Division of Molecular Neuroimmunology, Department of Neurology, University of Heidelberg, Heidelberg, Germany.
J Neuroinflammation. 2010 Mar 12;7:21. doi: 10.1186/1742-2094-7-21.
We report on a newly discovered serum and cerebrospinal fluid (CSF) reactivity to Purkinje cells (PCs) associated with subacute inflammatory cerebellar ataxia. The patient, a previously healthy 33-year-old lady, presented with severe limb and gait ataxia, dysarthria, and diplopia two weeks after she had recovered from a common cold. Immunohistochemical studies on mouse, rat, and monkey brain sections revealed binding of a high-titer (up to 1:10,000) IgG antibody to the cerebellar molecular layer, Purkinje cell (PC) layer, and white matter. The antibody is highly specific for PCs and binds to the cytoplasm as well as to the inner side of the membrane of PC somata, dendrites and axons. It is produced by B cell clones within the CNS, belongs to the IgG1 subclass, and activates complement in vitro. Western blotting of primate cerebellum extract revealed binding of CSF and serum IgG to an 80-97 kDa protein. Extensive control studies were performed to rule out a broad panel of previously described paraneoplastic and non-paraneoplastic antibodies known to be associated with cerebellar ataxia. Screening of >9000 human full length proteins by means of a protein array and additional confirmatory experiments revealed Rho GTPase activating protein 26 (ARHGAP26, GRAF, oligophrenin-1-like protein) as the target antigen. Preadsorption of the patient's serum with human ARHGAP26 but not preadsorption with other proteins resulted in complete loss of PC staining. Our findings suggest a role of autoimmunity against ARHGAP26 in the pathogenesis of subacute inflammatory cerebellar ataxia, and extend the panel of diagnostic markers for this devastating disease.
我们报告了一种新发现的与亚急性炎症性小脑共济失调相关的血清和脑脊液(CSF)对浦肯野细胞(PC)的反应。患者为 33 岁健康女性,在普通感冒痊愈后两周出现严重的肢体和步态共济失调、构音障碍和复视。对小鼠、大鼠和猴脑切片的免疫组织化学研究显示,高滴度(高达 1:10,000)IgG 抗体与小脑分子层、浦肯野细胞(PC)层和白质结合。该抗体高度特异性地针对 PC,并结合 PC 体细胞、树突和轴突的细胞质以及膜内侧面。它由中枢神经系统内的 B 细胞克隆产生,属于 IgG1 亚类,并在体外激活补体。灵长类动物小脑提取物的 Western blot 显示 CSF 和血清 IgG 与 80-97 kDa 蛋白结合。进行了广泛的对照研究以排除广泛的先前描述的与小脑共济失调相关的副肿瘤和非副肿瘤抗体。通过蛋白质阵列对 >9000 个人全长蛋白质进行筛选,并进行额外的确认实验,发现 Rho GTPase 激活蛋白 26(ARHGAP26、GRAF、寡肽-1 样蛋白)是靶抗原。用人类 ARHGAP26 预先吸附患者的血清而不是用其他蛋白质预先吸附导致 PC 染色完全丢失。我们的研究结果表明,针对 ARHGAP26 的自身免疫在亚急性炎症性小脑共济失调的发病机制中起作用,并扩展了该毁灭性疾病的诊断标志物面板。