| Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA |
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| 作者姓名： | Yao YQ Zhang DF Tang N Huang AL Zou XY Xiao JF Luo Y Zhang DZ Wang B Zhou WP Ren H Liu Q Guo SH |
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| 作者单位： | Yun-Qing Yao，Xiao-Yi Zou，Jiang-Feng Xiao，Department of Infectious Diseases of the First Affiliated Hospital，Chongqing University of Medical Sciences，Chongqing 400016，China Ding-Feng Zhang，Ni Tang，Ai-Long Huang，Yun Luo，Da-Zhi Zhang，Bo Wang，Wei-Ping Zhou，Hong Ren，Qi Liu，Shu-Hua Guo，Institute for Viral Hepatitis，Chongqing University of Medical Sciences，Chongqing 400010，China |
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| 基金项目： | Supported by the National Natural Science Foundation of China,No. 39670340
the Applied Basic Research Programs of Science
Technology Commission Foundation of Chongqing, No.20021889 |
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| 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, 1.19×1012 copies of linear HBV DNA/1×107 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.
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| 关 键 词： | 乙型肝炎病毒 病毒复制 原发性肝细胞损伤 病毒转染 线形滤过性病毒 病毒DNA |
| 收稿时间： | 2004 Nov 2 |
Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA |
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| Yao YQ,Zhang DF,Tang N,Huang AL,Zou XY,Xiao JF,Luo Y,Zhang DZ,Wang B,Zhou WP,Ren H,Liu Q,Guo SH.Replication of hepatitis B virus in primary duck hepatocytes transfected with linear viral DNA[J].World Journal of Gastroenterology,2005,11(32):5019-5021. |
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| Authors: | Yao Yun-Qing Zhang Ding-Feng Tang Ni Huang Ai-Long Zou Xiao-Yi Xiao Jiang-Feng Luo Yun Zhang Da-Zhi Wang Bo Zhou Wei-Ping Ren Hong Liu Qi Guo Shu-Hua |
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| Institution: | 1. Department of Infectious Diseases of the First Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400016, China
2. Institute for Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400010, China |
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| Abstract: | 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, 1.19 x 10(12) copies of linear HBV DNA/1 x 10(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 > or =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. |
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| Keywords: | Hepatitis B virus Replication Expression Primary duck hepatocytes |
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