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目的研究Toll样受体7(toll-like receptors 7,TLR7)激动剂(Tlr7a)对人细胞因子诱导的杀伤细胞(cytokine-inducedkiller cell,CIK)杀瘤效果的影响。方法采集3名健康志愿者外周血,分离外周血单个核细胞(peripheral blood mononuclear cell,PBMC),按照常规CIK细胞培养(Control组)或day0额外添加Tlr7a(Tlr7a组)2种方案培养2周。采用细胞计数法检测细胞的增殖效率,流式细胞术检测细胞亚群的比例,CCK-8比色法检测CIK细胞对人红白血病细胞(K562细胞)的杀伤活性。结果与对照比,Tlr7a处理后,总细胞中主要效应细胞CD3+~CD56+~双阳细胞占比增加4.1%,分别为(11.9±2.9)%和(16.0±5.8)%。杀伤实验结果显示,当E∶T为20∶1时,2组细胞对K562杀伤率分别为(98.8±4.2)%和(74.4±12.3)%。相比于Control组,Tlr7a组的细胞杀伤功能明显增强(P<0.05)。结论 TLR7激动剂(Tlr7a)具有增强CIK效应细胞的扩增能力,其刺激后获得的CIK细胞对K562肿瘤细胞的杀伤效果更好。
Objective To investigate the effect of toll-like receptor 7 (TLR7) agonist (Tlr7a) on the cytotoxicity of cytokine-induced killer cell (CIK). Methods Peripheral blood of 3 healthy volunteers were collected and peripheral blood mononuclear cells (PBMCs) were isolated and cultured for 2 weeks according to the two programs of conventional CIK cell culture (Control group) or day0 additionally added Tlr7a group (Tlr7a group). The cell proliferation efficiency was detected by cell counting method. The proportion of cell subsets was determined by flow cytometry. The killing activity of CIK cells on human erythroleukemia cells (K562 cells) was detected by CCK-8 colorimetric assay. Results Compared with the control, the proportion of CD3 + ~ CD56 + ~ double positive cells in total cells increased by 4.1% (11.9 ± 2.9)% and (16.0 ± 5.8)% respectively after Tlr7a treatment. The results of killing experiments showed that when the ratio of E: T was 20:1, the killing rates of K562 cells in two groups were (98.8 ± 4.2)% and (74.4 ± 12.3)%, respectively. The cytotoxicity of Tlr7a group was significantly higher than that of Control group (P <0.05). Conclusion The TLR7 agonist (Tlr7a) has the ability to enhance the expansion of CIK effector cells, and the cytotoxicity of CIK cells obtained after stimulation on K562 tumor cells is better.