DCCIK细胞生物学活性及抗淋巴瘤细胞作用.docVIP

DCCIK细胞生物学活性及抗淋巴瘤细胞作用 　 作者：魏绪仓，连小云，赵文理，杨娣娣，韩秀蕊，邢佩霓，李梅生 【摘要】 目的 研究 细胞因子诱导的杀伤细胞(CIK)与树突状细胞(DC)共培养后DCCIK细胞的体外增殖能力、免疫表型变化、分泌细胞因子水平以及抗淋巴瘤细胞活性。 方法 正常人外周血单个核细胞诱导DC和CIK细胞，将DC与CIK共培养，以CIK细胞单独培养为对照。用台盼蓝活细胞计数 计算 细胞扩增倍数，MTT法测定杀伤活性，流式细胞术 分析 免疫表型，ELISA双抗体夹心法检测分泌干扰素γ(IFNγ)、白细胞介素12(IL12)的水平。结果 DCCIK细胞增殖能力明显高于CIK细胞(Plt;0.05)；DC、CIK细胞共培养后，CD3+CD8+、CD3+CD56+双阳性细胞比率较同条件下CIK细胞组显著增多(Plt;0.05)；共培养3d，DCCIK细胞上清液中IL12、INFγ的分泌量均比CIK细胞单独培养的分泌量高(Plt;0.01，Plt;0.05)；在5∶1-40∶1的效靶比范围内，DCCIK细胞对淋巴瘤细胞的杀伤率显著高于CIK细胞(Plt;0.05)，且杀伤率与效靶比呈正相关。结论 DCCIK细胞的增殖能力、分泌细胞因子水平、抗淋巴瘤细胞活性均高于CIK细胞，为DCCIK细胞免疫 治疗 提供了实验和 理论 依据。 【关键词】 CIK细胞；DCCIK细胞；生物学活性；抗淋巴瘤 论文代写 　　 The biological activity and antitumor effect against　lymphoma cells in DCCIK cells 　　ABSTRACT: Objective To investigate the proliferation activities, phenotype changes, level of secretory cytokines and antitumor activity against lymphoma cells of DCCIK cells in coculture of cytokineinduced killer (CIK) cells with dendritic cells (DC). Methods DC and CIK were prepared from healthy human peripheral blood mononuclear cells. They were cocultured peripherally. CIK cells were cultured alone as controls. Increased number of cells were counted by tapanblue staining, killing activity was detected by MTT assay, cells phenotypes were analyzed by flow cytometry, secretions of INFγ and IL12 were determined by ELISA. Results The proliferation activity of DCCIK cells was significantly higher than that of CIK cells (Plt;0.05). Under the same condition, the ratio of CD3+ CD56+ and CD3+ CD8+ double positive cells in CIK cells was significantly enhanced by coculture with DC (Plt;0.05). In DCCIK cells, the level of IL12 and INFγ in culture supernatants was increased noticeably on day 3 compared to CIK cells which were cultured alone (Plt;0.01, Plt;0.05). At the effectortarget ratio from 5∶1 to 40∶1, the antitumor effect of DCCIK cells was much higher than that of CIK cells (Plt;0.05), and this effect was positively related with the eff