细胞因子诱导的杀伤细胞基本特性及抗肿瘤效应研究

The study on the basic characteristics and the anti-tumor effects of CIK cells

目的建立培养细胞因子诱导的杀伤(CIK)细胞方法,并评价其基本特点、体内外抗肿瘤活性及安全性。方法使用抗CD3单克隆抗体、rhIL-2及rhIFN-γ刺激PBMC细胞,使其在体外大量扩增,利用流式细胞术、体外杀伤实验及荷人卵巢癌小鼠模型检测CIK细胞的特点及抗肿瘤活性;并检测CIK细胞中外源性因子污染情况及其急性毒性作用。结果体外培养21 d后,CIK细胞总数可达1010以上,其主要成分为CD3+CD8+T细胞及CD3+CD56+NKT细胞,CIK细胞在体外对Ho8910、A549、K562及HepG2细胞均有较高的杀伤活性,并对荷瘤小鼠具有治疗作用。同时在CIK细胞中未检测到外源性病原体污染。结论 CIK细胞在体内外均具有较高的抗肿瘤活性且无毒性作用。

Objective To establish the methods of cytokine-induced killer （CIK）cells and evaluate the characteristics,anti-tumor activity in vitro and in vivo and the safety.Methods Anti-CD3 mAb,recombinant human interleukin 2 （rhIL-2 ）and recombinant human interferon-γ（rhIFN-γ）were used to induce the peripheral blood mononuclear cells （PBMCs）proliferation abundantly in vitro.Flow cytometry assay （FACS）,in vitro cytotoxicity as-say and tumor mouse model xenografted with Ho-8910 （human ovarian cancer cell line）were used to detect the char-acteristics and the anti-tumor activityof CIK cells.Additionally,the contamination of exogenous agents and the acute toxicity effect of CIK cells were also determined.Results After 21 days in vitro culture,the total number of CIK cells were more than 10^10,among which there were mainly CD3^＋CD8^＋T cells and CD3^＋CD56^＋NKT cells.CIK cells dis-played high cytotoxicity against Ho8910,A549,K562 and HepG2 cells in vitro,and they also showed therapeutic effects in tumor xenografted mice.Additionally,no acute toxicity was observed in CIK cells-treated mice,and there was also no contamination of bacteria,fungus,mycoplasma and virus in CIK cells.Conclusion CIK cells have high anti-tumor activity without any toxic side effects.