Humoral Immunity, Astrocyte Injury, and Demyelination in Neuromyelitis Optica

视神经脊髓炎的体液免疫、星形胶质细胞损伤和脱髓鞘

• 批准号: 10132323
• 负责人: Jeffrey L Bennett
• 金额: $ 37.71万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10132323
• 关键词: Address; Affect; Animal Diseases; Animal Model; Antibodies; Antibody Activation; Antigen Presentation; Astrocytes; Autoantibodies; Autoimmune Diseases; Autologous; Automobile Driving; B cell repertoire; B-Lymphocyte Subsets; B-Lymphocytes; BLR1 gene; Binding; Biological Assay; Biological Markers; Blindness; Blood - brain barrier anatomy; Brain; CNS Demyelinating Autoimmune Diseases; Caring; Catalogs; Cell Separation; Cells; Centers for Disease Control and Prevention (U.S.); Cerebrospinal Fluid; Complement; Complement-Dependent Cytotoxicity; Critical Pathways; Data; Demyelinations; Development; Diagnosis; Disease; Disease model; Disease remission; Endothelium; Epitopes; Eye; Eye diseases; Fostering; Funding; Histopathology; Humoral Immunities; ITGAX gene; IgG autoantibodies; Image; Immune; Immunoglobulin Class Switching; Immunoglobulin D; Immunoglobulin G; Immunoglobulin Variable Region; Immunoglobulins; In Vitro; Individual; Inflammatory; Injury; Intercellular adhesion molecule 1; Investigation; Knowledge; Lesion; Magnetic Resonance Imaging; Measures; Mediating; Memory B-Lymphocyte; Molecular; NF-kappa B; Neuraxis; Neurologic; Neuromyelitis Optica; Nuclear Translocation; Optic Nerve; Outcome; Paralysed; Pathogenicity; Pathology; Patients; Permeability; Physicians; Plasmablast; Population; Prevention; Production; Property; Recombinant Antibody; Recurrent disease; Relapse; Research; Retina; Risk; Role; Sampling; Serum; Severities; Severity of illness; Signal Pathway; Signal Transduction; Specificity; Spinal Cord; Stimulus; System; T-Lymphocyte; Technology; Testing; Therapeutic; Vascular Permeabilities; Visual; antibody-dependent cell cytotoxicity; aquaporin 4; base; brain endothelial cell; central nervous system injury; chemokine; clinically relevant; contrast enhanced; cytokine; cytotoxicity; design; disability; effective therapy; glucose-regulated proteins; in vivo; insight; novel; novel therapeutics; peripheral blood; prevent; relapse prediction; response; screening; tissue injury; translational study; water channel

Project Summary Neuromyelitis optica spectrum disorder (NMOSD) is a severe autoimmune disorder targeted against the aquaporin-4 (AQP4) water channel. Central nervous system (CNS) injury in NMOSD is initiated by the binding of AQP4 autoantibodies (AQP4-IgG) to target astrocytes and the activation of antibody effector functions; however, serum AQP4-IgG titers do not correlate with disease relapse or severity. Therefore, identifying factors that influence CNS access, intrathecal production, or effector function of AQP4-IgG is essential for understanding NMOSD pathognesis. Using single-cell sorting, recombinant antibody technology, and heavy-chain variable region repertoire analysis, we have reconstructed the intrathecal AQP4-IgG response in NMOSD, isolated anti-endothelial antibodies that activate brain microvascular endothelial cells (BMECs), and identified an expanded CD27-IgD- double negative (DN) memory B cell population clonally related to intrathecal AQP4-specific plasmablasts. We are now uniquely prepared to test our hypothesis that AQP4-IgG, autoantibodies against BMECs, and pro-inflammatory DN B cells act in concert to propel NMOSD activity and pathology. In Aim 1, we will examine the contribution of glucose regulated protein-78 (GRP78) autoantibodies to NMOSD disease activity. We will investigate the abundance and epitope specificity of NMOSD anti-GRP78 autoantibodies, evaluate the cell signaling pathways activated in BMECs, and measure their ability to increase vascular permeability in the brain and retina. In Aim 2, we will address how the binding specificity of individual anti-AQP4 autoantibodies affect lesion formation and CNS injury in NMOSD. We have identified multiple species of patient-derived AQP4-specific recombinant antibodies that produce distinct effects on target astrocytes. We will gauge AQP4 rAbs on their ability to activate complement dependent cytotoxicity, initiate antibody-dependent cell-mediated cytotoxicity, and modulate astrocyte chemokine and cytokine production in vitro. We will then evaluate how these parameters affect NMOSD lesion formation in animal models and disease activity in affected patients. And in Aim 3, we will analyze an expanded population of class-switched CD27-IgD- DN B cells in NMOSD patients and investigate their role in disease activity. The abundance and immunoglobulin repertoire of DN B cells will be compared during NMOSD relapse and remission, and we will assess the cytokine response and immunoglobulin production of NMOSD DN B cells in response to various stimuli. In addition, we will investigate the potential for NMOSD DN B cells to present AQP4 to autologous T cells. The results of these investigations will elucidate how AQP4-IgG accesses the CNS, how intrathecal B cells contribute to NMO lesion initiation, and how discrete subpopulations of AQP4-IgG contribute to CNS injury. The outcomes of our research will remove barriers to progress in the field and advance approaches to the diagnosis and treatment of NMOSD and other autoimmune disorders impacting the CNS and eye.

