Viral Hepatitis
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. May 7, 2005; 11(17): 2574-2578
Published online May 7, 2005. doi: 10.3748/wjg.v11.i17.2574
Anti-HBV activity of TRL mediated by recombinant adenovirus
Wei-Dong Gong, Ya Zhao, Jun Yi, Jin Ding, Jun Liu, Cai-Fang Xue
Wei-Dong Gong, Department of Oncology, Zhujiang Hospital, Nanfang Medical University, Guangzhou 510280, Guangdong Province, China
Ya Zhao, Jin Ding, Jun Liu, Cai-Fang Xue, Department Pathogenic Organisms, Fourth Military Medical University, 710033, Shaanxi Province, China
Jun Yi, Department of Vascular Surgery, Xijing Hospital, Military Medical University, Xi抋n 710033, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Supported by the National Natural Scientific Foundation, No. 30400380 and Shaanxi Natural Scientific Foundation, No. 22C2-28
Correspondence to: Wei-Dong Gong, Department of Oncology, Zhujiang Hospital, Nanfang Medical University, Guangzhou 510280, Guangdong Province, China. gongwd70@fmmu.edu
Telephone: +86-29-83373773 Fax: +86-29-83374199
Received: November 8, 2004
Revised: November 9, 2004
Accepted: December 21, 2004
Published online: May 7, 2005
Abstract

AIM: To investigate the inhibitive effect of hepatitis B virus (HBV)-TRL on HBV replication.

METHODS: Based on previously constructed pcDNA3.1(-)/TRL, TR, TRmut, HBV core protein (HBVc) and hEDN, interest gene sequences TRL, TR, HBVc and hEDN were inserted into adenovirus shuttle plasmid pDC316 respectively and co-transfected HEK293 cells with rescue plasmid pBHGlox(delta)E1,3Cre to acquire RAd/TRL, TR, HBVc and hEDN. And then RAds were identified, amplified and the titers in HEK293 cells were determined. RAd/TRL and TR were named as the experimental groups, and others were control ones. After HepG2.2.15 cells were infected, RAd/TRL expression was identified by indirect immunofluorescence staining. Supernatant HBV-DNA content was determined by fluorescent quantification PCR. Meanwhile, metabolism of HepG2.2.15 cells was evaluated by MTT colorimetry.

RESULTS: RAd vectors with distinct interest gene sequence were successfully constructed. Effective expression of RAd/TRL in HepG2.2.15 cells resulted in a significant decrease of supernatant HBV-DNA content compared to RAd/TR (0.63±0.14 vs 1.60±0.47, P = 0.0266, <0.05) and other control groups (0.63±0.14 vs 8.50±2.78, 8.25±2.26, 8.25±2.29, 8.50±1.51, 8.57±1.63, P<0.01). MTT assay suggested that there were no significant differences in cell metabolic activity between groups (P>0.05).

CONCLUSION: The construction and expression of RAd/TRL has been achieved and it could inhibit HBV replication successfully, which has laid the foundation for further research on anti-HBV activity in vivo.

Keywords: HBV, HBV-TRL