Defining the role of B cells in modulating CNS inflammation in multiple sclerosis

摘要 衰弱的中枢神经系统(CNS)疾病多发性硬化症(MS)的特征是大量 外周免疫细胞与轴突脱髓鞘和损伤重合，导致疼痛和受损 协调。B细胞在多发性硬化发病机制中的作用已经显现，因为研究表明B细胞定向 治疗大大减少了新的炎性脱髓鞘病变的形成。此外，大多数人 多发性硬化症患者的病变既有B细胞也有反应性小胶质细胞。虽然这些发现 证实了B细胞的致病作用，目前尚不清楚哪些B细胞功能有助于脱髓鞘 发炎的中枢神经系统。此外，全身性B细胞清除会严重损害患者的免疫功能， 确定开发更有针对性的治疗方法的必要性。新的证据表明多发性硬化症的B细胞 患者是促炎细胞因子的有力调节者，这些异常的B细胞可以调节中枢神经系统 当它们进入中枢神经系统时，先天免疫系统。这项应用的研究将选择性地局部消融B 新型caspase-9介导的细胞凋亡转基因工具在中枢神经系统细胞中的实验研究 自身免疫性脑脊髓炎(EAE)MS动物模型初步数据显示靶向CNS B细胞 EAE症状出现后，小胶质细胞反应性降低，并伴有髓鞘减少 损伤和功能障碍。中枢神经系统B细胞来源的小胶质细胞炎症信号通路耗竭 将使用先进的磷酸流式细胞术技术对EAE动物进行评估。脱髓鞘和 大脑和脊髓中的重新髓鞘形成将在这个强大的、新的动物模型中进一步研究，以测试 中枢神经系统B细胞切除可防止进一步的髓鞘损伤并促进髓鞘修复。多发性硬化B细胞在骨肉瘤中的作用 神经炎症将通过刺激人类小胶质细胞分泌多发性硬化症患者来源的B细胞来确定 分析小胶质细胞转录和翻译炎症活性的影响因素。B细胞诱导 小胶质细胞的激活将在人类多发性硬化症脑损伤中得到证实。总之，拟议的研究将提供 对MS复杂免疫病理的新见解，涉及CNS B细胞和 小胶质细胞。这些研究的完成将带来新的免疫调节治疗途径， 在不影响患者免疫的情况下减轻多发性硬化症的疾病负担。 拟议的培训计划是由乔治华盛顿大学学院的罗伯特·H·米勒博士赞助的。 医学与健康科学。总体目标是为PI，Julie J Ahn，建立一个坚实的基础 为她在神经免疫学和神经免疫领域的独立科学家职业生涯做好准备 神经炎。研究金培训计划包括以下目标：1)增进科学知识 和技术技能，结合神经科学和免疫学，2)通过口头陈述研究成果 报告和手稿，3)指导和领导年轻科学家，4)负责任地进行研究。

Summary The debilitating central nervous system (CNS) disease multiple sclerosis (MS), is characterized by an influx of peripheral immune cells that coincides with axonal demyelination and damage, resulting in pain and impaired coordination. The role of B cells in MS pathogenesis has emerged as studies showed that B cell directed therapies substantially reduce the formation of new inflammatory demyelinating lesions. Further, the majority of lesions from patients with established MS harbor B cells as well as reactive microglia. While these findings corroborate a pathogenic role for B cells, it is unclear which B cell function contributes to demyelination within the inflamed CNS. In addition, systemic B cell elimination can severely immunocompromise patients, establishing the need to develop more targeted therapies. Emerging evidence suggests that B cells of MS patients are potent regulators of pro-inflammatory cytokines, and these abnormal B cells can regulate the CNS innate immune system when they enter the CNS. Studies in this application will selectively and locally ablate B cells in the CNS using a novel caspase 9-mediated apoptosis transgenic tool in the well-studied experimental autoimmune encephalomyelitis (EAE) animal model of MS. Preliminary data indicate that targeting CNS B cells after the onset of symptoms in EAE reduces microglial reactivity and is accompanied by decreased myelin damage and functional impairment. Inflammatory signaling pathways in microglia from CNS B cell depleted EAE animals will be evaluated using advanced phospho-flow cytometry techniques. Demyelination and remyelination in the brain and spinal cord will be further studied in this powerful, novel animal model to test if CNS B cell ablation prevents further myelin injury and promotes myelin repair. The role of MS B cells in neuroinflammation will be determined by stimulating human microglia with MS patient-derived B cell secreted factors and analyzing transcriptional and translational inflammatory activity of microglial cells. B cell induced microglial activation will be confirmed in human MS brain lesions. Together, the proposed studies will provide novel insights into the complex immunopathology of MS that involves the interaction of CNS B cells and microglia. Completion of these studies will lead to novel immunomodulatory therapeutic avenues that reduce the disease burden of multiple sclerosis without immunocompromising the patient. The proposed training plan is sponsored by Dr. Robert H. Miller at the George Washington University School of Medicine and Health Sciences. The overall goal is to build a strong foundation for the PI, Julie J Ahn, to prepare her for a successful career as an independent scientist in the fields of neuroimmunology and neuroinflammation. The fellowship training plan includes the following goals: 1) enhance scientific knowledge and technical skills that integrate neuroscience and immunology, 2) present research findings through oral presentations and manuscripts, 3) mentor and lead young scientists, and 4) conduct research responsibly.