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 视神经脊髓炎谱系障碍(NMOSD)是一种严重的自身免疫性疾病，其目标是 水通道蛋白4(AQP4)水通道。NMOSD的中枢神经系统损伤是由绑定引起的 靶向星形胶质细胞的AQP4自身抗体(AQP4-Ig G)和抗体效应功能的激活； 然而，血清AQP4-Ig G滴度与疾病复发或严重程度无关。因此，识别 影响中枢神经系统通路、鞘内生产或AQP4-Ig G效应功能的因素是 了解NMOSD的发病机制。使用单细胞分选，重组抗体技术，以及 重链可变区分析，我们重建了鞘内AQP4-Ig G 激活脑微血管内皮细胞的分离抗内皮细胞抗体对NMOSD的反应 细胞(BMEC)，并鉴定了扩增的CD27-IGD-双阴性(DN)记忆B细胞群 克隆性与鞘内AQP4特异的浆母细胞相关。我们现在做好了独特的准备来测试我们的 假设AQP4-Ig G、抗BMECs自身抗体和促炎的DNB细胞在 推动NMOSD活动和病理学的音乐会。 在目标1中，我们将检查葡萄糖调节蛋白78(GRP78)自身抗体对 NMOSD疾病活跃度。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，并测量它们增加的能力 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专一性如何 抗AQP4自身抗体影响NMOSD的病变形成和中枢神经系统损伤。我们已经确认了多个 对靶产生不同影响的患者来源的AQP4特异性重组抗体的种类 星形胶质细胞。我们将评估AQP4RAB激活补体依赖性细胞毒的能力，启动 抗体依赖细胞介导的细胞毒作用，并调节星形胶质细胞趋化因子和细胞因子的产生 体外培养。然后我们将评估这些参数如何影响动物模型和NMOSD损伤的形成 受影响患者的疾病活跃度。在目标3中，我们将分析班级转换的扩展人口 CD27-IGD-DNB细胞在NMOSD患者中的表达及其在疾病活动性中的作用富足和 我们将比较NMOSD复发和缓解期间DNB细胞的免疫球蛋白谱，我们将 评估NMOSD糖尿病肾病B细胞对不同免疫反应的细胞因子反应和免疫球蛋白产生 刺激物。此外，我们还将研究NMOSD糖尿病肾病B细胞将AQP4呈递给自体T细胞的可能性 细胞。这些研究的结果将阐明AQP4-IgG如何进入中枢神经系统，如何在鞘内B 细胞参与NMO损伤的启动，以及AQP4-Ig G的离散亚群如何对中枢神经系统起作用 受伤。我们的研究成果将消除在这一领域取得进展的障碍，并推动 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。