Fukuda Taku, Motomura Masakatsu, Nakao Yoko, Shiraishi Hirokazu, Yoshimura Toshiro, Iwanaga Keisuke, Tsujihata Mitsuhiro, Eguchi Katsumi
First Department of Internal Medicine, Nagasaki University School of Medicine, Japan.
Ann Neurol. 2003 Jan;53(1):21-8. doi: 10.1002/ana.10392.
The aim of this study was to clarify whether autoimmunity against P/Q-type voltage-gated calcium channels (VGCCs) in the cerebellum was associated with the pathogenesis of paraneoplastic cerebellar degeneration (PCD) with Lambert-Eaton myasthenic syndrome (LEMS). We used human autopsy cerebellar tissues from three PCD-LEMS patients and six other disease patients including one with LEMS as the controls. We compared cerebellar P/Q-type VGCC in these patients and controls for the amount and ratio of autoantibody-channel complex using an 125I-omega-conotoxin MVIIC-binding assay with Scatchard analysis, and their distribution using autoradiography. The quantity of cerebellar P/Q-type VGCC measured by Scatchard analysis were reduced in PCD-LEMS patients (63.0 +/- 7.0 fmol/mg, n = 3), compared with the controls (297.8 +/- 38.9 fmol/mg, n = 6). The ratio of autoantibody-VGCC complexes to total P/Q-type VGCCs measured by immunoprecipitation assay were increased in PCD-LEMS patients. We analysed cerebellar specimens by autoradiography using (125)I-omega-conotoxin MVIIC, which specifically binds to P/Q-type VGCCs. In PCD-LEMS cerebellum, the toxin binding sites of P/Q-type VGCCs were markedly reduced compared with controls, especially in the molecular layer, which is the richest area of P/Q-type VGCCs in the normal cerebellum. This suggests that P/Q-type VGCCs of the cerebellar molecular layer is the immunological target in developing PCD-LEMS.
本研究的目的是阐明小脑针对P/Q型电压门控钙通道(VGCCs)的自身免疫是否与伴兰伯特-伊顿肌无力综合征(LEMS)的副肿瘤性小脑变性(PCD)的发病机制相关。我们使用了来自3例PCD-LEMS患者和6例其他疾病患者(包括1例LEMS患者作为对照)的人类尸检小脑组织。我们使用125I-ω-芋螺毒素MVIIC结合试验及Scatchard分析,比较了这些患者和对照中小脑P/Q型VGCC的自身抗体-通道复合物的量和比例,并使用放射自显影术比较了它们的分布。通过Scatchard分析测得,PCD-LEMS患者(63.0±7.0 fmol/mg,n = 3)小脑P/Q型VGCC的量低于对照(297.8±38.9 fmol/mg,n = 6)。通过免疫沉淀试验测得,PCD-LEMS患者中自身抗体-VGCC复合物与总P/Q型VGCC的比例增加。我们使用特异性结合P/Q型VGCC的(125)I-ω-芋螺毒素MVIIC通过放射自显影术分析小脑标本。在PCD-LEMS患者的小脑中,与对照相比,P/Q型VGCC的毒素结合位点明显减少,尤其是在分子层,而分子层是正常小脑中P/Q型VGCC最丰富的区域。这表明小脑分子层的P/Q型VGCC是PCD-LEMS发病过程中的免疫靶点。