IL-23单独或联合IL-2诱导人外周血单个核细胞对K562细胞杀伤效应的实验研究

IL-23 alone or with IL-2 Induces the Killing Effect of hPBMNC on K562 Cells

本研究旨在探讨IL-23单独或联合IL-2诱导的人外周血单个核细胞(PBMNC)对白血病细胞的杀伤效应及作用机制。IL-23(50 ng/ml)单独或联合IL-2(100 U/ml)体外诱导正常人PBMNC 72 h,并与白血病细胞株K562共同培养。采用CCK-8法测定不同时间诱导后的PBMNC对K562细胞的杀伤效应;采用ELISA方法检测杀伤活性最大时细胞培养液中IFN-γ的水平;应用RQ-PCR法检测诱导后PBMNC的穿孔素、颗粒酶B的表达水平。结果表明,IL-23单独或联合IL-2作用后的PBMNC均对K562细胞有杀伤活性,随着时间延长,杀伤率明显增加,各个时间点比较,有显著性差异(P<0.05);各细胞因子组培养液中分泌IFN-γ水平均显著高于空白对照组(P<0.05),其中以IL-23联合IL-2组诱导PBMNC表达IFN-γ的水平最高,与其它两组比较,差异显著(P<0.05)。各细胞因子组PBMNC的穿孔素、颗粒酶B mRNA的表达量均较对照组明显增加(P<0.05),且IL-23联合IL-2组的穿孔素、颗粒酶B mRNA表达量显著高于其他组(P<0.05)。结论:IL-23能促进PBMNC对K562细胞的杀伤活性,与IL-2联合具有协同增强作用,并呈时间依赖性。IL-23作用于PBMNC后IFN-γ、穿孔素、颗粒酶B的表达均明显增加,且与IL-2具有协同作用。推测IL-23可能通过诱导PBMNC表达IFN-γ、穿孔素、颗粒酶B发挥抗白血病细胞作用。

This study was aimed to explore the killing effect of PBMNC induced by IL-23 alone or combined with 1L-2 on K562 ceils and its mechanism. The PBMNC were induced in vitro by 1L-23 （50 ng/ml） alone or IL-23 combined with IL-2 （100 U/ml） for 72 h, and then were co-cultured with leukemia cell line K562. The CCK-8 method was used to detect the effect of PBMNC induced at different times on K562 cells, the ELISA was performed for detecting 1FN-γ level in culture supernatant, and the perforin and granzyes B were detected by RQ-PCR. The results showed that the killing effect of PBMNC induced by IL-23 alone or IL-23 combined with IL-2 on K562 cells was observed, and obviously enhaned with prolonging of time, moreover, there was statistical difference among different time points （P 〈 0.05 ）. The 1FN-γ level in supematant of PBMNC cultured with cytokines significantly increased, and the IFN-γ levels in group of IL-23 combined with IL-2 were higher than that in other groups （P 〈 0. 05）. The mRNA expressions level of perforin and granzymes B of the expanded PBMNC in groups cultured with cytokines were higher than that in control group （ P 〈 0.05 ）, and the mRNA expressions of perforin and granzymes B in group of IL-23 combined with IL-2 were significantly higher than that in others （ P 〈 0. 05 ）. It is concluded that IL-23 can promote the killing effect of PBMNC on K562 cells. The combination of IL-2 with IL-23 displays synergic effect and a time-dependent manner. IL-23 also enhances the expression of IFN-γ, perforin and granzyme B in PBMNC. Its combinetion with IL-2 displays synergistic effect, suggesting that the anti-leukemic activity of IL-23 may be realized through inducing PBMNC to express IFN-γ, perforin and granzyme B.