目的:探讨hKDR融合基因致敏树突状细胞(DC)激发的细胞毒性T淋已细胞(CTL)对小鼠肝癌细胞的杀伤活性。方法:自小鼠骨髓中获取DC,以粒细胞-巨噬细胞集落刺激因子(GM-CSF)和hKDR融合基因的mRNA致敏DC;用致敏的DC免疫小鼠取其脾脏获得CTL,即效应细胞,用脂质体将pCDNA hKDR转染至小鼠肝癌Hepal-6细胞株中,作为靶细胞;将效应细胞与靶细胞按不同比例混合,用乳酸脱氢酶(LDH)释放法检测细胞毒作用。结果:未经修饰的DC激活的CTL与靶细胞比例为100:1、50:1、25:1情况下,其杀伤率分别为19.2%、12.3%、6.9%;经hKDR修饰的DC激活的CTL与靶细胞比例为100:1、50:1、25:1情况下,其杀伤率分别为71.6%、55.8%、22.7%,将两组结果在相同的比例下两两比较,均有显著性差异(P〈0.05)。结论:hKDR融合基因致敏DC激活的CTL对小鼠肝癌细胞具有较强的杀伤作用,且杀伤能力随效应细胞与靶细胞比例增高而增强。
Objective:To investigate the lethal effect of cytotoxic T lymphocyte (CTL) stimulated by dendritic cells (DC) modified by hKDR fusion gene on hepatoma carcinoma cells in mice. Methods:Bone marrow precursor-derlved dendritic cells (BMDCs) were harvested from bone marrow and sensitized with rmGM-CSF and hKDR-mRNA. Mice were immunized with the sensitized DCs to produce CTL, the effector cells, pCDNA hKDR was transfected with liposome to Hepal-6 cell,a routine hepatoma carcinoma cell line,serving as target cell. The effector cells and target cells were mixed in various proportions, And Cyto-Tox 96a Non-Radioactive Cytotoxicity Assay was performed to determine cytotoxic effect. Results:In the ratios of 100: 1,50: 1 ,25: 1 ,the kill rates of CTL activated by non-modified DC were 19.2% ,12.3% and 6. 9% ,respectively,whereas the kill rates of DC activated by hKDR fusion gene were 71,6% ,55.8% and 22,7% in the same mixture proportions. There was a significant difference between the non-modlfied and modified two groups in each proportion (P〈0.05). Conclusions: It was shown that CTL induced by DC plused with hKDR fusion gene had lethal effect on mouse hepatoma carcinoma cell in vitro, and this effect got enhanced as the proportion of effcctor cells became increased,