乙型肝炎病毒全基因重组腺病毒感染免疫研究

目的 用腺病毒载体将乙型肝炎病毒(HBV)基因组导入小鼠体内,研究HBV表达产物诱发的机体免疫反应.方法 用AdEasy系统构建含HBV基因组的重组腺病毒AdHBV-WT,感染HepG2细胞后,分别用ELISA和western blot检测细胞培养上清液和细胞裂解物中的HBsAg、HBeAg和HBcAg.AdHBV-WT经在293细胞中扩增、纯化和浓缩后,经滴鼻途径感染BALB/C小鼠,检测体液和细胞免疫反应.结果 重组腺病毒AdHBV-WT感染HepG2细胞后,有效地表达出HBV的各种抗原.重组腺病毒感染小鼠可诱导抗-HBc的产生.3H-TdR摄入实验表明,AdHBV-WT可诱发小鼠产生HBcAg特异性的T细胞应答.结论 HBV全基因重组腺病毒在体外可表达HBV的各种抗原,并可诱发小鼠产生HBcAg特异性的体液免疫和细胞免疫.

Study on the immune response evoked by recombinant adenovirus containing HBV genome

Objective HBV genome was transferred into mice to study the immune response evoked by HBV expression production. Methods AdHBV-WT, the recombinant adenovirus containing HBV genome, was constructed using AdEasy system, then infected HepG2 cell lines. The HBV antigens expression in cell culture in vitro was demonstrated by ELISA and Western blot. After amplification, purification and concentration, BALB/C mice were infected by AdHBV-WT intranasally. The immune responses were measured by ELISA for antibody responses and ^3 H-TdR uptake assay for T cell prolification. Results The recombinant adenovirus AdHBV-WT expressed HBV antigens, HBsAg, HBeAg and HBcAg effectively in HepG2 cell lines. Anti-HBc antibody was detected in mice sera in experimental group. Furthermore, T cell proliferation response to HBcAg was also induced by AdHBV-WT, which was confLrmed by ^3 H-TdR uptake assay. Conclusion AdHBV-WT could express HBV antigens effectively in cell cuhure and induced humoral and cellular immunization in BALB/C mice.