摘要
目的研究调节性T细胞对外周血来源的DC-CIK细胞增殖及杀伤活性的影响。方法无菌分离外周血单个核细胞,体外诱导分化为DC-CIK细胞。实验分2组:DC-CIK组为常规单个核细胞诱导分化的DC-CIK;DCCIK-Tregdel组为培养前利用免疫磁珠分选去除CD4+CD25+T细胞的外周血单个核细胞诱导分化的DC-CIK。细胞扩增倍数法检测2组细胞的增殖能力;MTT法检测肿瘤细胞的杀伤活性;流式细胞仪检测免疫表型;ELISA法检测细胞因子分泌水平。结果培养14 d的DC-CIK-Tregdel组细胞的扩增倍数高于DC-CIK组,并且CD3+CD8+、CD3+CD56+双阳性细胞高于DC-CIK组(P<0.05);在5∶1、10∶1、20∶1效靶比范围内,培养14 d的DCCIK-Tregdel组细胞对SGC-7901胃癌细胞株的杀伤率明显高于DC-CIK组(P<0.05),并且这种杀伤作用随着效靶比的增高而增强;DC-CIK-Tregdel组培养上清液TGF-β水平低于DC-CIK组,而IFN-γ分泌水平明显高于DC-CIK组(P<0.05)。结论 CD4+CD25+T细胞对DC-CIK抗肿瘤效应细胞具有免疫抑制作用,去除CD4+CD25+T细胞能增加DC-CIK细胞增殖能力和抗肿瘤活性,为DC-CIK-Tregdel细胞有效治疗肿瘤提供了实验依据。
Objective The purpose was to investigate the effect of regulatory T cells on proliferation and kiUing activity of dendritic cells and cytokine-induced killer cells（DC-CIK） from peripheral blood. Methods DC-CIK ceils were prepared from peripheral blood mononuclear cells of healthy human. The experiment was divided into two groups： DC-CIK cells group（ conventional mononuclear cells were induced to DC-CIK cells） and DC-CIK-Tregael cells group （the peripheral mononuclear cells without CD4＋ CD25＋ regulatory T cell were induced to DC-CIK cells）. The proliferation ability and killing activity of the two groups cells were detected by cell expansion in multiples method and MTT assay;phenotype of regulatory T ceils were detected with flow cytometry. The TGF-β and IFN-γ were detected with ELISA in the cells culture supernatant of the two groups. Results The cell proliferation of the DC-CIK-Tregdel group cultured for 14 days was obvi- ously higher than that of the DC-CIK group（ P 〈 0.05 ）. Flow cytometry showed that the percentages of CD3＋ CD8＋ and CD3＋ CD56＋ T cells in DC-CIK-Tregdel group were significantly increased as compared with DC-CIK group（P 〈 0.05 ）. In the range of 5： 1,10： 1 and 20： 1 effeetor to target ratio,the killing rate of the DC-CIK-Tregdel group on gastric cancer cell line SGC-7901 was respectively increased than that of the DC-CIK group（ P 〈 0.05 ） and this killing effect was en- hanced with increased effector to target ratio. ELISA showed that the level of TGF-β in supernatant liquid of the DC-CIK- Tregdel group was lower than that of the DC-CIK group, while the level of IFN-γ in supernatant liquid of the DC-CIK-Tregde1 group was higher than that of the DC-CIK group （ P 〈 0. 05 ）. Conclusion CD4＋ CD25＋ T cells have immunosuppressive effect to anti-tumor activity of DC-CIK cells, and removing CD4＋ CD25＋ T cells can increase the DC- CIK cell proliferation and anti-tumor activity, which provides an experimental basis for the effective trea
出处
《中华全科医学》
2015年第3期349-352,共4页
Chinese Journal of General Practice
基金
山东省自然科学基金资助项目(ZR2013HL054)
