[Construction of a eukaryotic expression vector expressing human IFN-gamma and its inhibitory effect on HBV replication in vitro].
OBJECTIVE
To observe the inhibitory effect of a eukaryotic expression vector expressing human IFN-gamma (pcDNA3.1- IFN-gamma) on HBV replication in hepG2.2.15 cells.


METHODS
The eukaryotic expression vector expressing human IFN-gamma was constructed using PCR and gene recombination technique. hepG2.2.15 cells were transfected with pcDNA3.1-IFN-gamma and the culture supernatant was collected to determine the expression of IFN-gamma protein by ELISA. The HBV DNA copies and the concentration of HBeAg and HBsAg were measured by fluorescence real-time PCR and ELISA kit, respectively.


RESULTS
Compared with that of negative control and blank 2.2.15 cells, the concentration of HBeAg in the supernatant of 2.2.15 cells transfected with pcDNA3.1- IFN-gamma were decreased by 49%, and HBsAg concentration was lowered by 35% and 33%, respectively. A significant decrease of HBV DNA copies was observed in pcDNA3.1- IFN-gamma-transfected cells in comparison with the two control cells. No significant differences were noted in all the results between the two control groups.


CONCLUSIONS
We have successfully constructed the eukaryotic expression vector expressing human IFN-gamma, which provides a basis for anti-HBV gene therapy using human IFN-gamma.
Li
Duo-lu
DL
First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China. dorali1979@yahoo.com.cn
Kan
Quan-cheng
QC
D005822:Genetic Vectors; D056945:Hep G2 Cells; D006515:Hepatitis B virus / Q000187:drug effects*; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics* / Q000494:pharmacology*; D014162:Transfection; D014779:Virus Replication / Q000187:drug effects
D007371:Interferon-gamma