肝癌细胞裂解物致敏的树突状细胞瘤苗体外诱导特异性抗肝癌免疫

An in vitro study of specific antitumor immunity induced by dendritic cells pulsed with tumor cell lysates from patients with hepatocellular carcinoma

目的 探讨肝癌患者肿瘤细胞裂解物致敏的树突状细胞(DC)瘤苗体外诱导自体T淋巴细胞特异性抗肝癌免疫效应。 方法 从肝癌患者外周血单个核细胞中诱导D C,用重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和白细胞介素-4(rhIL-4)刺激活化,经自体肝癌细胞裂解物致敏。用流式细胞仪检测D C细胞表面分子的表达,酶联免疫吸附法检测T淋巴细胞培养上清液中干扰素(I F N)γ和白细胞介索-12(IL-12)的含量,液体闪烁计数仪测定肝癌细胞裂解物致敏的D C刺激自体T淋巴细胞增殖效应,四甲基偶氮唑盐法检测肝癌细胞裂解物致敏D C诱导的细胞毒T淋巴细胞对自体肝癌细胞的特异性杀伤作用。 结果 肝癌细胞裂解物致敏的DC瘤苗可上调DC表面CD1 a、CD40、CD86和人类白细胞抗原-DR分子表达水平,其与T淋巴细胞共培养产生的IFN γ、IL-12的浓度明显高于未致敏的D C组(t值分别为2.30、2.14,P<0.05),肝癌细胞裂解物组(t值分别为14.01、15.40,P<0.01)和对照组(t值分别为14.85、16.87,P<0.01)。同时肝癌细胞裂解物致敏的瘤苗可明显诱导T淋巴细胞的增殖,其诱导的细胞毒性T淋巴细胞对自体肝癌细胞的杀伤率(81.72％±9.49％)显著高于对HepG2的杀伤率(49.37％±11.21％)和人鼻咽癌肿瘤细胞的杀伤率(17.14％±5.65％),P<0.01。 结论 肝癌细胞?

Objective To investigate T cell-mediated antitumor effects of dendritic cells (DCs) pulsed with tumor lysates in patients with hepatocellular carcinoma (HCC) in vitro. Method DCs isolated from peripheral blood mononuclear cells of HCC patients were cultured and proliferated in vitro by using recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4), and then were pulsed with autologous hepatoma cell lysates. The phenotypes of the DCs were assayed by flow cytometry. The concentration of IFN γ and IL-12 were measured in culture supernatants by ELISA. The ability of DCs pulsed with hepatoma cell lysates to stimulate proliferation of autologous T lymphocytes (CTL) was tested by thymidine incorporation method. The specific cytolytic activity of CTL was assessed by MTT method. Results The hepatoma cell lysates pulsed DC vaccines led to up-regulation of CDla, CD40, CD86 and HLA-DR. Concentrations of IFN γ and IL-12 were increased more in the hepatoma cell lysate pulsed DCs group than those in the impulsed DCs group (P < 0.05), the hepatoma cell lysate group and control group (P < 0.01). The proliferation of T-cells was markedly enhanced in the hepatoma cell lysate pulsed DCs group than that in the others (P < 0.01). The CTL stimulated by the hepatoma cell lysate pulsed DCs had much higher cytotoxicity to autologous hepatoma cells (killing rate: 81.72% ± 9.49%), as compared with HepG2 and HNE-1 tumor cells (killing rate: 49.37% ± 11.21% and 17.14% ± 5.65%, respectively), (P < 0.01). Conclusions The hepatomacell lysate pulsed DC vaccines can induce an effective and specific anti-hepatoma effect.