Targeting CNS Neuroinflammation in Traumatic Brain Injury by Nasal Anti-CD3

项目总结/摘要 创伤性脑损伤（TBI）是一个主要的健康问题;每年有250万美国人遭受TBI， 每年800亿美元。很少有疗法能减少TBI的长期认知后遗症， 康复的选择有限。脑损伤导致原发性结构损伤，随后是继发性阶段， 其涉及先天免疫系统和适应性免疫系统的激活。神经炎症伴小胶质细胞 参与已经被确定为TBI发病机制的主要贡献者。然而，仍然没有 有效的免疫治疗来调节损伤后的小胶质细胞反应。抗CD 3经鼻给药 单克隆抗体诱导抗炎免疫应答，下调小胶质细胞活化 在多发性硬化症的动物模型中。其机制涉及鼻抗CD 3定位于颈淋巴 在淋巴结中诱导分泌IL-10的（CD 4 + T细胞+和CD 4 + FoxP 3+）调节性T细胞（T细胞）迁移 并抑制小胶质细胞的激活鼻腔抗CD 3治疗是TBI的一个未探索领域， 在TBI中，TBI调节小胶质细胞的机制在很大程度上是未知的。Saef Izzy医生，一位训练有素的 一位有急性脑损伤研究背景的神经重症监护医师假设，鼻抗CD 3代表 一种治疗TBI的独特的、临床适用的免疫调节方法。伊兹医生与 与他的主要导师，霍华德韦纳博士，研究鼻抗CD 3对TBI结果的影响， 小鼠他的初步数据显示，鼻用抗CD 3抗体可增加CD 4 + T细胞亚群和IL 10的表达， 减少受控皮质撞击损伤（CCI）后7天脑中的小胶质细胞活化。它也提高 伤后1个月的行为结果。在这个K 08提案中，Izzy博士将确定鼻腔给药的影响。 抗CD 3对CCI后长期组织病理学和行为结果的影响（目的1）。他将调查 损伤后小胶质细胞、效应细胞和调节性T细胞的反应，并研究TdR对小胶质细胞、效应细胞和调节性T细胞的影响。 CCI后的体外炎症反应（目的2）。他将研究鼻用抗CD 3抗体对IL-10/IL-10的影响。 使用携带IL-10 Rflox/ILTMEM 119 CreETR 2的C57 BL 6/J小鼠在小胶质细胞中的10 R信号传导， 在他莫昔芬给药后小胶质细胞上表达IL-10受体。他还将阐述以下方面的作用： 由TclA通过在体外和体内中和TGF-β和IL-4产生的其他抗炎细胞因子， 研究它们对CCI后行为和小胶质细胞炎症反应的影响（目的3）。伊兹医生的职业目标是 以更好地了解TBI后适应性免疫反应如何与先天免疫系统相互作用。 成功完成这一提案将提供深入了解的机制，其中Tendon调节 TBI后的小胶质细胞反应和识别新的基于免疫的治疗方法，以改善患者的 结果。拟议中的K 08应用程序利用了Izzy博士的导师在免疫学、小鼠免疫学和免疫学方面的专业知识。 急性脑损伤模型和生物信息学为他提供了额外的知识和经验 有必要成为一个独立的NIH资助的调查员和专家在TBI的神经免疫学。

PROJECT SUMMARY / ABSTRACT Traumatic brain injury (TBI) is a major health problem; 2.5 million Americans sustain TBI each year at a cost of 80 billion dollars annually. Few therapies reduce long-term cognitive sequelae of TBI, and there are only limited options for rehabilitation. Brain injury causes a primary structural injury followed by a secondary phase, which involves the activation of the innate and adaptive immune systems. Neuroinflammation with microglia’s involvement has been identified as a major contributor to the pathogenesis of TBI. However, there is still no effective immune therapy to modulate the microglial response post-injury. Nasal administration of anti-CD3 monoclonal antibody induces an anti-inflammatory immune response that down-regulates microglial activation in animal models of multiple sclerosis. The mechanism involves localization of nasal anti-CD3 to cervical lymph nodes where it induces IL-10-secreting (CD4+LAP+ and CD4+FoxP3+) regulatory T cells (Tregs) that migrate to the brain and inhibit microglial activation. Nasal anti-CD3 therapy is an unexplored area in TBI, and the mechanisms by which Tregs modulate microglia in TBI are largely unknown. Dr. Saef Izzy, a trained neurointensivist with a background in acute brain injury research, hypothesized that nasal anti-CD3 represents a unique, clinically applicable immunomodulatory approach for the treatment of TBI. Dr. Izzy worked closely with his primary mentor, Dr. Howard Weiner, to investigate the effects of nasal anti-CD3 on TBI outcomes in mice. His preliminary data showed that nasal anti-CD3 increases CD4+Tregs and IL 10 expression and reduces microglial activation in the brain 7 days after controlled cortical impact injury (CCI). It also improves behavioral outcomes at 1-month post-injury. In this K08 proposal, Dr. Izzy will determine the effects of nasal anti-CD3 on long-term histopathological and behavioral outcomes after CCI (Aim 1). He will survey the microglial, effector, and regulatory T cell responses after injury and study the effects of Tregs on microglial inflammatory response in vitro after CCI (Aim 2). He will investigate the effect of nasal anti-CD3 on IL-10/IL- 10R signaling in microglia using a C57BL6/J mouse harboring IL-10Rflox/floxTMEM119CreETR2, which does not express the IL-10 receptor on microglia after tamoxifen administration. He will also delineate the role(s) of other anti-inflammatory cytokines produced by Tregs by neutralizing TGF-β and IL-4 in vitro and in vivo and study their effects on behavior and microglial inflammatory response post-CCI (Aim 3). Dr. Izzy's career goal is to better understand how the adaptive immune response interfaces with the innate immune system after TBI. Successful completion of this proposal will provide insight into the mechanisms by which Tregs modulate the microglial response after TBI and the identification of novel immune-based therapeutics to improve patients’ outcomes. The proposed K08 application leverages Dr. Izzy’s mentor’s expertise in immunology, mouse models of acute brain injury, and bioinformatics to provide him with the additional knowledge and experience necessary to become an independent NIH-funded investigator and expert in the neuroimmunology of TBI.