IL-2、IL-21诱导人外周血单个核细胞及其抗肿瘤作用

摘要

Objective To investigate the changes of proliferation, phenotype and in vitro antitumor activity of human peripheral blood mononuclear cells (PBMC) induced by IL-2 and IL-21. Methods Sort PBMCs into 4 gropus in the concentration of 1 × 106/ml, IL-2 of 100 IU/ml was added into IL-2 group, IL-21 of 100 IU/ml was added into IL-21 group,IL-2 of 50 IU/ml and IL-21 of 50 IU/ml was added into joint group. Saline solution of 0. 1 ml was added into the control group. Flow cytometry assay was used to analysis the subsets of PBMC after being induced and Trypan blue staining of living cells was counted in each group. The above groups was taken as effector cells (E), the logarithmic phase of 4 kinds of tumor cells ( human gastric cancer cell line M85, gastric cancer cell lines BGC823, colon cancer cell line HCT116, colorectal adenocarcinoma cell line HCT8) as target cells (T), dilute targets cells into 5 × 103 cells/well,the ratio of E: T was 1:1 and2:l. MTT assay was used to detect the cytotoxic effects( killing ratio). Results Live cell counts in the joint group was higher than that tin the IL-2 group,IL-21 group and control group,all P ＜0.05;CD3-/CD56+ ,CD3+/CD56+ in the joint group, IL-2 group and IL-21 group were higher than those in the control group, P ＜ 0. 05 or 0.01. The killing ratio of 4 kinds of tumor cells in the joint group was higher than the other three groups, and which in IL-2 group, IL-21 group was higher than that in the control group, P ＜ 0. 05. Conclusion IL-2 combined synergistic stimulation of IL-21 can effectively stimulate PBMC proliferation and phenotypic changes ,and can kill 4 kinds of digestive tract cancer cell lines strongly.%目的 探讨IL-2、IL-21对人外周血单个核细胞(PBMC)的诱导增殖和表型改变的影响及其体外抗瘤作用.方法 取PBMC细胞,调整浓度为1×106个/ml,分为四组,IL-2组加入IL-2 100 IU/ml,IL-21组加入IL-21 100 IU/ml,联合组加入IL-2 50 IU/ml和IL-21 50 IU/ml,对照组加入0.1 ml生理盐水,台盼蓝染色法计数各组活细胞,流式细胞仪检测各组PBMC表型;以上述四组为效应细胞(E),以对数生长期的4种肿瘤细胞(人胃癌细胞系M85,胃癌细胞系BGC823,结肠癌细胞系HCT116,结直肠腺癌细胞系HCT8)为靶细胞(T),稀释靶细胞5×103个/孔,E:T为1:1和2:1,MTT法测定细胞毒性(杀伤率).结果 联合组活细胞计数高于IL-2组、IL-21组、对照组,P均＜0.05;联合组、IL-2组和IL-21组CD3 -/CD56+、CD3 +/CD56 +水平高于对照组,P＜0.05或0.01;联合组对4种肿瘤细胞的杀伤率均高于其余三组,IL-2组、IL-21组高于对照组,P均＜0.05.结论 IL-2联合IL-21的协同刺激作用,可有效地刺激PBMC的增殖和表型改变,对4种消化道肿瘤细胞株均有较强的杀伤力.