CIK的体外增殖及体内外杀瘤活性的实验研究

目的:从人骨髓造血前体细胞体外培养扩增树突状细胞(dendritic cells,DCs),测定其表型及T细胞刺激活性.方法:采用Mini-MACS分离技术,从正常人骨髓、脐血分离CD34~ 造血干细胞,体外以重组hGM-CSF,hTNF-α,hIL-3诱导培养2周,流式细胞术检测扩增细胞的表面表型及细胞内IL-12的表达,体外同种混合淋巴细胞反应检测扩增DCs的T细胞刺激活性.结果:从正常人骨髓、脐血分离得到高纯度(>90%)的CD34~ 造血干细胞,经重组hGM-CSF,hTNF-α的共同诱导培养,扩增得到大量DCs,加人hIL-3可以进一步增加DCs产量;FACS检测表明,扩增的DCs表达HLA-DR,CD40,CD54,CD80,CD86分子,细胞内有hIL-12的P35,P40亚基的表达;与外周血单核细胞培养生成的DCs相比,由CD34~ 干细胞扩增的DCs具有更强的激发同种T细胞增殖的能力.结论:人CD34~ 干细胞体外经诱导培养,可以生成大量功能成熟的DCs,从而为进一步开展DCs的基础及临床研究打下了基础.

The Proliferation Profile in vitro and Anti-Tumor Effects of CIK Cells in vivo and in vitro

Objective: To investigate the proliferation profile and the cytoloxicity of CIK (cytokine induced killer) cells, in vitro, and their anti-tumor effects on S180 bearing mice in vitro. Methods: CIK cells were generated by culturing PBMC in the presence of r-IFN on day 0 and IL-2, anti-CD3 McAb, IL-1 on day 1; CIK cultures were analyzed on different time points by FACS; Compared with LAK cells, the cytotoxicity in vitro by MTT assays and the anti-tumor effects on S180 bearing mice in vivo of CIK cells were tested respectively. Results: The flow cytometry analysis showed that CD3 CD56 T cells which were the major cytolytic effectors in CIK cells expanded 1 000-fold after 2 weeks in CIK culture ; CIK cells had greater cytotoxicity against tumor cells in vitro than that of LAK cells; and had also significantly anti-tumor effects in mice bearing S180 tumor compared with LAK cells in vivo. (median inhibitory rates 78% vs 33%, P < 0.05 t ); The CIK therapy was possibly correlated with induction of the activation of the splenic T lymphocytes of S180 bearing mice. Conclusion: CIK cells were highly efficient cytolytic effector cells which were non-MHC restricted