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目的研究粒细胞巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)分泌型肝癌疫苗对移植性肝癌小鼠细胞毒性T淋巴细胞杀伤活性的作用机制。方法本实验建立3个组别:肝癌疫苗组(A组),肝癌组(B组)及PBS对照组(C组)。在小鼠体内注入H 22肝癌细胞构建移植性肝癌组,同时建立GM-CSF分泌型肝癌疫苗组及PBS对照组;采用流式细胞术检测每组小鼠外周血中CD8+T细胞免疫效应分子的表达水平;采用MTT方法检测各组脾细胞细胞毒T淋巴细胞(CTL)的杀伤活性;采用Western blot方法检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和γ-干扰素(γ-interferon,γ-INF)的表达水平。结果流式细胞术结果表明,与肝癌组相比,GM-CSF分泌型肝癌疫苗可显著地增加小鼠外周血中CD8+T细胞的表达(P<0.01);MTT结果表明,与肝癌组相比,GM-CSF分泌型肝癌疫苗可显著地增加脾细胞细胞毒T淋巴细胞的杀伤活性(P<0.01);Western blot结果表明,与肝癌组相比,GM-CSF分泌型肝癌疫苗组可显著下调TNF-α和γ-INF蛋白的表达(P<0.01)。结论 GM-CSF分泌型肝癌细胞疫苗可显著地抑制H22肝癌细胞H22的活性,其作用机制可能是通过活化CD8+T细胞的表达和提高脾细胞细胞毒T淋巴细胞的杀伤活性,降低TNF-α和γ-INF蛋白的表达。
Objective To study the mechanism of granulocyte-macrophage colony stimulating factor (GM-CSF) secreting hepatocarcinoma vaccine on the cytotoxic T lymphocyte cytotoxicity in mice with transplanted hepatoma. Methods Three groups were established in this study: Hepatocarcinoma vaccine group (A group), hepatocellular carcinoma group (B group) and PBS control group (C group). The H22 hepatoma cells were injected into mice to establish the transplanted liver cancer group, and the GM-CSF secreting hepatocarcinoma vaccine group and the PBS control group were also established. The immune effector molecules of CD8 + T cells in peripheral blood of each group were detected by flow cytometry The cytotoxic activity of cytotoxic T lymphocytes (CTLs) of splenocytes in each group was detected by MTT assay. The levels of tumor necrosis factor-α (TNF-α) and interferon-γ (γ-interferon, γ-INF) expression levels. Results The results of flow cytometry showed that GM-CSF secreting hepatocarcinoma vaccine significantly increased the expression of CD8 + T cells in peripheral blood of mice compared with that of hepatocellular carcinoma (P <0.01). The results of MTT showed that, (P <0.01). The results of Western blot showed that the GM-CSF secreting hepatocarcinoma vaccine significantly increased the cytotoxic activity of splenic cytotoxic T lymphocytes (P <0.01) Down-regulated the expression of TNF-α and γ-INF protein (P <0.01). CONCLUSION: GM-CSF secreting hepatocarcinoma cell vaccine can significantly inhibit the activity of H22 hepatocellular carcinoma cell line H22. The mechanism may be through the activation of CD8 + T cell expression and the cytotoxic activity of splenocyte cytotoxic T lymphocytes, and the decrease of TNF-α And γ-INF protein expression.