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Detection of enteroviral genome in the myocardium from patients with viral myocarditis and dilated cardiomyopathy, and animals with Coxsackievirus myocarditis

病毒性心肌炎和扩张型心肌病患者以及柯萨奇病毒性心肌炎动物心肌中肠道病毒基因组的检测

基本信息

批准号：
07670818
负责人：
DEGUCHI Hirofumi
金额：
$ 1.41万
依托单位：
Osaka Medical College
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1996
项目状态：
已结题

项目摘要

To study the viral etiology of dilated cardiomyopathy (DCM), we examined for the presence of enteroviral RNA in myocardial biopsics from patients with DCM,myocarditis (MC), or other cardiac discases, and myocardial fissues of animal models of coxasckievirus (CV) B3 myocarditis, using polymerase chain reaction (PCR) and in situ hybridization (ISH).Myocardial biopsies from 154 patients were examined for the presence of enteroviral genome in NC,NS and POL region by RT-PCR.The positive incidence was 20% and there was no significant difference among DCM,MC and other cardiac diseases. We also examined the genotypes of enteroviruses detected in myocardial biopsies by PCR-SSCP analysis. Most of the genotypes were not corresponded with those of CV group B and a particular type of enterovirus was not present in DCM heart. In animal models of CVB3 virus myocarditis, the viral genome was detected up to the 28th day of inoculation in C3H/He mice. Light microscopic ISH with a cDNA probe for CVB showed clusters of positive signals in the areas of myocardial necrosis and cell infiltration. With electron microscopic ISH,CVB3-RNA was detected in the cytoplasm of cardiocytes, between the myofibrils, near the mitochondria, and in tubular or vesicular structures. Viral RNA was also detected in necrotic debri, in the cytoplasm of macrophages and fibroblasts. These findings suggest that CVB3 RNA is replicated in the cytoplasm of cardiocytes, transferred into tubular or vesicular structures, released into the interstitium, and phagocytosed by macrophages. Some positive signals were also detected in the cytoplasm of cardiocytes showing close contact with infiltrating lymphocytes, suggesting that the lymphocytes recognized virus-infected cardiocytes and caused to cell-mediated immune cardiocyte damage.

为了研究扩张型心肌病（DCM）的病毒病因，我们检测了DCM、心肌炎（MC）或其他心脏疾病患者的心肌活检以及柯萨奇病毒（CV）B3心肌炎动物模型的心肌破裂中肠道病毒RNA的存在，使用聚合酶链反应（PCR）和原位杂交（ISH）采用RT-PCR方法检测154例心肌活检标本NC、NS和POL区肠道病毒基因组，阳性率为20%，在DCM、MC和其他心脏疾病之间无显著性差异。我们还通过PCR-SSCP分析检测了心肌活检中检出的肠道病毒的基因型。大部分基因型与CV组B型不一致，DCM心肌中不存在一种特殊的肠道病毒。在CVB 3病毒性心肌炎动物模型中，在C3 H/He小鼠中检测到病毒基因组，直至接种的第28天。CVB cDNA探针原位杂交显示心肌坏死和细胞浸润区域有成簇的阳性信号。电镜原位杂交结果显示，心肌细胞胞质内、肌原纤维间、线粒体附近、管状或囊状结构中均可见CVB 3-RNA。在坏死碎屑、巨噬细胞和成纤维细胞的细胞质中也检测到病毒RNA。这些发现表明，CVB 3 RNA在心肌细胞的细胞质中复制，转移到管状或囊泡结构中，释放到肺泡中，并被巨噬细胞吞噬。心肌细胞胞浆中也检测到一些阳性信号，与浸润的淋巴细胞紧密接触，提示淋巴细胞识别病毒感染的心肌细胞，引起细胞介导的免疫性心肌细胞损伤。