The Autoimmune Response in Batten Disease

Batten 病的自身免疫反应

• 批准号: 6935189
• 负责人: DAVID A. PEARCE
• 金额: $ 33.99万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6935189
• 关键词: SDS polyacrylamide gel electrophoresis; antibody titering; antigen antibody reaction; autoantibody; autoimmunity; cerebellar Purkinje cell; complement; cytokine; disease /disorder model; enzyme activity; gene expression; gene mutation; gene targeting; genetically modified animals; glutamate decarboxylase; glutamate receptor; glutamate transporter; immunocytochemistry; immunoglobulin G; in situ hybridization; laboratory mouse; neuronal ceroid lipofuscinosis; organ culture; passive immunization; pathologic process

DESCRIPTION (provided by applicant): Batten, with an incidence as high as one in 12,500 live births. Batten disease results from mutation of CLN3, and is characterized pathologically by the accumulation of autofluorescent hydrophobic material in the lysosome of neurons and other cell types. However, the mechanism driving these cellular alterations and the manner in which they relate to the neurodegeneration in Batten disease is unknown. Individuals with Batten disease, and a cln3-knockout mouse model for Batten disease have a circulating autoantibody to glutamic acid decarboxylase (GAD65) that is inhibitory to this enzyme's ability to convert glutamic acid to 7-aminobutyric acid (GABA). The GAD65 autoantibody associates with the brain in cln3-knockout mice, inhibits the activity of GAD, and results in a subsequent elevation of glutamate and altered expression of genes involved in the regulation and utilization of glutamate. The major goal of this proposal is to establish and characterize the contribution of the GAD65 autoantibody to the pathogenesis of Batten disease. As the model for this study is a genetically defined I knockout mouse, we will establish whether the presence of a GAD65 autoantibody is simply an epiphenomenon, and whether it contributes directly to Batten disease. We shall: (i) construct cln3-knockout mice that are unable to mount an immune response to GAD65 by crossing to MuMT and C-alpha knockout mice, which lack the ability to generate B-cells, and CD4+ and CD8+ T-cells, respectively. This will enable us to determine whether the autoantibody contributes to the pathology of Batten disease. (ii) test by passive transfer of GAD65 autoantibodies to normal mice the degree of pathogenicity on the CNS mediated by these autoantibodies. Also, by transfer of GAD65 autoantibodies to immune deficient mice we will determine if the cln3-defect allows for preferential deposition of IgG in the brain (iii) define the inflammatory response to the presence of autoantibodies and the participation of the innate immune system in the disease process in cln3-knockout mice by examining cytokines such as IL-1 and TNF, complement proteins and microglial activation, respectively. (iv) characterize cell type specific molecular effects that elevated glutamate and exposure to the autoantibody have on primary neuronal culture from cln3-knockout mice.

描述（由申请人提供）：Batten，发病率高达 12,500 例活产儿中就有 1 例。 Batten病是由CLN3突变引起的，其病理特征是神经元和其他细胞类型的溶酶体中自发荧光疏水物质的积累。然而，驱动这些细胞改变的机制以及它们与巴顿病神经变性的关系尚不清楚。巴顿病患者和巴顿病 cln3 敲除小鼠模型具有针对谷氨酸脱羧酶 (GAD65) 的循环自身抗体，该抗体抑制该酶将谷氨酸转化为 7-氨基丁酸 (GABA) 的能力。 GAD65 自身抗体与 cln3 敲除小鼠的大脑结合，抑制 GAD 的活性，导致谷氨酸随后升高，并改变参与谷氨酸调节和利用的基因表达。该提案的主要目标是确定并表征 GAD65 自身抗体对巴顿病发病机制的贡献。由于本研究的模型是基因定义的 I 基因敲除小鼠，因此我们将确定 GAD65 自身抗体的存在是否只是一种附带现象，以及它是否直接导致巴顿病。我们应：（i）通过与 MuMT 和 C-alpha 敲除小鼠杂交，构建无法对 GAD65 产生免疫反应的 cln3 敲除小鼠，这些小鼠缺乏生成 B 细胞、CD4+ 和 CD8+ T 细胞的能力，分别。这将使我们能够确定自身抗体是否有助于巴顿病的病理学。 (ii)通过将GAD65自身抗体被动转移至正常小鼠来测试这些自身抗体介导的对CNS的致病性程度。此外，通过将 GAD65 自身抗体转移至免疫缺陷小鼠，我们将确定 cln3 缺陷是否允许 IgG 在大脑中优先沉积 (iii) 定义对自身抗体存在的炎症反应以及先天免疫系统在炎症反应中的参与。通过分别检查 IL-1 和 TNF 等细胞因子、补体蛋白和小胶质细胞激活，研究 cln3 敲除小鼠的疾病过程。 (iv) 表征谷氨酸升高和暴露于自身抗体对 cln3 敲除小鼠的原代神经元培养物产生的细胞类型特异性分子效应。