An Immunologic Approach Alter Function of Neuronal Genes

改变神经元基因功能的免疫学方法

• 批准号: 6333822
• 负责人: MATTHEW J DURING
• 金额: $ 31.4万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6333822
• 关键词: NMDA receptors; active immunization; antibody formation; antireceptor antibody; autoantibody; electrophysiology; epilepsy; genetically modified animals; laboratory mouse; passive immunization; vector vaccine; voltage /patch clamp

DESCRIPTION (From the Applicant's Abstract): The brain is generally considered immunoprivileged, although increasing examples of immunological responses to brain antigens, neuronal expression of Major Histocompatibility Class I genes and neurological autoimmunity have been recognized. In this application we propose the use of vaccination strategies to generate autoantibodies which target a specific brain protein, the NR1 subunit of the N-methyl-D-aspartate (NMDA) receptor. We present preliminary data to show the development of a novel vaccine, the peroral administration of AAV vectors targeting the lamina propria resulting in stable transgene expression in a variety of cells including professional antigen presenting cells (APC). This viral vaccine is associated with a humoral response to the transgene, both foreign and self-antigens, resulting in circulating autoantibodies in this latter case. Moreover, we show that the administration of NMDAR1 -expressing vectors, but not control vectors, is associated with anti-epileptic and neuroprotective activity in experimental animal models. In this grant application we propose to extend these preliminary studies to further define the mechanism of action of this self-antigen immunization and to determine whether such a vaccination strategy targeting brain proteins is feasible and may have therapeutic potential for neurological disorders. The proposal is now focused on testing the hypothesis that the neuroprotective phenotype we observe is mediated via antibodies. Secondly, we will explore the mechanism of the vaccination, see if the observation is generalizable to other cell surface receptors, and also compare this approach with other more traditional protein vaccination strategies. It includes the use of adoptive transfer methods to definitively test the hypothesis of an antibody mediated mechanism as well as the use of CD4 and CD8 knockout mice to help dissect out which components of the immune system mediated the phenotypic effects. It characterizes and further defines the mechanism of action of the vaccination using electrophysiological and calcium imaging studies on primary neurons and it also involves testing a variety of approaches in an established model of neurological disease. The successful completion of these experiments is likely to represent a significant advance in the development of a novel therapeutic approach for neurological disorders.

描述（来自申请人的摘要）：大脑通常被认为 免疫豁免，尽管免疫反应的例子越来越多 脑抗原、主要组织相容性 I 类基因的神经元表达 和神经系统自身免疫已得到认可。在这个应用程序中我们 提议使用疫苗接种策略来产生自身抗体， 靶向特定的脑蛋白，即 N-甲基-D-天冬氨酸的 NR1 亚基 (NMDA) 受体。我们提供初步数据来展示小说的发展 疫苗，口服 AAV 载体靶向固有层 导致多种细胞中稳定的转基因表达，包括 专职抗原呈递细胞（APC）。这种病毒疫苗与 对转基因（外来抗原和自身抗原）产生体液反应， 在后一种情况下导致循环自身抗体。此外，我们还展示了 施用表达NMDAR1的载体，但不施用对照载体， 与实验中的抗癫痫和神经保护活性有关 动物模型。在此拨款申请中，我们建议延长这些初步 研究进一步明确这种自身抗原的作用机制 免疫并确定这种疫苗接种策略是否针对 脑蛋白是可行的，并且可能具有神经系统疾病的治疗潜力 失调。该提案现在的重点是检验以下假设： 我们观察到的神经保护表型是通过抗体介导的。其次，我们 将探索疫苗接种的机制，看看观察结果是否正确 可推广到其他细胞表面受体，并比较这种方法 与其他更传统的蛋白质疫苗接种策略。它包括使用 过继转移方法来明确检验抗体的假设 介导的机制以及使用 CD4 和 CD8 敲除小鼠来帮助 剖析免疫系统的哪些成分介导表型 影响。它表征并进一步定义了作用机制 使用电生理学和钙成像研究对初级儿童进行疫苗接种 神经元，它还涉及在已建立的环境中测试各种方法 神经系统疾病模型。这些实验的顺利完成 可能代表小说发展的重大进步 神经系统疾病的治疗方法。