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目的:初步探讨重组人白介素-27(interleukin-27,IL-27)体外对人外周血来源的细胞因子诱导的杀伤(cytokine induced killer,CIK)细胞增殖及其对肿瘤细胞的杀伤能力的影响。方法:无菌条件下分离健康志愿者外周血单个核细胞(peripheral blood mononuclear cell,PBMC),第0天加入IFN-γ、CD3单抗,之后根据加入不同细胞因子剂量随机分为六组进行CIK细胞培养:A组(IL-2:1 000 U/ml,正常对照组),B组(IL-2:1 000 U/ml,IL-27:20 ng/ml),C组(IL-2:1 000 U/ml,IL-27:10 ng/ml),D组(IL-2:500 U/ml,IL-27:10 ng/ml),E组(IL-2:1 000 U/ml,IL-27:5 ng/ml),F组(IL-2:1 000 U/ml,IL-27:40 ng/ml)。倒置相差显微镜下观察各组CIK细胞生长情况,流式细胞术检测各组CIK细胞CD3+CD56+T、CD8+T细胞的表达,自动细胞计数仪计数各组CIK细胞增殖情况,MTS方法测定各组CIK细胞对淋巴瘤K562细胞的杀伤活性。结果:培养第11天,D组与A、B、C组比较,CIK细胞中CD3+CD56+T细胞的表达[(66.57±2.44)%vs(60.03±1.75)%,(55.51±0.03)%,(56.07±0.83)%;均P<0.05]、CD8+T细胞的表达[(81.67±1.97)%vs(70.30±2.67)%,(74.92±2.47)%,(74.43±1.90)%;均P<0.05]都明显增强;D组CIK细胞培养的扩增倍数明显高于A、B、C组[(4 811.87±23.07)vs(3 257.73±91.97),(3790.92±64.49),(4 009.85±43.08)倍;均P<0.05];效靶比40∶1时;D组CIK细胞培养第11天时杀伤力为(76.71±2.21)%,显著高于其他各组(均P<0.05)。结论:细胞因子IL-27体外可显著提高CIK细胞的增殖能力和杀伤能力,最佳培养周期为11 d。
OBJECTIVE: To investigate the effect of recombinant human interleukin-27 (IL-27) on proliferation of human peripheral blood-derived cytokine induced killer (CIK) cells and its cytotoxicity on tumor cells in vitro. Methods: Peripheral blood mononuclear cells (PBMCs) were isolated from healthy volunteers under aseptic conditions. IFN-γ and CD3 monoclonal antibodies were added on day 0, then divided into six groups randomly according to different doses of cytokines for CIK Cell culture: Group A (IL-2: 1 000 U / ml, normal control group), group B (IL-2: 1000 U / ml, IL-27: 20 ng / (IL-2: 1000 U / ml, IL-27: 10 ng / ml) ml, IL-27: 5 ng / ml), group F (IL-2: 1000 U / ml, IL-27: 40 ng / ml). The growth of CIK cells in each group was observed under inverted phase contrast microscope. The expression of CD3 + CD56 + T and CD8 + T cells in CIK cells of each group was detected by flow cytometry. The proliferation of CIK cells in each group was measured by automatic cell counting system. Killing effect of CIK cells on lymphoma K562 cells. Results: On the eleventh day of culture, the expression of CD3 + CD56 + T cells in CIK cells in group D compared with those in groups A, B and C [(66.57 ± 2.44)% vs (60.03 ± 1.75)%, (55.51 ± 0.03) , And (56.07 ± 0.83)%, respectively; all P <0.05], and the expression of CD8 + T cells P <0.05]. The multiplication of CIK cell culture in group D was significantly higher than that in group A, B and C [(4 811.87 ± 23.07 vs 3 257.73 ± 91.97, 3790.92 ± 64.49, (4 009.85 ± 43.08) times; all P <0.05]. When the target ratio was 40: 1, the cytotoxicity of CIK cells in group D was (76.71 ± 2.21)% on day 11, significantly higher than that in other groups (all P <0.05). Conclusion: The cytokine IL-27 can significantly enhance the proliferation and cytotoxicity of CIK cells in vitro. The optimal culture period is 11 days.