舌癌患者CIK细胞的体外培养及其免疫活性

The cell culture in vitro and the immunoe characteristics of CIK from tongue cancer patients

目的研究舌癌患者外周血CIK细胞的增殖能力、细胞表型和细胞毒作用，寻求对舌癌过继免疫治疗的新途径。方法提取20例舌癌患者和20例健康人外周血单核细胞用IFN-孔抗CD3mAb、IL-2和IL-1β共同培养诱导CIK细胞，流式细胞仪检测细胞表型特征，用MTT法检测细胞对Tca8113的杀伤活性；同时培养舌癌患者LAK细胞、CD3AK细胞和舌癌患者CIK细胞在增殖和杀瘤活性上作比较。结果舌癌患者20ml外周血经过一个周期培养平均获得的CIK细胞数低于健康对照组（P〈0．05），但舌癌患者CIK细胞经过周期培养也较培养前的数量明显增高，于12～21d达高峰。舌癌患者CIK细胞与LAK相比，增殖倍数和杀伤活性均明显提高（P〈0．05）；舌癌患者CIK增殖倍数与CD3AK相比，早期差异无显著性，但后期增殖倍数及杀伤活性有显著提高（P〈0．05）。结论舌癌患者CIK细胞体外增殖倍数和杀瘤活性明显优于LAK和CD3AK细胞，低于正常人CIK的增殖能力，而舌癌患者CIK细胞杀伤活性与正常人来源CIK细胞差异无显著性（P〉0．05），有望作为自体过继免疫治疗舌癌的一种新方法。

Objective To study the proliferation, cell phenotype, cytotoxicity of CIK cell from peripheral blood of tongue cancer patients. Explore the novel immunotherapy of tongue cancer. Methods Peripheral blood mononuclear cell（PBMC） were isolated from 20 cases of tongue cancer patients and 20 healthy donors. Expansion of CIK cells by the timed addition of interferon gamma （ IFN-γ）, CD3 monoclonal antibody （ CD3mAb）, interleukin-2 （ IL- 2） and interleukin-β（IL-1β）. Cell phenotype was assayed by FCM. Cytotoxicity was assayed by MTT and CIK cells were compared by LAK cell, CD3AK cell in the same time. Results The number of CIK from tongue cancer patients was lower than that from healthy donors. The multiplication ability of CD3AK and CIK cells was higher than that of LAK and CD3AK cells markedly（P 〈0. 05）. The killing activity of CIK cells was higher than that of LAK and CD3AK cells markedly （ P 〈 0.05 ）. Conclusion The multiplication ability or cytotoxicity of CIK cells from tongue cancer patients is higher than those of LAK cells and CD3AKs. It is another more efficient killer cell in the therapy of tongue cancer adoptive immunity.