Toll-like Receptor 2, Vitamin D, and Extrapulmonary Tuberculosis

描述(申请人提供)：结核病(TB)是一种全球卫生紧急情况。世界卫生组织估计，2007年全球约有930万人被诊断出患有结核病，200万人死亡。结核病代表了一个有趣的悖论：据估计，世界上三分之一的人口感染了结核分枝杆菌，但只有5%-10%的感染者继续发展为结核病的临床表现。肺外结核可能是潜在的免疫妥协的标志。这位候选人的导师进行的研究表明，与之前有肺结核病和潜在结核分枝杆菌感染的人相比，有肺外结核病病史的人似乎有先天免疫缺陷。对既往肺外结核病患者的免疫反应的进一步研究可以提供线索，了解促使一个人从潜伏的结核分枝杆菌感染发展为临床疾病的免疫因素，并通过确定结核病疫苗的潜在靶点来促进结核病预防工作。新的证据表明，病原体识别受体称为Toll样受体(TLRs)，特别是TLR2和维生素D在结核分枝杆菌的免疫反应中发挥着重要作用。本研究的目的有二：(1)评估既往肺外结核病患者TLR2的数量和功能；(2)检测既往肺外结核病患者体内维生素D受体和维生素D介导的免疫激活的数量和功能。数量和功能的测量将使用聚合酶链式反应(PCR)和流式细胞术进行。结果将与既往肺结核患者、潜在结核分枝杆菌感染者和未感染结核分枝杆菌(PPD阴性)的健康对照组的结果进行比较。这些目标将使用巨噬细胞刺激和感染结核分枝杆菌的体外模型进行研究。 这位候选人在临床研究方面有很强的背景，获得了公共卫生硕士学位，并完成了几项结核病流行病学研究。这一奖项将支持她实现她的短期职业目标，即扩大她对结核分枝杆菌免疫反应的了解，同时在翻译免疫学研究方面建立坚实的基础，从而实现她成为一名独立和富有成效的翻译研究人员的长期目标。她的两个主要职业发展目标是：(1)加强她在免疫学和翻译研究方面的现有知识和获得新的专业知识；(2)在手稿和赠款中完善她的科学写作技能，以成功地通过RO1资助从NIH获得长期研究支持。这位候选人计划通过在实验室进行动手培训，参加几门高级免疫学和研究技能的教学课程，以及参加有利于与有共同兴趣的研究人员建立联系和合作的国内和国际科学会议，来增强她的免疫学和研究技能。她将通过科学写作的教学课程以及参加范德比尔特大学医学中心现有的写作研讨会和内部拨款审查机制来实现她的第二个目标。 她在范德比尔特大学医学中心的环境有利于她的抱负。她得到了她的主要导师蒂莫西·斯特林博士的支持，蒂莫西·斯特林博士是国际公认的结核病研究领域的专家，成功地指导了年轻的内科科学家，他们后来在学术医学领域从事了富有成效的研究生涯，并拥有支持她的研究努力的可用资源。她的另一位导师斯皮罗斯·卡拉姆斯博士是一位颇有成就的免疫学家，曾在免疫学实验室提供指导。她还得到了一个委员会的指导，该委员会由范德比尔特大学非常成功的基础研究人员和翻译研究人员组成，他们具有不同的研究兴趣，将帮助指导她的职业发展。候选人可获得的资源包括最先进的实验室设施和设备以及拥有先进技术的核心实验室。除了技术资源，候选人还可以在范德比尔特大学医学中心获得由临床和翻译科学奖(CTSA)提供的独特资源，如临床研究中心、范德比尔特临床和翻译研究所以及临床和翻译科学家发展计划(CTSA的教育组成部分)。该大学通过范德比尔特医生科学家发展计划(VPSD)支持这位候选人，这是一种竞争性的校内资助机制，是通往校外资助(即K奖)的“桥梁”。 目前的建议对结核病预防和疫苗学领域具有重要影响，这一奖项将使候选人能够在她坚实的临床结核病研究基础上发展新的翻译研究技能，并过渡到她职业生涯的下一个阶段。 项目简介：结核病是一种全球紧急情况。我们对人类对结核分枝杆菌的保护性免疫反应的了解是不完整的。免疫反应的进一步特征可能有助于识别从结核分枝杆菌感染进展到活动性疾病的人。这一知识将对结核病疫苗的开发和结核病预防工作产生重大影响。

DESCRIPTION (provided by applicant): Tuberculosis (TB) is a global health emergency. The World Health Organization estimated that approximately 9.3 million people worldwide were diagnosed with TB in 2007 and 2 million persons died. TB represents an interesting paradox: while one-third of the world's population is estimated to be infected with Mycobacterium tuberculosis, only 5-10% of infected persons go on to develop clinical manifestations of TB. Extrapulmonary TB is likely a marker of underlying immune compromise. The candidate's mentor has performed studies showing that persons with previous extrapulmonary TB appear to have an innate immune defect compared to persons with previous pulmonary TB and latent M. tuberculosis infection. Further study of the immune response of persons with previous extrapulmonary TB could provide clues to the immune factors that predispose a person to progress from latent M. tuberculosis infection to clinical disease and contribute to TB prevention efforts by identifying potential targets for TB vaccines. Emerging evidence suggests that pathogen recognition receptors called Toll-like receptors (TLRs), specifically TLR2, and vitamin D play a major role in the immune response to M. tuberculosis. In this study, two Specific Aims will be addressed: (1) to evaluate the quantity and function of TLR2 in persons with previous extrapulmonary TB and (2) to measure the quantity and function of the vitamin D receptor and vitamin-D mediated immune activation in persons with previous extrapulmonary TB. Measurements of quantity and function will be performed using polymerase chain reaction (PCR) and flow cytometry. Results will be compared to results obtained from persons with previous pulmonary TB, persons with latent M. tuberculosis infection, and healthy controls who are not infected with M. tuberculosis (PPD negative). The Aims will be investigated using an in vitro model of macrophage stimulation and infection with M. tuberculosis. The candidate has a