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目的研究不同细胞因子诱导的树突状细胞(DC)体外抗肿瘤效应。方法常规分离健康人外周血单核细胞,分别采用粒巨噬集落刺激因子(GM-CSF),白细胞介素(IL)-4,肿瘤坏死因子(TNF),GM-CSF、IL-4、α干扰素(IFN-α)细胞因子组合将其诱导为DC,并分别负载SMMC7721细胞冻融抗原;用流式细胞仪检测第9天时细胞表面HLA-DR、CD1α、CD80、CD83的表达;采用四甲基偶氮唑盐比色(MTT)法检测各组刺激自体淋巴细胞增殖能力,并检测各组DC诱导的细胞毒性T淋巴细胞(CTL)对SMMC7721细胞株的杀伤率。结果第9天显微镜下观察各组细胞,其细胞形态无明显差异,但流式细胞仪显示IFN-α组DC其膜表面标志CD80、CD83、CD1a表达上调,在刺激同种T细胞增殖能力方面,较TNF-α组DC效果明显。且IL-12分泌明显增加。在肝癌细胞株SMMC7221杀伤实验中,培养液中加入IFN-α后杀伤效应明显。结论与TNF-α相比,IFN-α诱导DC一定程度上能增强其特异性杀伤能力。
Objective To study the antitumor effects of dendritic cells (DCs) induced by different cytokines in vitro. Methods Normal human peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers. GM-CSF, IL-4, TNF, GM-CSF and IL- Interferon-α (IFN-α) cytokines were used to induce DCs and were respectively loaded with freeze-thaw antigens of SMMC7721 cells. Flow cytometry was used to detect the expression of HLA-DR, CD1α, CD80 and CD83 on day 9, Methyl thiazolyl tetrazolium (MTT) assay was used to detect the ability of each group to stimulate autologous lymphocyte proliferation. The cytotoxic T lymphocyte (CTL) -induced cytotoxicity of each group on SMMC7721 cell line was tested. Results On the 9th day, the morphological changes of the cells in each group were observed under the microscope. However, the expression of CD80, CD83 and CD1a on the membrane surface of DCs in IFN-α group was up-regulated by flow cytometry. , Compared with TNF-α group DC effect is obvious. And IL-12 secretion increased significantly. In the anti-SMMC7221 hepatocellular carcinoma cell line, the killing effect was obvious when IFN-α was added to the culture medium. Conclusion Compared with TNF-α, IFN-α-induced DC can enhance its specific killing ability to a certain extent.