STUDY OF ANTINEURONAL ANTIBODIES

抗神经元抗体的研究

• 批准号: 3404490
• 负责人: PATRICIA M MOORE
• 金额: $ 13.4万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-09-01 至 1987-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3404490
• 关键词: autoantibody; autoimmune disorder; enzyme linked immunosorbent assay; immune complex; immunofluorescence technique; monoclonal antibody; neuroblastoma; neurons; neurophysiology; systemic lupus erythematosus; tissue /cell culture

This study evaluates spontaneously occurring antineuronal antibodies in an autoimmune mouse model of systemic lupus erythematosus. Systemic lupus erythematosus (SLE) is a multisystem disease characterized by the spontaneous production of multiple autoantibodies. Neurologic abnormalities are common in SLE but the pathogeneses are not well understood. Despite evidence that antibodies reactive with neuronal tissue are present in the serum and cerebrospinal fluid of patients with CNS-SLE, the long course of the disease and lack of access to central nervous system tissue makes human investigation difficult. (NZBxW)F1 hybrid mice spontaneously develop polyclonal B cell hyperactivity with formation of many autoantibodies including anti-DNA antibodies. Circulating immune complexes occur and immune complex deposition in the choroid plexus is described. In order to ascribe a role for antibodies in the pathogenesis of central nervous system disease, the following criteria should be met: 1. determine whether antibodies reactive with neurons exist, 2. demonstrate access of antibodies to CNS, 3. describe effects of antibody on in vitro neuronal function, 4. determine if antibody can produce dysfunction in vivo. In my preliminary studies antineuronal antibodies have been identified in the serum by indirect immunofluorescence against frozen brain slices and cultured neuroblastoma cells and by an ELISA assay using primary brain cultures. Documenting that these antibodies are present within the CNS is being pursued by immunoelectrophoresis of bound and unbound proteins from brain slices. The relevance of these antibodies will depend on whether or not they affect neuronal function. In order to study the antineuronal antibodies in more detail: 1. monoclonal antibodies will be made and selected for reactivity on primary brain cultures, 2. experiments are planned to determine the localization of binding of monoclonal antibody to specific cell populations within the CNS (antigenic specificity is also being studied by immunoblot and immunoprecipitation techniques), 3. in vitro effects on neuronal function will be investigated in terms of effect of antibody on a. spontaneous activity, b. repetitive firing of transcellular recordings of cultured neurons and c. resting membrane potential. 4. in vivo studies are planned in which labeled monoclonal antibodies will be administered to mice.

这项研究评估了自发产生的抗神经元抗体。 系统性红斑狼疮自身免疫性小鼠模型的建立。系统性红斑狼疮 红斑狼疮(SLE)是一种多系统疾病，以 自发产生多种自身抗体。神经学 异常在系统性红斑狼疮中很常见，但发病机制尚不清楚 明白了。尽管有证据表明抗体与神经元组织发生反应 存在于CNS-SLE患者的血清和脑脊液中， 该病病程较长，无法接触到中枢神经系统 组织使人类的研究变得困难。(NZBxW)F1杂交鼠 自发性地发生多克隆B细胞过度活动并形成 许多自身抗体，包括抗DNA抗体。循环免疫 在脉络丛中出现了复合体和免疫复合体沉积 描述。为了确定抗体在发病机制中的作用 对于中枢神经系统疾病，应满足以下标准： 1.确定是否存在与神经元反应的抗体；2.证明 抗体对中枢神经系统的访问，3.描述抗体对体外实验的影响 神经功能，4.确定抗体是否会在体内造成功能障碍。 在我的初步研究中，发现了抗神经细胞抗体 冰冻脑片间接免疫荧光法检测血清 原代培养神经母细胞瘤细胞及其酶联免疫吸附试验 文化。证明这些抗体存在于中枢神经系统是 结合蛋白和非结合蛋白的免疫电泳法 脑片。这些抗体的相关性将取决于是否或 而不是影响神经功能。为了研究抗神经细胞 更详细的抗体：1.将制备单抗和 选择在原代脑培养上的反应性，2.实验是 计划确定与单抗结合的定位 中枢神经系统内的特定细胞群(抗原特异性也是 正在用免疫印迹和免疫沉淀技术研究)，3.在 体外对神经元功能的影响将从效应的角度进行研究。 抗体对A.自发活动，B.重复激发 培养神经元和C.静息膜的跨细胞记录 潜力。4.计划进行体内研究，其中标记的单抗 抗体将被注射到老鼠身上。