strong background in clinical research, having obtained her Masters of Public Health and completed several TB epidemiologic studies. This award will support her while she achieves her short term career goal of expanding her knowledge of the immune response to M. tuberculosis, while establishing a strong foundation in translational immunologic research from which to attain her long term goal of becoming an independent and productive translational investigator. Her two primary Career Development Aims are: (1) to strengthen her existing knowledge and acquire new expertise in immunology and translational research and (2) to refine her scientific writing skills in manuscripts and grants to successfully compete for long-term research support through RO1 funding from NIH. The candidate plans to augment her immunology and research skills through hands-on training in the laboratory, participation in several didactic courses in advanced immunology and research skills, and attendance of national and international scientific meetings which will be favorable for networking and collaborating with investigators with shared interests. She will achieve her second Aim through didactic coursework in scientific writing as well as participation in available writing workshops and internal grant review mechanisms available at Vanderbilt University Medical Center. Her environment at Vanderbilt University Medical Center is conducive to her aspirations. She is supported by her primary mentor, Dr. Timothy Sterling, who is an internationally recognized expert in the area of TB research, has successfully mentored young physician-scientists who have gone on to productive research careers in academic medicine, and has the resources available to support her research efforts. Her co-mentor, Dr. Spyros Kalams, is an accomplished immunologist and has provided guidance in the immunology lab. She also is mentored by a committee consisting of highly successful basic and translational researchers at Vanderbilt with diverse research interests who will help guide her career development. The resources available to the candidate include state-of-the-art laboratory facilities and equipment as well as core laboratories with advanced technology. In addition to technical resources, the candidate has access to unique resources at Vanderbilt University Medical Center made possible by the Clinical and Translational Science Award (CTSA) such as the Clinical Research Center, the Vanderbilt Institute for Clinical and Translational Research, and the Clinical and Translational Scientist Development program, the educational component of the CTSA. The university has shown invested interest in the candidate by supporting her through the Vanderbilt Physician Scientist Development Program (VPSD), a competitive intramural funding mechanism that serves as a "bridge" to extramural funding (ie K award). The current proposal has important implications to the field of TB prevention and vaccinology, and this award will allow the candidate to build on her strong foundation of clinical TB research and develop new skills in translational research and transition to the next phase of her career. Project Narrative: Tuberculosis is a global emergency. Our understanding of the human protective immune response to Mycobacterium tuberculosis, the causative agent of tuberculosis is incomplete. Further characterization of the immune response could help identify persons who progress from M. tuberculosis infection to active disease. This knowledge would have significant implications for tuberculosis vaccine development and tuberculosis preventive efforts.