IL-23单独或联合IL-2对人PBMC增殖及抗白血病活性影响的研究

The effects of IL-23 alone or combined with IL-2 on the proliferation and antileukemia activity of PBMC

目的比较单独应用IL-23或联合应用IL-23和IL-2对人外周血单个核细胞(PBMC)增殖及抗白血病活性的影响。方法密度梯度离心法分离人PBMC,以不同的细胞因子培养液培养,MTT比色法测定PBMC的增殖情况及PBMC对白血病K562细胞株的杀伤活性,流式细胞术分析诱导前后PBMC的细胞表型变化。结果细胞因子各组培养PBMC 1 d、3 d、5 d后,PBMC的增殖以及对K562细胞的杀伤活性均增强(P<0.05),IL-23与IL-2联合后PBMC的增殖以及杀伤活性进一步增强(P<0.05);IL-23(50 ng/ml)与IL-23+IL-2(50 ng/ml+100 IU/ml)组作用于PBMC 5 d后,检测CD3+细胞为(80.7±4.37)%和(83.2±4.04)%,CD16+CD56+细胞为(8.4±0.28)%和(12.7±0.9)%,CD4+细胞为(45.2±1.4)%和(47.0±1.72)%,CD8+细胞为(34.5±2.53)%和(35.4±2.11)%;与对照组相比,两组对PBMC细胞表型的增加有显著性差异(P<0.05);两组间比较CD16+CD56+细胞的表达上有显著性差异(P<0.05),对CD3+、CD4+、CD8+细胞的表达以及CD4+/CD8+比值的变化无显著性差异(P>0.05)。结论 IL-23对PBMC的增殖,以及PBMC对K562细胞的杀伤均有促进作用,与IL-2联合有协同作用。IL-23单独或联合IL-2诱导后PBMC中CD3、CD16/56、CD4、CD8抗原的表达均增加,二者联合后可以进一步增加CD16/56抗原的表达。

We aim to investigate the effects of IL-23 alone or combined with IL-2 on the proliferation and anti-leukemia activity of human peripheral blood mononuclear cells（PBMC）.PBMCs were separated by ficoll density gradient centrifugation,and the proliferation and cytotoxic activity against K562 were investigated by MTT assay.Changes in cell phenotype of PBMC were detected by Flow cytometry.PBMC’s proliferation and cytotoxic activity against K562 cells was enhanced in experiment groups after treatment for 1,3,5 days,which could be synergized by IL-23 plus IL-2 had.Surface markers of PBMC were significantly increased（P 〈 0.05） after treatment with IL-23（50 ng/ml） or IL-23（50 ng/ml） ＋IL-2（100 IU/ml） for 5 days： CD3＋ cells were（80.7±4.37）% and（83.2±4.04）%,CD16＋CD56＋ cells were（8.4±0.28）% and（12.7±0.9）%,CD4＋ cells were（45.2±1.4）% and（47.0±1.72）%,CD8＋ cells were（34.5±2.53）% and（35.4±2.11）%.The expression of CD16＋CD56＋ cells demonstrated significant difference between the two groups（P 〈 0.05）,but there was no significant difference of the expression of CD3＋ cells,CD4＋ cells,and CD8＋ cells between the two groups（P 〉 0.05）.The result suggested that IL-23 could enhance PBMC’s proliferation and cytotoxic activity,especially combined with IL-2.Furthermore,L-23 alone or combined with IL-2 could increase surface markers of PBMC,and IL-23 combined with IL-2 can further enhance the antigen expression of CD16/56.