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。

bjective : To investigate the effect of IL-27 on the proliferation and cytotoxicity of cytokine induced killer (CIK) cells derived from human peripheral blood mononuclear cells (PBMC). Methods: Monocytes were purified from PBMCs obtained from healthy adults. After being challenged with interferon-γ (IFN-γ) and anti-human CD3, the cells were radomly divided into six groups by different stimulating factors: A group (IL-2：1 000 U/ml), B group (IL-2：1 000 U/ml, IL-27：20 ng/ml), C group (IL-2：1 000 U/ml, IL-27：10 ng/ml), D group (IL-2：500 U/ml, IL-27：10 ng/ml), E group (IL-2：1 000 U/ml, IL-27：5 ng/ml), and F group (IL-2：1000 U/ml, IL-27：40 ng/ml). The morphology of the CIK cells in different groups were evaluated by inverted microscopy. The proportion of CIK cells respectively expressing CD3 +CD56 + and CD8 + were analyzed by fluorescence activating cell sorter (FACS) on days 7 , 9, 11 and 13 after treatments. The number of attached cells was counted by a computer-based cell counter. The cytotoxicity of CIK cells was determined by MTS test. Results: Compared with A, B and C groups, D group had significantly higher proportions of CD3 +CD56 + CIK cells (\[66.57±2.44\]% vs \[60.03±1.75\]%, \[5551±0.03\]%, and \[56.07±083\]%; P<0.05) and CD8 + CIK cells (\[81.67±1.97\]% vs \[70.30±2.67\]%, \[74.92±2.47\]%, and \[74.43±1.90\]%; P<0.05), and significantly higher index of cell expansion to culture median (4 811.87±23.07 vs 3 25773±91.97, 3 790.92±64.49, 4 009.85±43.08; P<0.05) on day 11 after treatments. The highest effector to target cell ratio on day 11 in D group was 40 ∶1 with a cytotoxicity rate of (76.71±221)% which was significantly higher than that in groups A, B, and C. Conclusion: In vitro, IL-27 is capable of significantly enhancing the proliferation and cytotoxicity of CIK cells in a time-dependent manner and the optimal time of incubation seems to be 11 days.

国家自然科学基金资助项目（No.81201607），教育部博士点新教师基金资助项目（No.20101323120004），河北省自然科学基金资助项目（No.H2012206135）