Studies Of Effect Of Adenouvirus-mediated HBx Transfaction On Lymphocyte In Vitro

Objective:To explore the amplification of HBx gene in normal human peripheral b-lood lymphocyte and the effect of adenouvirus-mediated HBx on the secretion of IL-10and lymphocyte apoptosis in vitro.Methods:As the treatment group, lymphocytes could be transfected with Ad-HBx which could be called the Ad-HBx/Lymphocyte group in vitro. Lymphocyte transfected with Ad-EGFP and non-transfected lymphocyte were cultured as the controls which were called the Ad-EGFP/Lymphocyte group and the Lymphocyte group in vitro. Q-PCR examination was used to detect the amplification of HBx gene in lymphocyte after culture for48h. Elisa was used to exmine the secretion of IL-10after culture for12h,24h,36h,48h. The lymphocyte apoptosis could be detected by Guava Nexin Reagent flow cytometry.Results:In vitro, there was a significant amplification of HBx gene in lymphocyte in the Ad-HBx/Lymphocyte group, and statistically significant differences compared with the Ad-EGFP/Lymphocyte group and the Lymphocyte group(P<0.05). Elisa demonstated that the concentration of IL-10in the Ad-HBx/Lymphocyte group was significantly lower than the Ad-EGFP/Lymphocyte group and the Lymphocyte group after culture for24h,36h,48h(P<0.05). Flow cytometry showed that the number of apoptotic cells in an early stage in experimental group were more than in the other two groups(P<0.05).Conclusion:There was a significant amplification of HBx gene in normal human pe-ripheral blood lymphocyte in vitro. Ad-HBx infection could effectively decrease the secretion of IL-10and promote lymphocyte apoptosis in an early stage. Effect of HBx on the regulation of the human immune system may be achieved by HBx gene expression in lymphocyte, the effect on the expression of anti-inflammatory factors and the promotion of early apoptosis.