Study on virus particle (s) in the blood of patients with hepatitis C : an evaluation of its pleomorphism

在本研究中，我们对丙型肝炎患者血液中的丙型肝炎病毒颗粒进行了搜索，并通过电子显微镜观察发现了大小和形状具有多形性的病毒样颗粒。首先，我们尝试使用针对HCVE1蛋白的特异性抗体，通过间接免疫金电子显微镜显示丙型肝炎病毒颗粒，并成功地在富含丙型肝炎病毒RNA的样本中识别出丙型肝炎病毒粒子。丙型肝炎病毒样颗粒是一种55-65 nm的球形颗粒，表面突起长达6 nm，免疫学上区别于多形性病毒样颗粒。随后，我们用抗HCVE2/NS1和/或抗HCVE1抗体通过免疫金电子显微镜检测了HCVE2/NS1和HCVE1蛋白在丙型肝炎病毒颗粒表面的抗原性的一致性。这一结果表明，根据丙型肝炎病毒的基因组结构推测，HCVE1和HCVE2/NS1蛋白都是丙型肝炎病毒被膜的成分。我们的研究还表明，在丙型肝炎患者和发病率较高的非乙非丙型肝炎患者的血液中存在不同于丙型肝炎病毒的多形性病毒样颗粒。循环中多形性病毒样颗粒的流行与ALT水平的升高显著相关。虽然在任何含有这些病毒样颗粒的样本中都没有检测到GBV-C-RNA，而且基因组学特征还没有进一步实现，但应该澄清这些多形性病毒样颗粒的性质。

In this study, we searched for HCV particle in the blood of patients with hepatitis C and found virus-like particles with pleomorphism in size and shape by the electron microscopic observation. First, we attempted to visualize HCV particle by indirect immunogold electron microscopy, using specific antibodies to the HCVE1 protein, and succeeded in identifying HCV virion in HCV-RNA-rich samples. HCV virious, immunologically discriminated from pleomorphic virus-like particles, were 55-65 nm spherical particles with delicate 6 nm-long surface projections. Subsequently, we detected the coincidence of antigenic activities of HCVE2/NS1 and HCVE1 proteins on the surface of HCV particle by the immunogold electron microscopy using anti-HCVE2/NS1 and/or anti-HCVE1 anti-bodies. This result indicates that both HCVE1 and HCVE2/NS1 proteins are components of the envelope of HCV virion, as presumed on the basis of the construct of HCV genome.Our research also showed that pleomorphic virus-like particles, distinct from HCV,existed in the blood from hepatitis C patients and non-B,non-C hepatitis patients with high incidence. Prevalence of circulating pleomorphic virus-like particles was significantly correlated with the elevation in ALT levels. Although GBV-C-RNA was not detected in any samples containing these virus-like particles and genomic characterization has not been further achieved, it should be done to clarify the nature of these pleomorphic virus-like particles.