丙型肝炎病毒准种在单个核细胞和肝细胞内的分隔性分布

Compartmental distribution of hepatitis C virus quasispecies in mononuclear cells and liver

目的探讨丙型肝炎病毒（ＨＣＶ）对单个核细胞感染在ＨＣＶ感染慢性化中的作用。方法通过对ＨＣＶ高度变异区（ＨＶＲ）基因的体外扩增后产物的直接测序，比较慢性丙型肝炎和接受肝移植的肝硬化患者血清、外周血单个核细胞（ＰＢＭＣ）、肝脏相关的单个核细胞（ＬＡＭＣ）和肝脏中ＨＣＶ准种（ｑｕａｓｉｓｐｅｃｉｅｓ）的分布。结果（１）３例肝移植和５例慢性肝炎患者ＰＢＭＣ中的主要ＨＣＶ准种不同于肝脏中的ＨＣＶ；（２）３例患者的ＰＢＭＣ和ＬＡＭＣ中ＨＶＲ有不同的基因替代；（３）ＨＶＲ基因的核苷酸突变主要位于Ｅ２基因的前５０个氨基酸内。结论（１）单个核细胞中的ＨＣＶＲＮＡ是由于ＨＣＶ的感染所致；（２）ＨＣＶ准种在肝细胞和单个核细胞中的分隔性分布可能与ＨＣＶ感染的慢性化密切相关。

Objective To explore the role of the infection of monouclear cells by hepatitis C virus(HCV) in the chronicity of HCV infection. Methods Direct sequencing of the amplified hypervariable region (HVR) gene of HCV was used to distinguish main HCV quasispecies infecting non hepatocytes from those infecting hepatocytes and circulating virions.The main HCV quasispecies in peripheral blood mononuclear cells(PBMC), liver associated mononuclear cells (LAMC), serum and liver samples from 6 patients with chronic hepatitis C and from 6 patients with HCV related cirrhosis were analysed and compared. Results The main HCV qusispecies in the livers were different from those in PBMC in 3 transplanted cirrhotic patients and 5 patients with chronic hepatitis C. Different substitutions existed between PBMC and LAMC in 3 of 6 patients analysed. Mutations were observed within the first 50 amino acids of E2 gene,which induced amino acid change in 6 of 9 patients analysed. Conclusion In chronically infected patients, the detection of HCV RNA in mononulear cells is not due to the adsorption of circulating virions but a significant infection. The compartmental distribution of HCV quasispecies in hepatocytes and mononuclear cells could be involved in the chronicity of HCV infection.