共刺激基因工程细胞共培养的细胞因子诱导的杀伤细胞对HepG2.2.15细胞中乙肝病毒的抑制作用研究

发布时间：2018-06-05 08:50

本文选题：细胞因子诱导的杀伤细胞 + 共刺激基因工程细胞　；参考：《东南大学学报(医学版)》2017年05期

【摘要】：目的:研究共刺激基因工程细胞共培养的细胞因子诱导的杀伤细胞(ECCE-CIK)对人肝癌细胞系Hep G2.2.15细胞中乙肝病毒的抑制作用。方法:将共刺激基因工程细胞与人外周血单个核细胞共同培养,使用IFN-γ、CD3单抗、IL-2细胞因子诱导,产生ECCE-CIK细胞;将效应细胞(ECCE-CIK)和靶细胞(Hep G2.2.15)以效∶靶1∶1、5∶1、20∶1的比例共同孵育,使用CFSE/PI染色法检测ECCE-CIK对Hep G2.2.15的杀伤作用;建立体外直接法与间接法共培养系统,收集作用后3、24、48 h的培养上清液,检测HBV DNA与HBsAg水平。结果:随着效靶比的增加和时间的延长,杀伤率逐渐上升,培养上清中HBV DNA、HBsAg水平逐渐下降(P0.05);直接系统对HBV DNA、HBsAg的抑制曲线略高于间接系统。结论:在体外直接法与间接法共培养系统中,ECCE-CIK都能降低Hep G2.2.15中HBV DNA和HBsAg水平,提示ECCE-CIK能通过细胞毒和非细胞毒方式抑制靶细胞中乙肝病毒的复制,为临床治疗提供了实验依据。
[Abstract]:Aim: to study the inhibitory effect of costimulatory gene engineering cells co-cultured by cytokine induced killer cell line ECCE-CIK on hepatitis B virus (HBV) in human hepatoma cell line Hep G2.2.15. Methods: the co-stimulated engineering cells and human peripheral blood mononuclear cells were co-cultured and induced by IFN- 纬 CD3 monoclonal antibody IL-2 cytokines to produce ECCE-CIK cells. ECCE-CIK) and Hep G2.2.15) were incubated with the ratio of target 1: 1: 5: 1 to 20: 1. The killing effect of ECCE-CIK on Hep G2.2.15 was detected by CFSE/PI staining, and the system of direct and indirect co-culture was established. The supernatant of culture was collected for 48 h and the levels of HBV DNA and HBsAg were detected. Results: with the increase of the effect-target ratio and the prolongation of the time, the killing rate gradually increased, and the level of HBV in the supernatant decreased gradually, and the inhibition curve of the direct system on HBV was slightly higher than that on the indirect system. Conclusion: ECCE-CIK can decrease the levels of HBV DNA and HBsAg in Hep G2.2.15 in both direct and indirect co-culture system in vitro, suggesting that ECCE-CIK can inhibit HBV replication in target cells by cytotoxicity and non-cytotoxicity, and provide experimental evidence for clinical treatment.
【作者单位】：东南大学附属第二医院;丹阳市人民医院;
【基金】：国家自然科学基金资助项目(81402559)
【分类号】：R373.21

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