目的 对新型肺癌基因疫苗的免疫原性进行评价。 方法 利用Furin 2A将黑色素瘤抗原A3(MAGE-A3)基因与GM-CSF基因进行连接,克隆入真核表达载体pVAX1,构建基因疫苗pVAX1- MAGE-A3-F2A-GM-CSF。将该疫苗体外转染293T细胞,利用免疫印迹和ELISA的方法对其表达情况进行检测。然后,将疫苗及对照组分别免疫接种小鼠,通过ELISA和ELISPOT法初步检测其诱导的体液免疫和细胞免疫反应。 结果 成功克隆并构建了肺癌基因疫苗pVAX1- MAGE-A3-F2A-GM-CSF,并证实了其在真核细胞中表达。疫苗免疫接种小鼠模型后的检测结果显示,与空载体对照组以及单独抗原组相比,pVAX1- MAGE-A3-F2A-GM-CSF能够诱导产生更强的体液免疫反应和细胞免疫反应。结论 pVAX1- MAGE-A3-F2A-GM-CSF能够同时诱导较强的体液免疫和细胞免疫反应,其抗肿瘤效应将通过进一步实验深入研究。
Abstract
Objective To construct a recombinant lung cancer DNA vaccine with high expression antigen MAGE-A3 in lung cancer and to evaluate its immunogenicity.Methods MAGE-A3 genes were fused with GM-CSF by F2A and cloned into pVAX1 to construct the gene vaccine ppVAX1-MAGE-A3-F2A-GM-CSF. After the gene vaccine was transfected into 293T cells, the expression of the gene vaccine was detected by Western blot and ELISA. Then, the gene vaccine and the control group were injected into the mouse model, and the immune activity was detected by ELISA and ELISPOT.Results Gene vaccine pVAX1-MAGE-A3-F2A-GM-CSF was constructed, and the expression of the antigen was confirmed. After the mice were vaccinated with the gene vaccine, the test Results of ELISA and ELISPOT showed that pVAX1-MAGE-A3-F2A-GM-CSF induced both humoral immune response and cellular immune response compared with the control group.Conclusions pVAX1-MAGE-A3-F2A-GM-CSF can induce both humoral immune response and cellular immune response, and its potential anti-tumor effect needs to be further studied.
关键词
黑色素瘤相关抗原 A3 (MAGE-A3) /
肺癌 /
粒细胞-巨噬细胞集落刺激因子 /
基因疫苗
Key words
MAGE-A3 /
lung cancer /
granulocyte-macrophage colony-stimulating factor /
gene vaccine
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