多种细胞因子诱导的杀伤细胞体外扩增及对骨肉瘤细胞杀伤活性的研究

目的探讨不同人群来源的多种细胞因子诱导的杀伤细胞（cytokine—induced killer cells，CIK）细胞增殖及细胞毒活性的作用，为CIK细胞在骨肉瘤中的过继免疫治疗提供实验依据。方法通过向健康人及骨肉瘤患者各10名男性的外周血单核细胞（PBMC）中加入IL-2、IFN-γ、IL-1及抗CD3McAb，诱导出CIK细胞，用光镜、倒置显微镜观察细胞形态，细胞记数板法计数细胞，四甲基偶氮哇盐（MAW）法检测收获细胞对相应骨肉瘤细胞的细胞毒作用。结果二组均能成功诱导出CIK细胞，其中健康人组体外增殖能力、细胞毒作用均优于骨肉瘤患者组，各组CIK细胞随效靶浓度比的递增其杀瘤活性相应增加（P〈0．05），二组CIK细胞对骨肉瘤细胞48h杀伤率明显高于24h（P〈0．05）。结论CIK细胞的体外增殖力及对骨肉瘤细胞的杀伤活性与机体状态有关，CIK细胞对骨肉瘤细胞杀伤活性存在量效及时效关系。

A study on the proliferation and cytotoxicity of cytokine-induced killer cells

Objective To study various activation on proliferation and cytotoxicity of CIK cells and provide advanced evidence of the immunotherapy to ostesarcoma. Methods Separate PBMC from peripheral blood of 10 male and 10 men with ostesarcoma, induced to CIK cells by supplementing cytokines of IL-2, IFN-γ, IL-1 and anti- CD3McAb in PBMC. The morphology of CIK cells were detected by using microscope and invert microscope. CIK cells proliferation is tested by blood cell recording board. The killing ostesarcoma cell activity of CIK cells were tested with MTT method. Results Both groups were successfully induced to CIK cells. The proliferation rate and cytotoxicity of the normal adults-CIK were stronger than the ostesarcoma patient-CIK cells. Each group of CIK cells cytotoxicity increased progressively along with increasing of the effect target density ratio （P〈0.05）.48h killing rat of CIK cells obviously were higher than 24h in two group （P〈0.05）. Conclusions CIK proliferation and cytotoxicity in vitro are related to organism condition. CIK cytotoxicity exist quantity effect and effectiveness of time relations.