Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA

被引：0

作者：

Yao, Yun-Qing ^{[1
]}

Zhang, Ding-Feng ^{[2
]}

Tang, Ni ^{[2
]}

Huang, Ai-Long ^{[2
]}

Zou, Xiao-Yi ^{[1
]}

Xiao, Jiang-Feng ^{[1
]}

Luo, Yun ^{[2
]}

Zhang, Da-Zhi ^{[2
]}

Wang, Bo ^{[2
]}

Zhou, Wei-Ping ^{[2
]}

Ren, Hong ^{[2
]}

Liu, Qi ^{[2
]}

Guo, Shu-Hua ^{[2
]}

机构：

[1] Chongqing Univ Med Sci, Affiliated Hosp 1, Dept Infect Dis, Chongqing 400016, Peoples R China

[2] Chongqing Univ Med Sci, Inst Viral Hepatitis, Chongqing 400010, Peoples R China

来源：

WORLD JOURNAL OF GASTROENTEROLOGY | 2005年 / 11卷 / 32期

基金：

中国国家自然科学基金;

关键词：

Hepatitis B virus; Replication; Expression; Primary duck hepatocytes;

D O I：

10.3748/wjg.v11.i32.5019

中图分类号：

R57 [消化系及腹部疾病];

学科分类号：

摘要：

AIM: To explore the expression and replication of hepatitis B virus (HBV) DNA in primary duck hepatocytes (PDHs). METHODS: Complete HBV genome was transfected into PDHs by electroporation (transfected group, 1.19x10(12) copies of linear HBV DNA/1x10(7) PDHs). After 1-5 d of transfection, HBsAg and HBeAg in the supernatant and lysate of PDHs were measured with the IMX System. Meanwhile, replicative intermediates of HBV DNA were analyzed by Southern blotting and Dot blotting. PDHs electroporated were used as control group. RESULTS: HBsAg in the hepatocyte lysates of transfected group was 15.24 (1 d), 14.55 (3 d) and 5.13 (5 d; P/N values, positive >= 2.1) respectively. HBeAg was negative (<2.1). Both HBsAg and HBeAg were negative in the supernatant of transfected group. Dot blotting revealed that HBV DNA was strongly positive in the transfected group and negative in the control group. Southern blot analysis of intracellular total DNA indicated that there were relaxed circular (rc DNA), covalently closed circular (ccc DNA), and single-stranded (ss DNA) HBV DNA replicative intermediates in the transfected group, there was no integrated HBV DNA in the cellular genome. These parameters were negative in control group. CONCLUSION: Expression and replication of HBV genes can occur in hepatocytes from non-mammalian species. HBV replication has no critical species-specificity, and yet hepatic-specific regulating factors in hepatocytes may be essential for viral replication. (C) 2005 The WJG Press and Elsevier Inc. All rights reserved.

引用

页码：5019 / 5021

页数：3

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