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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 anti-tumor effect of dendritic cells (DCs) dendrites induced by autologous hepatocarcinoma cells in patients with primary liver cancer. Methods Peripheral blood of patients with hepatocellular carcinoma was isolated by gradient density centrifugation. DC precursor cells were obtained and cultured with recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and recombinant human interleukin-4 (rhIL-4) , Induce the expansion of DC. To prepare auto-hepatoma cell antigen, pulsed DC in vitro, detect the ability of DC to induce autologous T cell proliferation and cytotoxic T lymphocyte (CTL) cytotoxicity to autologous hepatocarcinoma cells in vitro, and detect the level of IL-12 secreted by tumor antigen-primed DC . Results DCs sensitized with autologous hepatocarcinoma cells could secrete IL-12 and induce stronger autologous T cell proliferation, and induce specific CTL. The CTL has strong cytotoxic activity on autologous hepatocarcinoma cells with high killing rate The cytotoxicity of CTL and T lymphocytes activated by DCs that were not sensitized by liver cancer cell antigen was not obvious in 3 LL LEWIS lung cancer cells and H 2 2 liver cancer cells. CONCLUSION: DCs induced by autologous hepatocarcinoma cells can induce efficient and specific anti-hepatocellular immunity after being sensitized by autologous hepatocarcinoma cells. The mechanism may be related to the enhancement of T cell responses and the induction of tumor-specific CTLs in the body to exert specific antitumor effects.