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: 9198012
• 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: 2013
• 资助国家: 美国
• 起止时间: 2013-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9198012
• 关键词: Address; Affect; Antibodies; Antibody Repertoire; Antibody Response; Antigen Targeting; Antigens; Area; Astrocytes; Autoantibodies; Autoimmune Responses; Automobile Driving; Binding; Biological Assay; Biological Models; Blindness; Blood - brain barrier anatomy; Brain; Brain Injuries; Cell Line; Cell-Mediated Cytolysis; Cells; Centers for Disease Control and Prevention (U.S.); Clinical; Complement; Complement Activation; Demyelinating Diseases; Demyelinations; Development; Diagnosis; Disease; Disease model; Early Diagnosis; Epitopes; Equipment and supply inventories; Event; Fluorescence-Activated Cell Sorting; Foot Process; Frequencies; Human; Humoral Immunities; Immune Targeting; Immune response; Immune system; Immunoglobulin G; In Vitro; Individual; Inflammatory; Injection of therapeutic agent; Injury; Lesion; Leukocytes; Libraries; Link; Measures; Mediating; Mediator of activation protein; Metabolic; Metabolism; Modeling; Mus; Myelin; Nature; Neuraxis; Neuroglia; Neurologic; Neuromyelitis Optica; Neuronal Injury; Neurotransmitters; Oligodendroglia; Optic Nerve; Paralysed; Pathogenesis; Pathogenicity; Pathology; Patient Care; Patients; Pattern; Physicians; Plasma Cells; Plasmablast; Population; Reagent; Recombinant Antibody; Recombinants; Recruitment Activity; Relapse; Resources; Reverse Transcriptase Polymerase Chain Reaction; Risk; Role; Series; Serum; Severities; Severity of illness; Specificity; Spinal Cord; System; Technology; Tissues; Transgenic Animals; antibody-dependent cell cytotoxicity; aquaporin 4; aquaporin-2; central nervous system demyelinating disorder; central nervous system injury; cytokine; disability; disease diagnosis; excitotoxicity; experimental study; high risk; human monoclonal antibodies; immunopathology; in vivo; in vivo Model; inhibitor/antagonist; innovation; kainate; novel; novel therapeutics; public health relevance; response; seropositive; skills; water channel; young adult

DESCRIPTION (provided by applicant): Neuromyelitis optica (NMO) is a severe human demyelinating disorder with predilection for the optic nerve and spinal cord. Approximately 70% of affected individuals demonstrate a serum autoantibody response, NMO-IgG, against the aquaporin-4 (AQP4) water channel. NMO-IgG binding to AQP4 in astrocytes in the central nervous system (CNS) is thought to initiate a series of inflammatory events, including complement- and cell-mediated astrocyte damage, leukocyte recruitment, cytokine release, and myelin damage. Using single- cell FACS and recombinant antibody technology, we have faithfully reconstructed the intrathecal humoral immune response in NMO and demonstrated that AQP4 autoantibodies are sufficient to initiate astrocyte injury and disease pathology. To further the care of patients with human demyelinating disease, we will use our novel resources and innovative approach to address fundamental gaps in our understanding of NMO pathogenesis: 1) Are there alternative targets of the immune response in NMO? 2) Does the nature of the AQP4 immune response correlate with disease severity? 3) What are the mechanisms driving astrocyte injury and myelin damage In NMO pathology? First, to identify novel targets of the autoimmune response in NMO, we will examine non-AQP4 specific NMO recombinant antibodies (rAbs) for binding to human and murine tissue, glial cell lines and primary glial cultures. A compendium of candidate rAbs will be generated from our existing antibody repertoires as well as new cases of AQP4 seronegative disease. Second, to investigate the association between NMO-IgG specificity and clinical disease activity, we will evaluate the relationship between two distinct patterns of AQP4 epitope specificity and the frequency and severity of clinical activity. We hypothesize that the spectrum of AQP4 epitope specificity in NMO-IgG will influence antibody effector function and CNS injury. And third, we will define the mechanisms underlying AQP4-mediated astrocyte injury and myelin damage using a combination of disease models, transgenic animals, and NMO CSF rAbs. Specifically, we will use mixed glial cultures, ex vivo spinal cord explants, and intracerebral injections to evaluate th effect of complement activation, antibody-dependent cell mediated cytotoxicity, excitotoxicity, cytokine release, and astrocyte metabolism on AQP4 antibody-mediated CNS injury. The detailed characterization of the humoral immune response in NMO will aid in the diagnosis and treatment of NMO, the identification of novel target antigens in human demyelinating disease, and the elucidation of the mechanisms causing glial and neuronal injury in CNS inflammatory disease.

描述（由申请人提供）：视神经脊髓炎（NMO）是一种严重的人类脱髓鞘疾病，好发于视神经和脊髓。大约 70% 的受影响个体表现出针对水通道蛋白 4 (AQP4) 水通道的血清自身抗体反应 (NMO-IgG)。 NMO-IgG 与中枢神经系统 (CNS) 星形胶质细胞中的 AQP4 结合被认为会引发一系列炎症事件，包括补体和细胞介导的星形胶质细胞损伤、白细胞募集、细胞因子释放和髓磷脂损伤。利用单细胞FACS和重组抗体技术，我们忠实地重建了NMO中的鞘内体液免疫反应，并证明AQP4自身抗体足以引发星形胶质细胞损伤和疾病病理。为了进一步护理人类脱髓鞘疾病患者，我们将利用我们的新资源和创新方法来解决我们对 NMO 发病机制理解中的根本差距：1) NMO 中的免疫反应是否存在替代目标？ 2) AQP4 免疫反应的性质与疾病严重程度相关吗？ 3）NMO病理中星形胶质细胞损伤和髓磷脂损伤的驱动机制是什么？ 首先，为了确定 NMO 中自身免疫反应的新靶标，我们将检查非 AQP4 特异性 NMO 重组抗体 (rAb) 与人和小鼠组织、神经胶质细胞系和原代神经胶质培养物的结合。候选 rAb 的概要将从我们现有的抗体库以及 AQP4 血清阴性疾病的新病例中生成。其次，为了研究 NMO-IgG 特异性与临床疾病活动之间的关联，我们将评估 AQP4 表位特异性的两种不同模式与临床活动的频率和严重程度之间的关系。我们假设 NMO-IgG 中 AQP4 表位特异性的谱将影响抗体效应器功能和 CNS 损伤。第三，我们将 使用疾病模型、转基因动物和 NMO CSF rAb 的组合来定义 AQP4 介导的星形胶质细胞损伤和髓磷脂损伤的机制。具体来说，我们将使用混合神经胶质培养物、离体脊髓外植体和脑内注射来评估补体激活、抗体依赖性细胞介导的细胞毒性、兴奋性毒性、细胞因子释放和星形胶质细胞代谢对 AQP4 抗体介导的 CNS 损伤的影响。 NMO 体液免疫反应的详细表征将有助于 NMO 的诊断和治疗、人类脱髓鞘疾病中新靶抗原的鉴定以及中枢神经系统炎症性疾病中引起神经胶质和神经元损伤的机制的阐明。