The Role of TNF in Breaking B Cell Tolerance

Quach博士的中心目标是获得新的技能，这将使她成为一名系统免疫学家。拟议 这项研究结合了鼠和人的研究，以确定TNF缺乏破坏耐受性的机制。 所获得的知识将指导未来为自身免疫性患者设计个性化药物的努力。 候选人：Quach博士是Feinstein医学研究所（FIMR）的讲师。通过博士学位 在博士后工作中，她专注于人类B细胞的发育和调控。拟议 职业发展计划将建立在她以前的经验基础上，有四个培训目标，以提高她的轨迹 成为一名独立的研究者：1）获得利用小鼠模型的专业知识; 2）精通 在计算生物学; 3）获得洞察力进行临床导向的研究项目，和4）建立一个 数据和方法的基础，以生成预测模型。 导师/环境：柯博士和她的导师Anne Davidson博士组建了一个强大的团队， 顾问和合作者指导她完成拟议的培训和研究活动。拟议 该项目利用FIMR的知识、研究和临床设施以及可用资源 通过她的外部顾问和合作者，史蒂芬·克莱因斯坦博士和伊纳基·桑兹博士。FIMR致力于 通过内部赠款和网络和教育机会支持初级教师。博士 当当地无法提供最佳培训时，Quach将参加国家研讨会/讲习班。 研究：在接受TNF抑制剂（TNFi）治疗的患者中诱导自身抗体和自身免疫是良好的- 然而，TNFi诱导B细胞耐受性破坏的机制尚未确定。在 在人类中，TNFi通过破坏生发中心（GC）的形成来影响B和T细胞的稳态， 用于高亲和力抗原特异性抗体生产和自身反应性B细胞的阴性选择。同样在 在小鼠中，TNF信号传导缺陷阻止GC形成，诱导产生TFH和CD 4 +IL-17的细胞扩增， 并改变自身抗体谱。这项研究提出，TNF缺乏，加上第二次诱导， 刺激，通过减少负性GC B细胞信号传导和增强T 效应细胞活性。为了检验这一假设，第一个目的是利用2种不同背景的TNF缺陷型小鼠， 用TLR 9激动剂和NZM2328.TNFR1/2双缺陷小鼠诱导的自身反应性Sle1.TNF-/-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 来加强这个过程。第二个目标将解决TNFi治疗患者中的类似问题， 荧光试剂检测和分离ANA反应性B细胞结合下一代测序 技术.表型和功能研究的结合用于确定T细胞的影响。的 本研究的结果将阐明TNFi对调节B细胞耐受性的作用，并改善我们的免疫功能。 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.