Antiª²virus activity of recombinant adenoviral vector HBVª²TRL

    GONG Weiª²Dong, YI Jun, ZHAO Ya, LIU Jun, DING Jin, XUE Caiª²Fang

    1Department of Scientific Research Administration, 2Department of Vascular Surgery, Xijing Hospital, 3Department of Pathogenic Organisms, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To investigate the inhibitive effect of HBVª²TRL on the HBV replication. METHODS: Based on previously constructed pcDNA3.1(-)/TRL, TR, TRmut, HBVc and hEDN, we inserted interest gene sequences TRL, TR, HBVc and hEDN respectively into adenovirus shuttle plasmid pDC316 and cotransfected HEK293 cells with rescue plasmid pBHGlox (delta)E1,3Cre to acquire RAd/TRL, TR, HBVc and hEDN. RAds were then identified, amplified and the titers were determined. RAd/TRL and TR were taken as the experimental groups and others were as control. After infecting HepG2.2.15 cells, RAd/TRL expression was identified by indirect immunofluorescence staining. The supernatant HBVª²DNA content was determined by fluorescent quantification PCR (FQª²PCR). The 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 HBVª²DNA content compared with that of RAd/TR (P=0.0266, P<0.05) and other control groups (P<0.01). MTT assay suggested that there were no significant differences between the groups (P>0.05). CONCLUSION: RAd/TRL is successfully constructed and its effective expression inhibits the HBV replication. ......