目的：探讨当归多糖（ Angelica sinensis polysaccharide，ASP）体外对人外周血来源的细胞因子诱导的杀伤（cytokine induced killer，CIK）细胞的增殖、免疫表型及其对白血病K562细胞杀伤活性的影响。方法：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 (1 000 IU/ml)，24 h后再加入IL-2 (1 000 IU/ml)、抗CD3单抗 (50 ng/ml) 继续培养，此后每隔3 d根据添加不同浓度的ASP分为5组：A组（不加ASP）、B～E组（分别加ASP 12.5、25、50、100 μg/ml）。全自动血细胞计数仪计数各组CIK细胞的增值情况，流式细胞术检测各组CIK细胞中CD3+CD4+、CD3+CD8+、CD3+CD56+细胞的百分比，酶联免疫吸附（ELISA）法检测各组CIK细胞上清液中IFN-γ及TNF-α的分泌水平，CCK-8法检测各组CIK细胞对K562细胞的杀伤活性。 结果：联合ASP诱导培养CIK细胞至第16天时，与A组相比，E组CIK细胞的增殖能力明显增加\[（56.98 ± 3.00）×106 vs （43.81 ± 2.14）× 106， P <0.05）\]，各组细胞活率都保持在90%以上；各组CIK细胞CD3+CD4+、CD3+CD8+细胞比例无明显差异，而D组、E组CD3+CD56+细胞比例与A组相比存在差异\[（26.65±3.71）%、（28.36±4.28）% vs （20.75±3.56）%， P <0.05）\]；E组CIK细胞在效靶比为40 ∶1、20 ∶1、10 ∶1时对K562细胞的杀伤活性比A组更强\[（84.19±5.88）% vs （68.05±5.95）%、（76.69±6.54）% vs （64.55±9.44）%、（72.32±9.22）% vs （61.45±8.22）%，均 P <0.05]。结论：随着培养时间的延长，高浓度的ASP体外对CIK细胞的增殖及杀伤活性有一定的促进作用。

Objective: To characterize the biological features of Angelica sinensis polysaccharide (ASP) and evaluate its effect on the proliferation, immunotype and cytotoxicity of cytokine induced killer cells (CIK) derived from human peripheral blood mononuclear cells (PBMCs). Methods: PBMCs were obtained from healthy adult donors and induced to differentiate into CIK cells with interferon-γ (1 000 IU/ml) and anti-human CD3 (50 ng/ml). CIK cells were then treated with ASP every 3 days at 0, 12.5, 25, 50 and 100 μg/ml, respectively. At 10 days after treatment, the number of CIK cells was counted by computer-based cell counter, the proportion of CD3+CD4+, CD3+CD8+ and CD3+CD56+ cells were analyzed by flow cytometry , and the cytotoxicity of CIK cells were tested by CCK-8 method. Results: At the end of 16 day treatment, above 90% of CIK cells remained viable in all treatment groups, and the proliferative activity was significantly higher ( P <0.05) in CIK cells treated with 100 μg/ml ASP (56.98±3.00)×106 than in cells treated with 0 μg/ml ASP (43.81±2.14)×106. While the CD3+CD56+ cell ratio was significantly increased ( P <0.05) in CIK cells treated with 50 μg/ml ASP (26.65±3.71)% or 100 μg/ml ASP(28.36±4.28%) as compared with the control（20.75±3.56)% , there were no statistical differences in the proportion of CD3+CD4+ or CD3+CD8+ cells among the different groups ( P >0.05). The cytotoxic activity of CIK cells against K562 cells was significantly increased ( P <0.05) after treatment with 100 μg/ml ASP as compared with 0 μg/ml ASP at an effector-target ratio of 40∶1 (84.19±5.88)% vs （8.05±5.95)%, 20∶1（76.69±6.54)% vs (64.55±9.44)% or 10:1（72.32±9.22)% vs (61.45±8.22)%. Conclusion: Angelica sinensis polysaccharide can enhance the proliferative and cytotoxic activities of CIK cells in dose- and time-dependent manners.

甘肃省科技重大专项（No. 1102FKDA005）