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目的探讨转染肝癌细胞总RNA转染的树突状细胞(DC)疫苗抑制肝癌作用及其机制。方法采用粒/巨噬细胞集落刺激因子(GM-CSF)和白细胞介素(IL)-4联合培养6 d成小鼠骨髓来源不成熟DC(iDC),质脂体转染肝癌细胞系Hepa1-6 RNA入iDC,用LPS刺激后成为Hepal-6 RNA/DC疫苗,对照组为iDC组、LPS/DC组Saline组,以每只2×10~6个细胞腹腔注射免疫小鼠每周1次共3次,然后接种5×10~5 Hepal-6细胞,观察肿瘤生长情况、特异性CTL活性的测定以及抗体阻断实验。结果实验组Hepal-6 RNA/DC组肿瘤在第8周为(8．5±3．6)mm,而iDC组、LPS/ DC组、Saline对照组分别为(36．6±3．6)、(31．3±4．7)、(32．2±4．0)mm,与实验组比较差异均有统计学意义(P＜0．05)。实验组脾细胞对Hepal-6细胞有特异性杀伤效应,而对肺癌LLC细胞无杀伤活性。所诱导的抗肿瘤效应细胞包括CD~(8+)、CD~(4+)T淋巴细胞。结论肝癌细胞的总RNA转染的DC肿瘤疫苗能诱导CD~(8+)、CD~(4+)T细胞免疫,是较有临床应用前景的肝癌免疫治疗方法。
Objective To investigate the inhibitory effect of dendritic cell (DC) vaccine transfected with total RNA of hepatoma cells on hepatocellular carcinoma and its mechanism. Methods Mice bone marrow-derived immature DCs (iDC) were cultured with GM-CSF and IL-4 for 6 days. The liposomes were transfected into Hepa1- 6 RNA was injected into iDC and HepPS-6 RNA / DC vaccine was stimulated with LPS. The control group was iDC group and LPS / DC Saline group. The mice were immunized intraperitoneally with 2 × 10 ~ 6 cells once a week A total of three times, then inoculated 5 × 10 ~ 5 Hepal-6 cells, observed tumor growth, specific CTL activity and antibody blocking experiments. Results The tumor volume of Hepal-6 RNA / DC group was (8.5 ± 3.6) mm in the experimental group and (36.6 ± 3.6) in the iDC group, LPS / DC group and Saline control group , (31.3 ± 4.7) and (32.2 ± 4.0) mm, respectively, which were significantly different from the experimental group (P <0.05). The experimental group of spleen cells have a specific killing effect on Hepal-6 cells, but no cytotoxic activity on lung cancer LLC cells. Induced antitumor effector cells include CD ~ (8 +) and CD ~ (4 +) T lymphocytes. CONCLUSION: DC tumor vaccine transfected with total RNA of hepatoma cells can induce CD8 +, CD4 + T cell immunity and is a promising immunotherapy for hepatocellular carcinoma.