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目的 以常规电镜和免疫电镜技术 ,发现和证实慢性丙型肝炎患者外周血单个核细胞(PBMCs)内丙型肝炎病毒 (HCV)颗粒。试图在病毒形态学和形态发生学上证实PBMCs的HCV感染和复制。方法 以逆转录多聚酶链反应 (RT PCR)和免疫组织化学方法 ,分别检测 2 8例患者PBMCs内HCVRNA和HCVAg ,对其中阳性标本重点进行电镜研究。 结果 HCVRNA和HCVAg阳性检出率分别为 77 2 7% (17/ 2 2 )和 75 0 0 % (2 1/ 2 8) ,其中 10例HCVAg强阳性PBMCs的胞质空泡内发现直径分别约为 6 5nm和 110nm圆球形病毒颗粒 ,并可见病毒芽生形成现象。免疫电镜证实病毒颗粒及含病毒颗粒的空泡膜含HCV抗原性 ,确认所见为HCV颗粒。患者PBMCs显示核变形固缩和核 /浆比例增大等超微结构改变。结论 电镜及免疫电镜研究发现和证实了丙型肝炎患者PBMCs内复制的HCV样颗粒。从病毒形态及形态发生上证实患者PBMCs的HCV感染和复制。病毒在感染细胞胞质空泡部位合成和组装 ,以芽生方式进入胞质空泡内形成病毒颗粒。患者PBMCs显示核 /浆比例增大 ,核变形和固缩等超微结构改变。
Objective To detect and validate hepatitis C virus (HCV) particles in peripheral blood mononuclear cells (PBMCs) of patients with chronic hepatitis C by conventional electron microscopy and immunoelectron microscopy. Attempts to confirm the HCV infection and replication of PBMCs in terms of virus morphology and morphogenesis. Methods Reverse transcription polymerase chain reaction (RT PCR) and immunohistochemistry were used to detect HCV RNA and HCVAg in PBMCs of 28 patients respectively. The positive samples were examined by electron microscopy. Results The positive rates of HCV RNA and HCVAg were 77 2 7% (17/2 2) and 75 0 0% (2 1/2 8), respectively. Among the 10 HCVAg-positive PBMCs, the cytoplasmic vacuoles were found in about 10% For 6 5nm and 110nm spherical particles, and the formation of the virus budding phenomenon. Immuno-electron microscopy confirmed that the virus particles and the vacuolar membrane containing the virus particles contained HCV antigen, and confirmed that HCV particles were observed. Patient PBMCs showed ultrastructural changes such as nuclear shrinkage and nuclear / plasma ratio increase. Conclusion Electron microscopy and immunoelectron microscopy found and confirmed HCV-like particles replicated in PBMCs of hepatitis C patients. The HCV infection and replication of PBMCs in patients were confirmed from the morphological and morphological changes of virus. The virus is synthesized and assembled in the cytoplasmic vacuolar parts of the infected cells and enters the cytoplasmic vacuoles to form viral particles in a budding manner. Patients PBMCs showed nuclear / plasma ratio increases, nuclear deformation and shrinkage and other ultrastructural changes.