HBIG抑制HBsAg、HBV DNA分泌的体外实验研究
摘要
Objective To investigate in vitro transmembrane transportation of hepatitis Bimmunoglobulin (HBIG) through hepatocytes and inhibition of HBsAg release and HBV replication in hepatocyte-derived cell line. Methods QSG7701 cells were cultured with DMEM containing different concentrations of HBIG. At different time points after culture, the supernatant was collected for HBsAb quantitative detection by time-resolved immunofluorometric assay (IFMA) and the amount of intracellular HBIG was calculated. HepG 2.2.15 cells were cultured with DMEM in the absence or present of different concentrations of HBIG. The supernatant at some days interval was collected for HBsAg quantitative detection by IFMA and HBV DNA quantitative detection by reel-time fluorescence quantitative PCR (RTFQ-PCR). MTT assay was used to evaluate the cytotoxicity of HBIG. Result (1) 38%~46% of 0.1~0.4IU/mL HBIG was endocytosed into QSG7701 after 48 hours of culture. (2) HBIG in the concentration from 0.01IU/mL to 10.0IU/mL. had no cytotoxicity to the cells of QSG7701 and HepG2.2.15. (3) HBsAg and HBV DNA in supernatant of HepG2.2.15 cultured with 0.1~10.0IU/L HBIG reduced significantly than that of HepG2.2.15 cultured without HBIG after 3, 6, 9 days incubation, respectively (P＜0.01). (4) The amount of HBsAg secreted in supernatant reduced continuously at the 5th and 7th day after HBIG was removed, (P＜0.01), but rebounded at the 9th (P＜0.01) and 11th day after HBIG was removed, respectively, as compared with the control. The amount of HBV DNA secreted in supernatant reduced continuously at the 5th day after HBIG was re moved (P＜0.01), but rebounded at the 7th, 9th and 11th day after HBIG was removed, respectively, as compared with the control. Conclusion HBIG can be endocytosed into hepatocytes and extracellular or intracellular HBTG can inhibit the secretion of HBsAg and HBV DNA in vitro.