树突状细胞介导的T淋巴细胞对肝癌细胞的特异性杀伤效应

目的通过体外诱导高效特异的抗肝癌细胞系免疫反应,制备一种以树突状细胞(DC)为基础的肝癌疫苗。方法分离人外周血单核细胞,经IL4和GMCSF联合刺激诱导DC分化。培养第4天,DC用人肝癌细胞系SMMC7721的冻融抗原致敏。培养第7天,将肝癌冻融抗原致敏的DC和未经肝癌冻融抗原致敏的DC分别与T淋巴细胞混合培养24h作为效应细胞,以人SMMC7721肝癌细胞、MCF7乳腺癌细胞和PC3前列腺癌细胞作为靶细胞,用MTT法检测细胞毒活性。结果经肝癌冻融抗原致敏的DC对SMMC7721细胞有高效而特异的杀伤活性,对其他肿瘤细胞系也有一定的杀伤活性,但无明显特异性;未经肝癌冻融抗原致敏的DC对3种肿瘤细胞系均有一定的杀伤活性,但无明显特异性;各组效应细胞对靶细胞的杀伤率随效靶比(EC/TC)的升高而增强。结论以肝癌细胞冻融抗原冲击致敏的DC刺激T淋巴细胞增殖在体外可诱导产生高效特异性的抗肝癌免疫反应,为肝癌的主动特异性免疫治疗提供了可靠的理论依据。

Specific cytotoxic effects mediated by dendritic cells on liver carcerous cells in vitro

Objective To induce highly effective and specific immunologic reaction specific to hepatocarcinoma cell line, with the aim of producing anti-hepatocarcinoma vaccine based on dendritic cells. Methods Separating monocytes from human peripheral blood, inducing DC differentiation in the presence of GM-CSF and IL-4 in vitro. On the fourth day of culture, DC was sensitized with SMMC-7721 hepatocarcinoma cell lysate antigen. On the seventh day, DC and lysate antigen pulsed-DC were cocultured with T lymphocyte for 24h, respectively, both of which were effector cells. SMMC-7721 cells, MCF-7 breast carcinoma cells and PC-3 prostate carcinoma cells were used as target cells. Cytotoxic rates were detected using MTT assay. Results The cytotoxic rate of lysate antigen pulsed-DC to SMMC-7721 cells was highly effective and specific, while that to MCF-7 cells and PC-3 cells were effective to some extent without obvious specificity. The cytotoxic rates of non-pulsed DC to SMMC-7721 cells, MCF-7 cells and PC-3 cells were also effective, to some extent, without obvious specificity. The cytotoxic rate of effector cells to target cells increased with proportion of EC to TC. Conclusion DC pulsed by hepatocarcinoma cells lysate antigen can stimulate proliferation of T lymphocyte and induce highly effective and specific anti-hepatocarcinoma immunity. This study lay a sound foundation for next study in vivo and clinical experiment, find out a very promising way for active specific immuotherapy of primary hepatic carcinoma.