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

项目摘要/摘要 创伤性脑损伤(TBI)是一个主要的健康问题；每年有250万美国人遭受脑损伤，其代价是 每年800亿美元。很少有治疗方法可以减少脑外伤的长期认知后遗症，而且只有 康复的选择有限。脑损伤会导致初级结构损伤，继而是第二阶段， 这涉及到先天免疫系统和适应性免疫系统的激活。伴有小胶质细胞的神经炎 脑挫裂伤的发病机制已被认为是一种主要因素。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CD3抗体鼻腔给药 单抗诱导下调节小胶质细胞活化的抗炎免疫反应 在多发性硬化症动物模型中。其机制与鼻腔抗CD3抗体定位于颈淋巴有关。 诱导IL-10分泌(CD4+LAP+和CD4+FoxP3+)调节性T细胞迁移的结节 并抑制小胶质细胞的激活。鼻腔抗CD3治疗在颅脑损伤中是一个未被探索的领域，而 Tregs调节脑损伤小胶质细胞的机制在很大程度上还不清楚。Saef Izy博士，一位训练有素的 有急性脑损伤研究背景的神经强化学家假设鼻腔抗CD3代表 一种独特的、临床适用的治疗脑损伤的免疫调节方法。伊兹博士密切合作 与他的主要导师Howard Weiner博士一起，研究鼻腔抗CD3抗体对脑外伤预后的影响 老鼠。他的初步数据显示，鼻腔抗CD3增加了CD4+Tregs和IL 10的表达 减少受控皮质撞击伤(CCI)后7天脑内小胶质细胞的激活。它还提高了 损伤后1个月的行为结果。在这份K08提案中，伊兹博士将确定鼻腔注射的效果 抗CD3抗体对CCI后远期组织病理学和行为结局的影响(目标1)。他将调查 损伤后小胶质细胞、效应T细胞和调节性T细胞的反应，并研究Tregs对小胶质细胞的影响 CCI后的体外炎症反应(目标2)。他将研究鼻腔抗CD3对IL-10/IL-10的影响 使用携带IL-10Rflx/FloxTMEM119CreETR2的C57BL6/J小鼠在小胶质细胞中的10R信号转导 他莫昔芬作用后小胶质细胞表达IL-10受体。他还将扮演(S) Tregs在体内外通过中和转化生长因子-β和IL-4产生的其他抗炎细胞因子和 研究它们对CCI后行为学和小胶质细胞炎症反应的影响(目标3)。伊兹博士的职业目标是 为了更好地了解创伤性脑损伤后适应性免疫反应与先天免疫系统的接口。 这项提案的成功完成将使人们深入了解Tregs调节 颅脑损伤后的小胶质细胞反应及新型免疫治疗药物的发现 结果。建议的K08应用程序利用了Izy博士的导师在免疫学方面的专业知识，MICE 急性脑损伤模型和生物信息学为他提供了更多的知识和经验 有必要成为美国国立卫生研究院资助的脑外伤神经免疫学独立研究人员和专家。

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.