本文選題：細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞 + 共刺激基因工程細(xì)胞��； 參考：《東南大學(xué)學(xué)報(醫(yī)學(xué)版)》2017年05期

【摘要】：目的:研究共刺激基因工程細(xì)胞共培養(yǎng)的細(xì)胞因子誘導(dǎo)的殺傷細(xì)胞(ECCE-CIK)對人肝癌細(xì)胞系Hep G2.2.15細(xì)胞中乙肝病毒的抑制作用。方法:將共刺激基因工程細(xì)胞與人外周血單個核細(xì)胞共同培養(yǎng),使用IFN-γ、CD3單抗、IL-2細(xì)胞因子誘導(dǎo),產(chǎn)生ECCE-CIK細(xì)胞;將效應(yīng)細(xì)胞(ECCE-CIK)和靶細(xì)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染色法檢測ECCE-CIK對Hep G2.2.15的殺傷作用;建立體外直接法與間接法共培養(yǎng)系統(tǒng),收集作用后3、24、48 h的培養(yǎng)上清液,檢測HBV DNA與HBsAg水平。結(jié)果:隨著效靶比的增加和時間的延長,殺傷率逐漸上升,培養(yǎng)上清中HBV DNA、HBsAg水平逐漸下降(P0.05);直接系統(tǒng)對HBV DNA、HBsAg的抑制曲線略高于間接系統(tǒng)。結(jié)論:在體外直接法與間接法共培養(yǎng)系統(tǒng)中,ECCE-CIK都能降低Hep G2.2.15中HBV DNA和HBsAg水平,提示ECCE-CIK能通過細(xì)胞毒和非細(xì)胞毒方式抑制靶細(xì)胞中乙肝病毒的復(fù)制,為臨床治療提供了實驗依據(jù)。
[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- 緯 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.
【作者單位】： 東南大學(xué)附屬第二醫(yī)院;丹陽市人民醫(yī)院;
【基金】：國家自然科學(xué)基金資助項目(81402559)
【分類號】：R373.21

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