Characterization of hepatitis B virus antigen expression in vitro and in vivo transduced by different transfer plasmids carrying HBV infectious genome / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>To character HBV antigen expression in vitro and in vivo transduced by different transgenic plasmids carrying infectious genome of hepatitis B virus (HBV).</p><p><b>METHODS</b>We 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.</p><p><b>RESULTS</b>We 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).</p><p><b>CONCLUSION</b>The 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.</p>