A novel blood-CSF adaptive immune response in Alzheimer's disease

项目摘要/摘要 阿尔茨海默病(AD)是一种无法治愈的神经退行性疾病，在这种疾病中，神经炎症日益严重 认识到发挥着关键作用。虽然先天炎症与阿尔茨海默病有关，但人们对此知之甚少 获得性免疫反应的贡献。本应用程序中提供的初步数据显示 以高分化CD8+T细胞增多为特征的AD外周免疫特征 记忆CD45RA+(TEMRA)细胞。值得注意的是，患者脑脊液中也存在CD8+TEMRA细胞 (CSF)和T细胞受体(TCR)测序显示其克隆性扩增，提示其抗原特异性。 这些适应性免疫细胞。TCR克隆和多肽筛选证实了克隆性细胞亚群的特异性 将AD-CSF TCR扩增为EB病毒BZLF1抗原。这些结果提供了第一个证据 受年龄相关影响的克隆性、抗原经验型T细胞在脑鞘内巡逻 神经退行性变。这个K99/R00应用程序将测试有害适应性免疫的新理论 对AD病理生物学有贡献的反应。在特定目的1，AD血-CSF T细胞克隆型将与 脑脊液生物标志物。这一方法将确定AD中特定的T细胞群以及这些细胞是否相关 疾病的严重性。在特定目标2中，将使用抗原鉴定筛选来检测自我/非我 S抗原促进AD患者T细胞克隆性增殖。这些检测可能会发现一个新的治疗靶点或 AD的生物标志物。特异性目标3将确定T细胞介导的神经元死亡和恢复的机制 在AD患者中，使用诱导神经元(IN)细胞与患者脑脊液CD8+T细胞共同培养。这些实验将评估 AD-CSF CD8+T细胞是否对EBV感染细胞和/或 BZLF1的分子模拟。候选人大卫·盖特博士在T细胞生物学方面拥有丰富的经验，并 花了十多年的时间研究AD。在该奖项的指导阶段，盖特博士的目标是推进他的 了解下一代测序分析、复杂的统计方法、抗原筛选、 培养方法和CRISPR基因编辑。Gate博士的导师和共同导师Tony Wyss-Coray博士和Dr。 马克·戴维斯分别在这些领域拥有全面的专业知识。他们将提供一个丰富的 为Gate博士发展成为神经免疫学研究领域杰出的独立研究员提供了良好的环境。AS 作为一名独立研究员，盖特博士将利用这一奖学金下的培训来全面和 定量评价T细胞分子成分与神经退行性变之间的相互作用。

Project Summary/Abstract Alzheimer’s disease (AD) is an incurable neurodegenerative disorder in which neuroinflammation is increasingly recognized to play a critical function. While innate inflammation has been implicated in AD, little is known about the contribution of the adaptive immune response. Preliminary data featured in this application demonstrate a peripheral immune signature of AD characterized by increased numbers of highly-differentiated CD8+ T effector memory CD45RA+ (TEMRA) cells. Strikingly, CD8+ TEMRA cells were also present in patient cerebrospinal fluid (CSF) and T cell receptor (TCR) sequencing indicated their clonal expansion, suggesting antigen specificity of these adaptive immune cells. TCR cloning and peptide screens demonstrated specificity of a subset of clonally expanded AD CSF TCRs to the Epstein-Barr virus (EBV) BZLF1 antigen. These results provide the first evidence of clonal, antigen-experienced T cells patrolling the intrathecal space of brains affected by age-related neurodegeneration. This K99/R00 application will test the novel theory of a detrimental adaptive immune response contributing to AD pathobiology. In Specific Aim 1, AD blood-CSF T cell clonotypes will be related to CSF biomarkers. This approach will determine specific T cell populations in AD and whether these cells relate to disease severity. In Specific Aim 2, antigen identification screens will be used to detect the self/non-self antigen(s) driving T cell clonal expansion in AD. These assays could uncover a novel therapeutic target or biomarker for AD. Specific Aim 3 will determine mechanisms of T cell-mediated neuronal death and resiliency in AD using induced neuronal (iN) cells co-cultured with patient CSF CD8+ T cells. These experiments will assess whether AD CSF CD8+ T cells mount cytotoxic effector responses to iN cells infected with EBV and/or to a molecular mimic of BZLF1. The candidate, Dr. David Gate, has extensive experience in T cell biology and has spent more than a decade studying AD. During the mentoring phase of this award, Dr. Gate aims to advance his knowledge in next-generation sequencing analysis, sophisticated statistical methods, antigen screening, iN culturing methods and CRISPR gene editing. Dr. Gate's mentor and co-mentor, Dr. Tony Wyss-Coray and Dr. Mark Davis, respectively, have comprehensive expertise in these areas. They will provide an enriching environment for Dr. Gate to develop as a prominent independent investigator in neuroimmunology research. As an independent investigator, Dr. Gate will leverage the training under this fellowship to comprehensively and quantitatively evaluate the interactions between T cell molecular components and neurodegeneration.