健康人和肝癌患者CIK体外增殖能力及对原代肝癌细胞抗肿瘤作用的比较研究

目的探讨源自健康人和肿瘤患者的细胞因子诱导的杀伤细胞(CIK)体外增殖能力及对原代肝癌细胞的抗肿瘤作用的差异,进一步了解CIK的抗肿瘤作用.方法分离健康人和肝癌患者外周血单个核细胞经细胞因子激活诱导培养为CIK,流式细胞分析检测CIK免疫表型;用机械研磨法将新鲜肝癌组织标本分离成单细胞悬液;四甲基偶氮唑盐法榆测两种CI...

Analysis of the proliferation ability and the antineoplastic activity of CIK cells against original hepatocellullar cancer cells between healthy adults and tumor patients

Objective To explore the differences of the proliferation ability and the antineoplastic activity of CIK cells against origenal hepatocellullar cancer cells between healthy adults and tumor patients in vitro. Methods Peripheral blood mononuclear cells (PBMC) from healthy donors and tumor patients were incubated to induce CIK cells in the presence of interferon gamroa(IFN-γ) ,IL-2 and anti-CD3 monoclonal antibody (mAb). The changes in the proliferation activity and phenotypes of the CIK cells were identified by flow cytometric analysis. Single cell suspension was prepared bom the fresh hepatocellular carcinoma tissue by using mechanical trituration method. MTT assays were used to determine the cytotoxicity of CIK cells against origenal hepatocellullar cancer cells. Results The CIK cells from both healthy donors and tumor patients were significantly increased with the extension of time. The expression rate of CD3+/CD56+ cells from healthy donors rose from 1.053% ±0.22% on 1st day ,25.36% ±2.19% on the 7th day to 55.12% ±1.99% on the 14th day ,which was significantly higher than that of tumor patients ( P ＜ 0.05). MTT assays showed that the cytotoxicity of CIK cells from both enhanced obviously with the addition of Effect/Target rate and extension of time(P ＜0.05). After the origenal hepatocellullar cancer cells were treated by CIK cells 24 hours, the cytotoxicity of CIK cells from healthy donors at the effector:target ratio of 10 ∶ 1,20 ∶1 and 40 ∶1 was significantly higher than that of tumor patients(P＜0.05) ,respectively. After treated 48 hours, compared with tumor patients, the cytotoxicity of healthy donors? CIK at the three effector:target ratio was alsosignificantly higher(P ＜0.05), respectively. Conclusion CIK cells from both have amplification ability and cytotoxic activity in vitro,and the proliferation ability and killing activity of healthy adults CIK is stronger than that of tumor patients,which provides an experimental basis for CIK to clinical application as an adoptive immunotherapy.