黑色素瘤抗原-1诱导特异性抗肝癌免疫应答

Cancer testis antigen MAGE-1 can induce immune response for anti-tumor in vivo and in vitro

目的:探讨黑色素瘤抗原-1(MAGE-1)对特异性抗肝癌免疫应答反应的影响。方法:分别用转染pcD-NA3.1-MAGE-1(重组子)、pcDNA3.1(空质粒)以及未行转染的人肝癌细胞SMMC-7721冻融抗原致敏树突状细胞(DC),诱导T淋巴细胞增殖分化为细胞毒性T淋巴细胞(CTL)。乳酸脱氢酶释放法检测CTL对SMMC-7721细胞的杀伤活性。48只C57BL小鼠随机分为重组子组、空质粒组和PBS组,每组16只。用pcDNA3.1-MAGE-1预先免疫重组子组小鼠,1次/10d,第3次免疫后第5天ELISA法检测各组8只小鼠脾细胞培养液中IFN-γ和IL-2的含量,第6天各组8只小鼠接种鼠源性肝癌细胞H22,观察其生存时间。结果:未转染、空质粒和重组子组诱导的CTL对靶细胞的杀伤率以及各组小鼠股四头肌MAGE-1mRNA表达的比较差异有统计学意义(F=139.601和538.650,P﹤0.001),重组子组高于未转染和空质粒组。PBS组和空质粒组小鼠脾细胞培养液中IFN-γ和IL-2的含量为0,重组子组小鼠脾细胞培养液中IFN-γ含量为(372.33±10.08)ng/L,IL-2含量为(108.67±12.81)ng/L。3组小鼠荷瘤生存时间比较差异有统计学意义(F=31.837,P<0.001),重组子组小鼠生存时间明显延长。结论:MAGE-1可诱导特异性抗肿瘤免疫应答,从而显著延长荷瘤小鼠的生存时间。

Aim： To observe the immune anti-tumor function of MAGE-1 in vivo and in vitro. Melhods：Cytotoxic T lymphocytes（CTLs） were activated by DCs pulsed with freeze-thaw antigen of SMMC-7721-MAGE-1 or SMMC-7721-pcD- NA3.1 （ - ） or wild type SMMC-7721, respectively. Taking wild type SMMC-7721 as target cells, the cytotoxicity of CTLs was detected by use of cytoTo non-radioactive cytotoxicity assay, pcDNA3. 1-MAGE-1 was injected intramuscularly into C57BL mice on the 1st, llth and 21st day with 100 t^g/mouse, taking pcDNA3.1 and PBS as controls. On the 26th day, the volume of IL-2 and IFN-r/which were secreted by splenocytes was detected by ELISA. On the 27th day, the mice were inoculated mouse hepatoma H22 ceils by celiac injection. The survival time of each mouse was recorded. Results： The ex- pression of MAGE-1 mRNA and the cytotoxicity of CTLs activated by DCs pulsed with freeze-thaw antigen of SMMC-7721- MAGE-1 were higher than those of SMMC-7721-pcDNA3. 1 （ - ） group and SMMC-7721 group （F = 139. 601 and 538.650,P 〈0.001）. The contents of IL-2 and IFN-~, in pcDNA3.1-MAGE-1 group were （ 108.67 ± 12.81 ） ng/L and （372.33 ± 10.08 ） ng/L, respectively, while the contents of control groups were 0 ng/L. The survival time of pcDNA3.1- MAGE-1 group was （ 17.4 ＋ 1.6） d, longer than those of the pcDNA3.1 group [ （ 13.1± 1.7） days] and the PBS group [ （ 12.0 ± 1.1 ） days] （ F = 31. 837, P 〈 0. 001 ）. Conclusion ： The pcDNA3.1-MAGE-1 DNA vaccine is able to induce cellular immune responses in vivo and in vitro.