项目摘要 神经霉素炎选择性谱系障碍（NMOSD）是针对针对的严重自身免疫性疾病 Aquaporin-4（AQP4）水通道。 NMOSD中的中枢神经系统（CNS）损伤是由结合开始的 AQP4自身抗体（AQP4-IGG）的靶向星形胶质细胞和抗体效应子功能的激活； 但是，血清AQP4-IGG滴度与疾病复发或严重程度无关。因此，识别 影响CNS访问，鞘内产生或AQP4-IGG效应子功能的因素对于 了解NMOSD病情。使用单细胞分类，重组抗体技术和 重链变量区域曲目分析，我们重建了鞘内AQP4-IGG 在NMOSD中的反应，激活脑微血管内皮的孤立抗内皮抗体 细胞（BMEC），并鉴定出扩展的CD27-IGD-双阴性（DN）记忆B细胞种群 克隆与鞘内AQP4特异性浆膜相关。我们现在已经准备好测试我们的 假设AQP4-IGG，针对BMEC的自身抗体和促炎DN B细胞作用 促进NMOSD活动和病理学的音乐会。 在AIM 1中，我们将研究葡萄糖调节的蛋白质-78（GRP78）自身抗体对 NMOSD疾病活动。我们将研究NMOSD抗GRP78的丰度和表位特异性 自身抗体，评估在BMEC中激活的细胞信号通路，并测量其增加的能力 大脑和视网膜中的血管渗透性。在AIM 2中，我们将解决个体的约束性特异性 抗AQP4自身抗体会影响NMOSD病变形成和中枢神经系统损伤。我们已经确定了多个 患者衍生的AQP4特异性重组抗体的种类，对靶标产生明显的影响 星形胶质细胞。我们将根据其激活补体依赖的细胞毒性的能力来衡量AQP4 RABS，启动 抗体依赖性细胞介导的细胞毒性，并调节星形胶质细胞趋化因子和细胞因子的产生 体外。然后，我们将评估这些参数如何影响动物模型中的NMOSD病变形成 受影响患者的疾病活动。在AIM 3中，我们将分析扩大的类转换的人群 NMOSD患者的CD27-IGD-DN B细胞，并研究其在疾病活性中的作用。丰度和 在NMOSD复发和缓解期间，将比较DN B细胞的免疫球蛋白库，我们将 评估NMOSD DN B细胞的细胞因子反应和免疫球蛋白产生各种 刺激。此外，我们将研究NMOSD DN B细胞呈现AQP4的潜力 细胞。这些研究的结果将阐明AQP4-IGG如何访问CNS，鞘内b如何 细胞有助于NMO病变的启动，以及AQP4-IGG的离散亚群如何促进CNS 受伤。我们研究的结果将消除该领域进步的障碍，并进步 NMOSD和其他影响中枢神经系统和眼睛的自身免疫性疾病的诊断和治疗。