The Role of TNF in Breaking B Cell Tolerance

夸赫博士的中心目标是获得新技能，从而确立她作为系统免疫学家的地位。建议数 这项研究结合了老鼠和人类的研究，以确定肿瘤坏死因子缺乏打破耐受性的机制。 所获得的知识将指导未来为自身免疫患者设计个性化药物的努力。 候选人：夸赫博士是范斯坦医学研究所(FIMR)的讲师。通过她的博士学位 和博士后工作，她专注于人类B细胞的发展和调节。建议数 职业发展计划将以她以前的经验为基础，制定四个培训目标，以提升她的职业生涯 成为一名独立的研究人员：1)获得利用鼠标模型的专业知识；2)熟练掌握 在计算生物学方面；3)在进行面向临床的研究项目方面获得洞察力，以及4)建立 数据和方法的基础，以生成预测性模型。 导师/环境：Quach博士和她的导师Anne Davidson博士组建了一个强大的团队 指导她完成拟议的培训和研究活动的顾问和合作者。建议数 项目利用了FIMR现有的智力、研究和临床设施以及可用的资源 通过她的外部顾问和合作者史蒂文·克莱因斯坦博士和稻崎·桑兹博士。FIMR致力于 通过内部助学金以及网络和教育机会支持初级教员。Dr。 当当地没有最佳培训时，Quach将参加国家研讨会/讲习班。 研究：使用肿瘤坏死因子抑制剂(TNFi)治疗的患者诱导自身抗体和自身免疫效果良好 已知；然而，TNFi诱导B细胞耐受破坏的机制尚不确定。在……里面 人类，TNFi通过破坏生发中心(GC)的形成而影响B和T细胞的动态平衡 用于高亲和力抗原特异性抗体的产生和自身反应性B细胞的阴性选择。同样，在 小鼠，肿瘤坏死因子信号缺陷阻止GC形成，诱导TFH和CD4+IL-17产生细胞扩张， 并改变自身抗体图谱。这项研究提出，肿瘤坏死因子缺乏与第二种诱导 刺激，通过减少负的GC B细胞信号和增强T细胞来损害GC B细胞的选择 效应器细胞活动。为了验证这一假设，第一个目的是利用两个不同背景的肿瘤坏死因子缺陷小鼠， 自身反应性Sle1.TLR9激动剂诱导的肿瘤坏死因子-/-小鼠和NZM2328.TNFR1/2双缺陷小鼠 确定GC B细胞中改变B细胞选择的信号缺陷的机制，以及T细胞如何帮助 以加强这一进程。第二个目标是用一种新的方法解决接受TNFi治疗的患者的类似问题 荧光试剂检测分离ANA反应性B细胞结合下一代测序 技术表型和功能研究相结合的方法被用来确定T细胞的影响。这个 这项研究的结果将阐明TNFi在调节B细胞耐受性方面的作用，并改善我们的 了解当GC形成异常时，免疫系统如何调节B细胞耐受性。

Dr. Quach's central goal is to acquire new skills that will establish her as a systems immunologist. The proposed research combines murine and human studies to identify mechanisms by which TNF deficiency breaks tolerance. The gained knowledge will guide future efforts in designing personalized medicine for autoimmune patients. Candidate: Dr. Quach is an instructor at the Feinstein Institute for Medical Research (FIMR). Through her Ph.D and post-doctoral work, she focused on the development and regulation of B cells in humans. The proposed career development plan will build upon her previous experience with four training goals to enhance her trajectory toward becoming an independent investigator: 1) gain expertise in utilizing mouse models; 2) become proficient in computational biology; 3) gain insights into conducting clinically oriented research projects, and 4) build a foundation of data and methodologies to generate predictive models. Mentors/Environment: Dr. Quach and her mentor, Dr. Anne Davidson, have assembled a strong team of advisors and collaborators to guide her through the proposed training and research activities. The proposed project utilizes the intellectual, research and clinical facilities available at the FIMR and the resources available through her external advisors and collaborators, Dr. Steven Kleinstein and Dr. Inaki Sanz. FIMR is committed to support junior faculty members through internal grants and opportunities for networking and education. Dr. Quach will attend national seminars/workshops when optimal training is not available locally. Research: The induction of autoantibody and autoimmunity in patients treated with TNF inhibitors (TNFi) is well- known; however, the mechanism by which TNFi induce breach of B cell tolerance is yet to be determined. In humans, TNFi affect B and T cell homeostasis via disruption of germinal center (GC) formation which is pivotal for high affinity antigen-specific antibody production and negative selection of autoreactive B cells. Similarly, in mice, TNF signaling deficiency prevents GC formation, induces TFH and CD4+IL-17 producing cell expansion, and alters autoantibody profiles. This study proposes that TNF deficiency, together with a second inducing stimulus, compromises GC B cell selection via reduction of negative GC B cell signaling and enhancement of T effector cell activities. To test this hypothesis, the first aim utilizes TNF deficient mice of 2 different backgrounds, autoreactive Sle1.TNF-/- mice induced with a TLR9 agonist and NZM2328.TNFR1/2 double deficient mice, to determine the mechanism for the signaling defect in GC B cells that alters B cell selection, and how T cells help to enhance this process. The second aim will address similar questions in TNFi treated patients using a novel fluorescent reagent to detect and isolate ANA reactive B cells combined with next generation sequencing technology. A combination of phenotyping and functional studies is used to determine T cells' influences. The results from this study will elucidate the effects of TNFi on regulating B cell tolerance and improve our understanding of how the immune system regulates B cell tolerance when GC formation is abnormal.