A study on autoimmune mechanism in the pathogenesis of myocarditis of cardiomyopathy analysis by anti-heart monoclonal antibody

抗心脏单克隆抗体分析心肌病心肌炎发病机制中的自身免疫机制

• 批准号: 02454255
• 负责人: MATSUMORI Akira
• 金额: $ 3.39万
• 依托单位: Kyoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1990
• 资助国家: 日本
• 起止时间: 1990 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-02454255/
• 关键词: Cardiomyopathy; Myocarditis; Virus; Autoantibody; Anti-heart antibody; Auto-immunity; Monoclonal antibody; Hybridoma; 心筋炎; ハイブリド-マ

In order to clarify the role of antiheart antibody in the pathogenesis of myocarditis and cardiomyopathy, we analyzed anti-heart antibody by developing monocional antibodies. Anti-heart autoantibodies were found in the sera from Balb/c mice with myocarditis infected with encephalomyocarditis (EMC) virus. Frozen sections of the hearts of uninfected mice were incubated with mouse sera and stained with anti-mouse lgG and lgM. Positive granular staining was first seen on day 4 and persisted to day 90 ; the titer was highest on day 21. Spleen cells from mice with positive anti-heart antibody on day 21 at post-infection were fused with a cell line of plasmacytoma. Supernatants of hybridoma were screened by immunoflurerescence against murine heart sections. Two clones of lgM-producing hybridoma were developed ; one clone IA5, stained myocytes with striated pattern and the other clone IB5, stained endothelial cells. Results from western immunoblotting analyses demonstrated that IA5 reacted with cardiac myosin. These studies suggest that myosin is one of the major autoantigens in EMC virus myocarditis.

为了阐明抗心脏抗体在心肌炎和心肌病发病机制中的作用，我们通过制备抗心脏抗体，对其进行了分析。本文报道了用脑心肌炎病毒（EMC）感染Balb/c小鼠，在其心肌炎血清中检测到抗心脏自身抗体。将未感染小鼠的心脏的冷冻切片与小鼠血清一起孵育并用抗小鼠IgG和IgM染色。第4天首次观察到阳性颗粒染色，并持续至第90天;第21天滴度最高。将感染后第21天抗心脏抗体阳性的小鼠脾细胞与浆细胞瘤细胞系融合。用小鼠心脏切片免疫荧光法筛选杂交瘤上清。产生IgM的杂交瘤的两个克隆被开发;一个克隆IA 5，染色具有条纹图案的肌细胞，另一个克隆IB 5，染色内皮细胞。Western免疫印迹分析结果表明，IA 5与心肌肌球蛋白反应。提示肌球蛋白是EMC病毒性心肌炎的主要自身抗原之一。

项目成果

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Matsumori A: "Mechanism and significance of myocardial uptake of antimyosin antibody in myocarditis and cardiomyopathy:Clinical and experimental studies." SpringerーVerlag,

Matsumori A：“心肌炎和心肌病中心肌摄取抗肌球蛋白抗体的机制和意义：临床和实验研究。”

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Matsumori A: "Pathogenesis of myocardial injury in myocarditis and cardiomyopathy." Jpn Circ J.

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