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: 9898380
• 负责人: Jeffrey L Bennett
• 金额: $ 38.88万
• 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/9898380
• 关键词: Address; Affect; Animal Diseases; Animal Model; Antibodies; Antibody Activation; Antigen Presentation; Astrocytes; Autoantibodies; Autoimmune Diseases; Autoimmune Process; 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; 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.

项目概要 视神经脊髓炎谱系障碍 (NMOSD) 是一种严重的自身免疫性疾病，针对 水通道蛋白 4 (AQP4) 水通道。 NMOSD 中的中枢神经系统 (CNS) 损伤是由结合引发的 AQP4 自身抗体 (AQP4-IgG) 靶向星形胶质细胞并激活抗体效应功能； 然而，血清 AQP4-IgG 滴度与疾病复发或严重程度无关。因此，识别 影响 AQP4-IgG 的 CNS 通路、鞘内产生或效应器功能的因素对于 了解 NMOSD 发病机制。利用单细胞分选、重组抗体技术， 重链可变区库分析，我们重建了鞘内AQP4-IgG NMOSD 中的反应，分离的抗内皮抗体可激活脑微血管内皮细胞 细胞 (BMEC)，并鉴定出扩展的 CD27-IgD-双阴性 (DN) 记忆 B 细胞群 与鞘内 AQP4 特异性浆母细胞克隆相关。我们现在已经做好了独特的准备来测试我们的 假设 AQP4-IgG、针对 BMEC 的自身抗体和促炎 DN B 细胞在 音乐会推动 NMOSD 活动和病理学。 在目标 1 中，我们将研究葡萄糖调节蛋白 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 BMEC 中激活的细胞信号通路，并测量其增强能力 大脑和视网膜的血管通透性。在目标 2 中，我们将解决个体的结合特异性如何 抗 AQP4 自身抗体影响 NMOSD 病变形成和 CNS 损伤。我们已经确定了多个 患者来源的 AQP4 特异性重组抗体种类，对靶点产生独特的影响 星形胶质细胞。我们将评估 AQP4 rAb 激活补体依赖性细胞毒性的能力，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，并 受影响患者的疾病活动。在目标 3 中，我们将分析扩大的职业转换群体 NMOSD 患者中的 CD27-IgD-DN B 细胞并研究其在疾病活动中的作用。丰富度和 将在 NMOSD 复发和缓解期间比较 DN B 细胞的免疫球蛋白库，我们将 评估 NMOSD DN B 细胞响应各种因素的细胞因子反应和免疫球蛋白产生 刺激。此外，我们将研究 NMOSD DN B 细胞向自体 T 呈递 AQP4 的潜力 细胞。这些研究的结果将阐明 AQP4-IgG 如何进入 CNS、鞘内 B 细胞如何 细胞促进 NMO 病变的发生，以及 AQP4-IgG 的离散亚群如何促进 CNS 受伤。我们的研究成果将消除该领域进展的障碍，并推进方法 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。