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）：板条，发病率高达12,500名活产。 Batten疾病是由CLN3突变引起的，并且在病理上以自动荧光疏水物材料在神经元和其他细胞类型的溶酶体中的积累进行表征。但是，驱动这些细胞改变的机制及其与棕褐色疾病中神经退行性相关的方式尚不清楚。具有板条疾病的个体和用于谷色疾病的CLN3敲除小鼠模型具有循环自身抗体对谷氨酸脱羧酶（GAD65）的循环自身抗体，该酶抑制了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敲除小鼠中的大脑相关联，抑制GAD的活性，并导致谷氨酸的升高和涉及谷氨酸调节和利用的基因表达的改变。该提案的主要目的是建立和表征GAD65自身抗体对巴顿疾病发病机理的贡献。由于这项研究的模型是一种遗传定义的I基因敲除小鼠，因此我们将确定GAD65自身抗体的存在是否仅仅是epiphenomenon，以及它是否直接促进了抗肉芽症疾病。我们将：（i）构建CLN3-敲除小鼠，该小鼠无法通过分别越过MUMT和C-Alpha敲除小鼠来安装对GAD65的免疫反应，这些小鼠缺乏产生B细胞的能力，以及CD4+和CD8+ T细胞。这将使我们能够确定自身抗体是否有助于棕褐色疾病的病理。 （ii）通过将GAD65自身抗体向正常小鼠的被动转移测试，这些自身抗体介导的中枢神经系统的致病程度。 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小胶质激活。 （iv）表征细胞类型特异性分子作用，升高谷氨酸并暴露于自身抗体对CLN3-敲除小鼠的原发性神经元培养物具有。