|In vitro resistance to interferon of hepatitis B virus with precore mutation|
|Wang, Yan; Wei, Lai; Jiang, Dong; Cong, Xu; Fei, Ran; Xiao, Jiang; Wang, Yu|
|关键词||Hepatitis b Virus Mutation Interferon Viral Resistance|
|刊名||WORLD JOURNAL OF GASTROENTEROLOGY|
|WOS标题词||Science & Technology|
|类目[WOS]||Gastroenterology & Hepatology|
|研究领域[WOS]||Gastroenterology & Hepatology|
AIM: Chronic hepatitis B virus (HBV) infection is predominantly treated with interferon alpha (IFN-alpha), which results in an efficient reduction of the viral load only in 20-40% of treated patients. Mutations at HBV precore prevail in different clinical status of HBV infection. The roles of precore mutation in the progression of chronic hepatitis and interferon sensitivity are still unknown. The aim of this study was to explore if there was any relationship between HBV precore mutation and sensitivity to interferon in vitro.
METHODS: HBV replication-competent recombinant constructs with different patterns of precore mutations were developed. Then the recombinants were transiently transfected into hepatoma cell line (Huh7) by calcium phosphate transfection method. With or without IFN, viral products in culture medium were collected and quantified 3 d after transfection.
RESULTS: We obtained 4 recombinant constructs by orientation-cloning 1.2-fold-overlength HBV genome into pUC18 vector via the EcoRI and Hind III and PCR mediated site-directed mutagenesis method. All the recombinants contained mutations within precore region. Huh7 cells transfected with recombinants secreted HBsAg and HBV particles into the cell culture medium, indicating that all the recombinants were replication-competent. By comparing the amount of HBV DNA in the medium, we found that HBV DNA in medium reflecting HBV replication efficiency was different in different recombinants. Recombinants containing precore mutation had fewer HBV DNAs in culture medium than wild type. This result showed that recombinants containing precore mutation had lower replication efficiency than wild type. HBV DNA was decreased in pUC18-HBV1.2-WT recombinants after IFN was added while others with precore mutations were not, indicating that HBV harboring precore mutation was less sensitive to IFN in cell culture system.
CONCLUSION: These data indicate that HBV harboring precore mutation may be resistant to IFN in vitro. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.
|资助机构||Major State Basic Research Development Program of China (973 Program)|
|作者单位||Peking Univ, Peoples Hosp, Inst Hepatol, Beijing 100044, Peoples R China|
|Wang, Yan,Wei, Lai,Jiang, Dong,et al. In vitro resistance to interferon of hepatitis B virus with precore mutation[J]. WORLD JOURNAL OF GASTROENTEROLOGY,2005,11(5):649-655.|
|APA||Wang, Yan.,Wei, Lai.,Jiang, Dong.,Cong, Xu.,Fei, Ran.,...&Wang, Yu.(2005).In vitro resistance to interferon of hepatitis B virus with precore mutation.WORLD JOURNAL OF GASTROENTEROLOGY,11(5),649-655.|
|MLA||Wang, Yan,et al."In vitro resistance to interferon of hepatitis B virus with precore mutation".WORLD JOURNAL OF GASTROENTEROLOGY 11.5(2005):649-655.|