Viral Hepatitis
Copyright ©2007 Baishideng Publishing Group Co., Limited. All rights reserved.
World J Gastroenterol. May 7, 2007; 13(17): 2490-2495
Published online May 7, 2007. doi: 10.3748/wjg.v13.i17.2490
High expression of hepatitis B virus based vector with reporter gene in hepatitis B virus infection system
Shi-Hong Li, Wen-Ge Huang, Bing Huang, Xi-Gu Chen
Shi-Hong Li, Wen-Ge Huang, Xi-Gu Chen, Center of Experimental Animals, Sun Yat-sen University, Guangzhou 510080, Guangdong Province, China
Bing Huang, State KeyLaboratory of Ophthalmology, ZhongShan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, Guangdong Province, China
Author contributions: All authors contributed equally to the work.
Supported by the grants from the National Science Foundation of China, No. 30271177 and No. 39870676, and the Natural Science Foundation of Guangdong Province, No. 021903
Correspondence to: Xi-Gu Chen, Center of Experimental Animals, Sun Yat-sen University, 58 Zhongshan 2 Road, Guangzhou, Guangdong 510080, China.
Telephone: +86-20-87331393 Fax: +86-20-87331230
Received: March 13, 2007
Revised: March 18, 2007
Accepted: March 31, 2007
Published online: May 7, 2007

AIM: To construct a hepatitis B virus (HBV)-based vector with a reporter gene and to establish an HBV infection system to evaluate the availability of the vector.

METHODS: The HBV-based vectors with green fluorescence protein (GFP) were packaged into the liver of immunodeficient mice through transfer and helper plasmid using hydrodynamic technology. Wild type HBV (wt HBV) was provided by plasmid MC2009. Primary human hepatocytes (PHH) were isolated and infected with recombinant HBV (rHBV) or wt HBV. GFP expression was monitored by confocal and flow cytometry. HBV DNA and HBV surface antigen (HBSAg) were analyzed by PCR and ELISA.

RESULTS: 3 × 107 wt HBV copies/mL and 5 × 106 rHBV copies/mL were collected from mice serum. In the wt HBV infected group, HBV progeny was 2 × 107 copies/mL and HBSAg was 770 ng/mL. In the rHBV infected group, GFP fluorescence was detected on d 3 post-infection and over 85% of the parenchymal cells expressed green fluorescence on d 12 post-infection. Compared with wt HBV in the PHH infection system, no rHBV DNA or HBSAg were detected in PHH culture media.

CONCLUSION: An effective HBV based vector was developed, which proved to be a useful HBV infection system. This vector and infection system can be applied to develop a therapeutic vector and study the HBV life cycle and viral pathogenesis.

Keywords: Hepatitis B virus, Primary human hepatocyte, Transfer plasmid, Helper plasmid, Infection system