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目的:探讨细胞因子IL-21对CIK细胞体外杀伤食管癌EC9706细胞活性的影响,并初步分析其可能的分子机制。方法:体外无菌分离人外周血单个核细胞,分别进行常规培养和加IL-21培养CIK细胞。流式细胞术检测2种培养CIK细胞的免疫表型,LDH释放法检测2种培养CIK细胞杀伤食管癌EC9706的活性,ELISA法检测2种CIK细胞培养上清液IFN-γ浓度。结果:常规培养和加IL-21培养的2种CIK细胞增殖数量无明显差异。加IL-21培养的CIK细胞CD3~+CD56~+细胞比例、CD3~+细胞内颗粒酶B和穿孔素表达率均显著高于常规培养CIK细胞[(26.95±3.53)%vs(16.18±1.04)%,(33.29±2.30)%vs(23.58±2.28)%和(59.70±1.91)%vs(45.96±2.67)%,均P<0.05)。效靶比20∶1和30∶1时,加IL-21培养的CIK细胞杀伤EC9706细胞的活性均显著高于常规培养的CIK细胞[20∶1时:(43.66±1.99)%vs(34.59±1.75)%;30∶1时:(57.52±2.15)%vs(42.23±1.87)%,均P<0.05]。加IL-21培养CIK细胞的上清液FN-γ的浓度明显高于常规培养CIK细胞的上清液[(142.7±13.4)vs(42.6±3.3)ng/L,P<0.05]。结论:IL-21增加CD3+CD56+细胞比例和颗粒酶B及穿孔素的表达,提高CIK细胞分泌IFN-γ,从而增强CIK细胞对EC9706细胞的杀伤活性。
Objective: To investigate the effect of cytokine IL-21 on the cytotoxicity of CIK cells in vitro against esophageal carcinoma EC9706 cells, and to analyze its possible molecular mechanism. Methods: Peripheral blood mononuclear cells were aseptically isolated from human peripheral blood and cultured in vitro. IL-21 and CIK cells were cultured in vitro. The immunophenotypes of two kinds of CIK cells were detected by flow cytometry. The activity of CIK cells against EC9706 was detected by LDH release assay. The concentrations of IFN-γ in two CIK cell culture supernatants were detected by ELISA. Results: There was no significant difference in the proliferation of two CIK cells cultured in routine culture and IL-21 culture. The percentage of CD3 ~ + CD56 ~ + cells and the expression of granzyme B and perforin in CD3 ~ + cells of CIK cells cultured with IL-21 were significantly higher than those of CIK cells [(26.95 ± 3.53)% vs (16.18 ± 1.04 ), (33.29 ± 2.30)% vs (23.58 ± 2.28)% and (59.70 ± 1.91)% vs (45.96 ± 2.67)%, all P <0.05). The cytotoxic activity of CIK cells cultured with IL-21 against EC9706 cells was significantly higher than that of the normal cultured CIK cells [20: 1 (43.66 ± 1.99)% vs (34.59 ± 1.75)%; at 30: 1: (57.52 ± 2.15)% vs (42.23 ± 1.87)%, all P <0.05]. The concentration of FN-γ in the supernatant of CIK cells cultured with IL-21 was significantly higher than that of the CIK cells ([142.7 ± 13.4] vs (42.6 ± 3.3) ng / L, P <0.05]. CONCLUSION: IL-21 increases the ratio of CD3 + CD56 + cells and the expression of granzyme B and perforin, increases the secretion of IFN-γ by CIK cells, and enhances the killing activity of CIK cells to EC9706 cells.