唑来膦酸刺激健康人和骨肉瘤PBMCs体外扩增γδT细胞

Amplification of γδ T cells in PBMCs of healthy donors and osteosarcoma patients stimulated by zoledronate

目的:探讨唑来膦酸(Zol)联合IL-2对健康人和骨肉瘤患者末梢血γδT细胞的扩增和细胞毒性作用的影响。方法:取健康人和骨肉瘤患者外周血来源的单个核细胞,分别使用IL-2和Zol联合IL-2刺激体外扩增,流式细胞技术检测γδT细胞的纯度,并计算其扩增倍数。以Daudi细胞作为阳性对照、Raji细胞为阴性对照,并以人成骨细胞系hFOB细胞作为正常对照,用LDH法检测扩增后γδT细胞的细胞毒活性的变化。结果:健康人和骨肉瘤患者外周血单核细胞经过体外14 d的培养,Zol联合IL-2组可高度选择性扩增γδT细胞,并且扩增后γδT细胞具有杀伤活性,而对正常细胞无杀伤活性。结论:健康人和骨肉瘤患者PBMCs经唑来膦酸联合IL-2刺激后,可获得高纯度具有较强的杀伤作用的γδT细胞。

AIM： To investigate the amplification and cytotoxicity of γδ T cells in peripheral blood mononuclear cells（PBMCs） of healthy donors and osteosarcoma patients stimulated by zoledronate（Zol） and IL-2.METHODS： PBMCs from healthy donors and osteosarcoma patients were stimulated with IL-2 and Zol＋IL-2,respectively.After 14-day culture,the purity of γδ T cells was assessed by flow cytometry.The cytotoxicity of γδ T cells against target cells was analyzed using a standard lactate dehydrogenase release assay with γδ T lymphocyte-sensitive Daudi cells,γδ T lymphocyte-resistant Raji cells and human osteoblast cell line,hFOB,as the target cells.RESULTS： After 2-week culture ex vivo of PBMCs from healthy donors and osteosarcoma patients,compared with stimulation of IL-2,Zol＋IL-2 significantly promoted the amplification of γδ T cells.In addition,γδ T cells showed the higher cytotoxicity against Daudi cells,but no cytotoxic effect on normal cells like hFOB.CONCLUSION： γδ T cells of high purity and high cytotoxicity can be obtained by the stimulation of Zol combined with IL-2 on PBMCs from healthy donors and osteosarcoma patients.