High-throughput proteomic study of protective immunity in human cholera infection

描述(由申请人提供)：这是一份K08临床科学家导师研究职业发展奖培训计划的建议书。候选人里切尔·C·查尔斯博士最近完成了马萨诸塞州总医院和波士顿布里格姆妇女医院之间的联合培训项目中的传染病临床奖学金。她目前正在马萨诸塞州总医院感染科接受长期研究培训，并计划在世界资源有限的地区从事专注于输入性粘膜和肠道感染的独立生物医学调查员的职业生涯。本申请建议在马萨诸塞州总医院传染病科爱德华·T·瑞安博士的主要指导下，开展为期五年的K08培训计划。斯蒂芬·B·卡尔德伍德博士将在整个拟议项目的整个过程中提供共同指导。该计划包括讲授和实践培训，重点是高通量免疫蛋白质组学方法，并建立在与孟加拉国国际腹泻病研究中心(ICDDR，B)、哈佛蛋白质组学研究所和亚利桑那州立大学持续合作的基础上。 主要的项目重点将是将免疫蛋白质组学应用于人类霍乱感染的保护性免疫研究，霍乱感染在50多个国家仍是地方病。据估计，每年有300万至500万人罹患霍乱，导致约10万人死亡。尽管霍乱疫苗确实能诱导保护性免疫反应，但免疫是短暂的，大约持续6-24个月。相比之下，自然感染霍乱会导致持续数年或数十年的保护性免疫；然而，对霍乱保护性免疫反应的介体知之甚少。在这项应用中，学员将使用一个高通量的基于蛋白质的平台--核酸可编程蛋白质阵列，以(1)表征血清抗V-V。霍乱患者和接种者的霍乱免疫反应与基线血清体液抗V。霍乱患者家庭接触者的霍乱免疫与疾病保护；(2)粘膜抗V。用粘膜免疫标志物淋巴细胞抗体(ALS)检测霍乱弧菌的免疫应答；(3)鉴定抗霍乱弧菌。人野生型霍乱弧菌感染过程中霍乱菌记忆B细胞反应及相关抗-V。霍乱患者家庭接触者中的霍乱记忆B细胞免疫与疾病保护。通过高通量筛查确定的抗原随后将被更详细地检查，包括在免疫学验证性研究中。这些研究的结果可能为人类霍乱感染的保护性免疫提供重要的见解，并可能导致改进霍乱疫苗接种策略。K08奖项将为候选人提供科学研究方面的关键技能和经验，并促进她发展成为一名独立的内科科学家。

DESCRIPTION (provided by applicant): This is a proposal for a K08 Mentored Clinical Scientist Research Career Development Award training program. The candidate, Dr. Richelle C. Charles, recently completed an Infectious Disease clinical fellowship in the combined training program between Massachusetts General Hospital and the Brigham and Women's Hospital, Boston. She is now in a period of extended research training in the Division of Infectious Diseases at the Massachusetts General Hospital, and plans to pursue a career as an independent biomedical investigator focused on mucosal and enteric infections of import in resource-limited areas of the world. This application proposes a five-year K08 training program under the primary mentorship of Dr. Edward T. Ryan in the Division of Infectious Disease of the Massachusetts General Hospital. Dr. Stephen B. Calderwood, within the same division, will provide co-mentorship throughout the course of the proposed program. The program involves both didactic and practical training focused on high-throughput immuno-proteomic approaches, and builds upon ongoing collaborations with the International Center for Diarrheal Disease Research, Bangladesh (ICDDR,B), Harvard Institute of Proteomics, and Arizona State University. The major project focus will be the application of immuno-proteomics to the study of protective immunity in human cholera infection, an infection that remains endemic in over 50 countries. An estimated 3-5 million individuals develop cholera each year, resulting in approximately 100,000 deaths. Although cholera vaccines do induce a protective immune response, immunity is short-lived, lasting approximately 6-24 months. In comparison, natural infection with cholera results in protective immunity that lasts years or decades; however, the mediators of the protective immune response to cholera are poorly understood. In this application, the trainee will use a high-throughput protein-based platform, the Nucleic Acid Programmable Protein Array, to (1) characterize serum anti-V. cholerae immune responses in cholera patients and vaccinees, and correlate baseline serum humoral anti-V. cholerae immunity with protection from disease in household contacts of cholera patients; (2) characterize mucosal anti-V. cholerae immune response, using the antibody-in lymphocyte (ALS ) assay, a marker of mucosal immunity, and (3) characterize anti-V. cholerae memory B cell responses during human wild-type V. cholerae infection, and correlate anti-V. cholerae memory B cell immunity with protection from disease in household contacts of cholera patients. Antigens identified by high throughput screening will then be examined in more detail, including in immunologic confirmatory studies. The results of these studies could provide important insights into protective immunity of human cholera infection, and could lead to improved vaccination strategies against cholera. A K08 award would provide the candidate with critical skills and experience in scientific investigation, and facilitate her development into an independent physician scientist.