Extrahepatic and Intrahepatic Replication and Expression of Hepatitis C Virus

HepatitisCvirus(HCV)isthemajorpathogenofpost--transfusionalhepatitiswithachroniccourseinupto60%ofindividuals,leadingtocirrhosisandeventuallytohepatocellularcarcinomain20%ofindividuals1'2].HCVcontainsasingleplusstrandRNAwhichissimilartopestivirusandflavivirusingenomestructures.HCVRNAcanbedetectedinsera,livertissues,peripheralbloodmononuclearcells(PBMCs)andsalivaglandsbypolymerasechainreaction(PCR)assay'-'j.Inthisstudy,theinsituhybridizationandimmunohistochemicaltechniqueusingdigoxinla…

Extrahepatic and intrahepatic replication and expression of hepatitis C virus

In order to determine the replication sites of hepatitis C virus, the in situ hybridization and immunohistochemical technique using digoxin-labeled 531bp plus-strand and minus-strand HCVRNA probes were employed to detect HCVRNA in the liver tissues, bone marrow mononuclear cells and peripheral blood mononuclear cells (PBMCs) from the patients with chronic hepatitis C, and in HCV transfected COS cells. The results showed that both plus-strand and minusstrand HCVRNA were detected in 80% of liver tissues (4/5). Plus-strand HCVRNA could be detected in 90% of PBMCs and bone marrow mononuclear cells (18/20), minus-strand HCVRNA in 25% of PBMCs. In HCV transfected COS cells, plus-strand HCVRNA distributed evenly in 20% cellular nuclei and cytoplasms. No minus-strand HCVRNA was detected in the bone marrow mononuclear cells and HCV transfected COS cells. The positive signal appeared in more cells when the liver tissues, PBMCs and marrow mononuclear cells were hybridized by plus-strand probes than when hybridized by minus-strand probes. Our results suggested that the hepatocytic cytoplasms and PBMC cytoplasms were the replication sites of HCV, but the marrow mononuclear cells were not the replication sites of HCV although they were infected by HCV. HCV infection might be accounted for the pathogenesis of chronic hepatitis and relapse of hepatitis C after liver transplantation.