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目的研究脐血单个核细胞体外多向诱导分化为LAK,CD3AK,CIK细胞与体外对胃癌细胞株MGC-803的杀伤活性。方法采用IL-2刺激诱生LAK细胞,用CD3单抗和IL-2刺激诱生扩增CD3AK细胞,用加入IFN-r,24h后加入IL-1,CD3单抗,IL-2刺激诱生CIK细胞,观察它们的扩增情况,并分析其相互关系;取培养的CIK细胞用流式细胞仪分析表型;用MTT、法以MGC-803胃癌细胞株为靶细胞,分组测定LAK,CD3AK,CIK细胞对瘤细胞的杀伤活性。结果CD3AK和CIK细胞的扩增能力远大于IAK细胞(P<0．05)。CIK细胞和CD3AK相比,在前期无明显差异,至第20天左右时CIK细胞的扩增能力显著高于CD3AK的扩增倍数(P<0．05)。流式细胞仪分析结果显示CIK细胞培养后,其主要效应细胞CD3+CD56+细胞较培养前显著增加(P<0．05),CD8+细胞也较培养前显著增加(P<0．05)。CD3AK和CIK细胞对MGC-803胃癌细胞株的杀伤活性均显著高于LAK细胞(P<0．05),而CIK细胞杀伤活性强于CD3AK细胞(P<0．05)。结论CIK细胞具有更高的扩增能力和更强的杀伤活性,是肿瘤过继免疫治疗中更为有效的杀伤效应细胞。
OBJECTIVE To study the multidrug induced differentiation of cord blood mononuclear cells into LAK, CD3AK and CIK cells in vitro and their cytotoxicity to gastric cancer cell line MGC-803 in vitro. Methods LAK cells were induced by IL-2 stimulation, CD3AK cells were stimulated with CD3 monoclonal antibody and IL-2, IFN-γ was added and IL-1, CD3 and IL-2 were stimulated by IL-2 stimulation CIK cells to observe their amplification, and to analyze their correlation; take cultured CIK cells by flow cytometry analysis of phenotype; using MTT method to MGC-803 gastric cancer cell lines as target cells, the group measured LAK, CD3AK , CIK cells on tumor cell killing activity. Results The amplification of CD3AK and CIK cells was much larger than that of IAK cells (P <0.05). Compared with CD3AK, there was no significant difference between CIK cells and CD3AK cells. The proliferation ability of CIK cells at day 20 was significantly higher than that of CD3AK (P <0.05). Flow cytometry analysis showed that after CIK cells were cultured, the main effector cells CD3 + CD56 + cells were significantly increased (P <0.05) and CD8 + cells were significantly increased (P <0.05). The cytotoxicity of CD3AK and CIK cells to MGC-803 gastric cancer cell lines was significantly higher than that of LAK cells (P <0.05), while the cytotoxic activity of CIK cells was stronger than that of CD3AK cells (P <0.05). Conclusion CIK cells have higher amplification ability and stronger killing activity, and are more effective killer effector cells in adoptive immunotherapy of tumor.