Characterization of hepatitis B virus antigen expression in vitro and in vivo transduced by different transfer plasmids carrying HBV infectious genome.
- Author:
Xia CHUAI
1
;
Hong CHEN
;
Yang YANG
;
Wen WANG
;
Bo WEN
;
Gang WANG
;
Bao-Ying HUANG
;
Yao DENG
;
Wen-Jie TAN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Disease Models, Animal; Female; Genome, Viral; Hepatitis B Antigens; genetics; Lentivirus; genetics; Mice; Mice, Inbred C57BL; Plasmids; Transduction, Genetic; methods
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(2):96-98
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo character HBV antigen expression in vitro and in vivo transduced by different transgenic plasmids carrying infectious genome of hepatitis B virus (HBV).
METHODSWe constructed four different lentiviral transfer plasmids (carrying 1.3 full-length genome of HBV, by replacing the EGFP express box in pCS-CG plasmid with HBV genome and with different structural element, named as pCS-HBV1.3 (pCS-HBV1.3 X, pCS-HBV1.3 P, pCS-HBV1.3 N and pCS-HBV1.3 K). We detected the expression of HBsAg and HBeAg by ELISA in different time after transfected Huh 7 cells or hydrodynamic injection into C57 BL/6 mice with transfer plasmids pCS-HBV, respectively.
RESULTSWe detected significant expression of HBsAg over 5 days after transfected Huh 7 cells (in vitro) or hydrodynamic injection into C57 BL/6 mice (in vivo) with transfer plasmids pCS-HBV1.3 X, pCS-HBV1.3 P and pCS-HBV1.3 K. The expression level and dynamics of HBsAg and HBeAg in the sera of mice is consistent with that of in the supernatant of Huh-7 cell. Furthermore, the expression of HBV antigens were modulated by the direction and position of HBV insert, also by some lentiviral vector cis-elements (cPPT and RRE).
CONCLUSIONThe optimal lentiviral transfer plasmids (pCS-HBV1.3 X, pCS-HBV1.3 P and pCS-HBV1.3 K) could be further used for establishment and application of HBV transgenic cell or animal model.
