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

本研究在丙型肝炎患者血液中寻找丙型肝炎病毒颗粒，通过电镜观察发现病毒样颗粒大小和形状具有多态性。首先，我们尝试通过间接免疫金电子显微镜观察HCV颗粒，使用针对HCVE 1蛋白的特异性抗体，并成功地在HCV-RNA丰富的样品中鉴定出HCV病毒体。HCV病毒，从免疫学上区别于多形性病毒样颗粒，是55-65 nm的球形颗粒，具有6 nm长的精细表面突起。用抗HCVE 2/NS 1和/或抗HCVE 1抗体，通过免疫金电镜技术检测HCV颗粒表面HCVE 2/NS 1和HCVE 1蛋白的抗原活性一致性。这一结果表明，HCVE 1和HCVE 2/NS 1蛋白均为HCV包膜蛋白，与HCV基因组的结构一致，并发现丙型肝炎患者和非乙、非丙型肝炎患者血液中存在与HCV不同的多形病毒样颗粒。循环多形性病毒样颗粒的患病率与ALT水平的升高显著相关。尽管在含有这些病毒样颗粒的任何样品中均未检测到GBV-C-RNA，并且尚未进一步实现基因组表征，但应进行研究以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.