自体特异性免疫效应淋巴细胞抑制转HBV基因小鼠HBV复制的研究

Study on effect of autologous specific immune effective cells on inhibiting HBV replication in HBV transgenic mice

目的:探讨经乙型肝炎病毒(HBV)转基因小鼠树突状细胞(DCs)体外诱导的特异性免疫效应细胞(IECs)对自体HBV复制的影响。方法:从转HBV基因小鼠体内提取DCs,体外诱导为成熟DCs,与淋巴细胞共培养,诱导为特异性IECs,将其经阴茎背静脉注入小鼠体内。实验分为2组:生理盐水(NS)组、IEC组,观察6个时间点:0 h、2、4、6、8周和12周;通过生化检测肝功能,PCR检测血清中HBV DNA水平、ELISA检测细胞因子、免疫组化检测肝组织内的HBs Ag和HBc Ag,评估IECs对小鼠体内HBV复制的影响。结果:IEC组小鼠于6、8周和12周时,肝功能明显改善,HBV DNA水平明显降低,HBs Ag和HBc Ag明显减少,且均优于NS组(P<0.05)。结论:特异性IECs可修复转HBV基因小鼠肝脏功能、抑制HBV复制,相关的细胞因子参与其作用。

Objective： To investigate the effect of specific immune effective cells （IECs） induced by dendritic cells （DCs） from HBV transgenic mice on HBV replication. Methods： DCs were isolated from HBV transgenic mice, induced into mature DCs and co-cultured with lymphocytes to produce IECs, which were injected into transgenic mice by tentum dorsal veins. Samples were divided into two groups： normal saline （NS） group and IEC group, which were observed at 0 h, 2, 4, 6, 8 week and 12 week. To evaluate the effect of IECs, liver function were checked by biochemistry, the expression of HBV DNA were checked by PCR, the levels of cytokines were checked by ELISA, and HBsAg and HBeAg expression were checked by immunohistochemistry. Results： At 6, 8 week and 12 week after injection of IECs into transgenic mice, liver function were improved significantly. HBV DNA, HBsAg and HBcAg expression decreased significantly. And all indexes were better than NS group （P〈0.05）. Conclusion： IECs could protect liver function, and inhibit HBV replication in HBV transgenic mice, and the related cytokines are involved in this effect.