负载肝癌抗原的树突状细胞瘤苗诱导的抗肿瘤作用

Antitumor Effects Induced by Dendritic Cells Pulsed with Hepatocarcinoma Cells Antigen

目的探讨原发性肝癌患者外周血树突状细胞(DC)体外经自体肝癌细胞抗原致敏后诱导的抗肿瘤作用。方法肝癌患者外周血经梯度密度离心法分离,获得DC前体细胞,用重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素-4(rhIL-4)联合培养,诱导扩增DC。制备自体肝癌细胞抗原,体外脉冲DC,检测DC诱导自体T细胞增殖能力及细胞毒性T细胞(CTL)在体外对自体肝癌细胞的杀伤活性,并检测肿瘤抗原致敏DC分泌的IL-1 2水平。结果经自体肝癌细胞抗原致敏的DC能分泌IL-1 2和诱导较强的自体T细胞增殖,且能诱导特异性CTL,该CTL对自体肝癌细胞具有很强的杀伤活性,杀伤率明显高于DC、未经肝癌细胞抗原致敏的DC激活的CTL及T淋巴细胞的杀伤率,而对3 LL LEWIS肺癌细胞、H 2 2肝癌细胞则无明显的杀伤作用。结论肝癌患者外周血DC经自体肝癌细胞抗原致敏后能诱导高效而特异的抗肝癌免疫,其机制可能与增强T细胞应答和诱导机体产生肿瘤特异CTL而发挥特异性的抗肿瘤作用有关。

Objective：To investigate the antitumor effects induced by dendritic ceils pulsed with hepatocarcinoma cells antigen. Methods： DCs were propagated from peripheral blood of hepatocarcinoma patients （n= 5） in vitro with rhGM-CSF and rhlL-4. The autogenous hepatocarcinoma cells antigen was used to activate DCs. The T cell stimulating activity of DC was detected by 3H-TdR labbel assay and the cytotoxicity of CTL was assayed by the lactate dehydrogenase （LDH） release assay. The level of IL-12 in the superntant was detected by ELISA. Results： The autogenous hepatocarcinoma cells antigen pulsed DCs could secrete IL-12 and effectively stimulate syngeneic T lymphocytes to proliferate and could effectively stimulate the responsiveness of syngeneic T cells to induce specific CTL and the killing activity against autogenous hepatocarcinoma cells in vitro was higher strikingly than those of controls,but no significant killing activity on 3LL LEWIS ceils or H22 ceils was observed. Conclusion： DC from hepatocarcinoma patients could induce efficient and special immune response against hepatocarcinoma. Its antitumor immunity mechanism might be that DC pulsed with hepatocarcinoma cells antigen enchaneed T lymphocytes proliferation and induced tumor specific CTL to elicit specific antitumor effects.