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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 the effect of dendritic cell (DC) -sensitized dendritic cells (DCs) sensitized by tumor cell lysates to the induction of autologous T lymphocyte-specific anti-HCC in vitro. Methods DCs were induced from peripheral blood mononuclear cells of patients with hepatocellular carcinoma and activated by recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) and interleukin-4 (rhIL-4) Sensitization. Flow cytometry was used to detect the expression of DC cell surface molecules. The levels of IFN-γ and IL-12 in T lymphocyte culture supernatants were detected by enzyme-linked immunosorbent assay. The proliferation of autologous T lymphocytes was stimulated by DCs sensitized by liver cancer cell lysate and the specific killing effect of DC induced cytotoxic T lymphocytes on autologous hepatocarcinoma cells was detected by MTT assay . Results Liver cancer cell lysate-sensitized DCs could up-regulate the expression of CD1a, CD40, CD86 and HLA-DR on the surface of DCs. The concentrations of IFN-γ and IL-12 produced by co-culture with T lymphocytes were significantly higher In untreated DC group (t = 2.30, 2.14, P <0.05), liver cancer cell lysate group (t = 14.01,15.40, P <0.01) and control group (t = 14.85,16.87 , P <0.01). At the same time, the hepatoma cell lysate-sensitized tumor cells could obviously induce the proliferation of T lymphocytes, the cytotoxic T lymphocyte-induced killing rate of autologous hepatoma cells (81.72% ± 9.49%) was significantly higher than that of HepG2 cells 49.37% ± 11.21%) and the killing rate of human nasopharyngeal carcinoma cells (17.14% ± 5.65%), P <0.01. Conclusion Liver cancer cells?