项目将依赖于 在分子微生物学和分子微生物学领域具有互补专业知识的研究人员的资源和创新 微生物发病机制，疫苗学，免疫学和先进的最先进的结构生物学 研究ETEC疫苗开发的新靶点。 具体而言，该项目将： (1)检测ETEC的两个关键表面蛋白是否能激发免疫回忆(记忆)反应， 被认为参与了对感染的持续保护 (2)从天然血样中获得的循环免疫细胞制备单抗 感染 (3)绘制这些抗体与我们计划在疫苗中使用的两种表面蛋白结合的位置 (4)确定我们计划用于疫苗的蛋白质的总体结构 (5)检测这些单抗中和蛋白质功能的能力，以及细菌中和蛋白质的能力 投放毒素 (6)用这些蛋白质免疫小鼠，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图 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，这两种蛋白质不仅与 ETEC，但对其他分泌类似蛋白质的重要病原体。最重要的是，该项目是预期的 为ETEC提供可转化为高效设计多功能疫苗的关键信息 这提供了广泛的长期保护。

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.