pcDNA3.1+/MAGE-3 DNA疫苗诱导特异性抗肿瘤免疫应答的实验研究  

Specific antitumor immune response induced by pcDNA3.1+/MAGE-3 DNA vaccine in mice

作　　者：刘杏娥[1] 孙晓东[2] 吴金民[3] 

机构地区：[1]浙江医院肿瘤科,浙江杭州310016 [2]浙江大学医学院附属邵逸夫医院普外科,浙江杭州310016 [3]浙江大学医学院附属邵逸夫医院肿瘤中心,浙江杭州310016

出　　处：《中国病理生理杂志》2005年第1期167-171,共5页Chinese Journal of Pathophysiology

摘　　要：目的构建pcDNA3.1+/MAGE-3DNA疫苗,观察其在小鼠体内诱导特异性抗肿瘤免疫应答的能力。方法通过RT-PCR构建重组表达质粒pcDNA31+/MAGE-3;以pcDNA31+/MAGE-3DNA疫苗免疫已接种肿瘤细胞的小鼠,每10d重复免疫1次,共3次,以pcDNA31+、PBS为对照。末次免疫后5d检测血清中MAGE-3抗体滴度、小鼠脾淋巴细胞的细胞毒T细胞(cytotoxicTlymphocytes,CTL)杀伤活性、细胞因子IL-2和IFN-γ的浓度,同时计算抑瘤率。结果成功构建了pcDNA31+/MAGE-3DNA疫苗,用此疫苗免疫已接种B16/MAGE-3细胞的小鼠后,能诱导小鼠脾淋巴细胞MAGE-3特异性的杀伤活性,脾细胞培养上清中细胞因子IL-2和IFN-γ的浓度明显增高,血清中抗MAGE-3抗体在1∶20滴度时阳性,肿瘤生长被显著抑制,与pcDNA31+组、PBS组相比,差异显著(P<001)。结论成功构建了pcDNA31+/MAGE-3DNA疫苗,该疫苗在小鼠体内既能激活CTL杀伤活性和CD4+T细胞活性,又能激活体液免疫反应,从而诱导出特异性的抗肿瘤免疫应答。AIM: To construct pcDNA3.1+/MAGE-3 DNA vaccine and investigate the antigen-specific antitumor immune responses induced by pcDNA3.1+/MAGE-3 DNA vaccine in vivo. METHODS: C57BL/6 mice challenged with B16/MAGE-3 cells were immunized by intramuscular injection of pcDNA3.1+/MAGE-3 DNA vaccine every 10 days. pcDNA3.1+ plasmid and PBS were used as controls. After three cycles of immunization, murine splenic lymphocytes, serum, and tumor were obtained for cytotoxity assay, detections of cytokines (IL-2 and IFN-γ), measurement of MAGE-3 antibody, and tumor inhibitory rates, respectively. RESULTS: The pcDNA3.1+/MAGE-3 DNA vaccine immunized murine lymphocytes induced specific cytotoxicity against B16/MAGE-3 cells. Significantly increased secretions of IL-2 and IFN-γ were detected. The titres of antibody against MAGE-3 were 1∶1 and 1∶20, while controls were negative. The tumor inhibitory rate in pcDNA3.1+/MAGE-3 group was significantly different from that in controls. CONCLUSION: The pcDNA3.1+/MAGE-3 DNA vaccine was constructed successfully. pcDNA3.1+/MAGE-3 DNA vaccine activates both cellular and humoral immune responses, and induces antigen-specific antitumor immune responses in vivo. [

关 键 词：黑色素瘤 疫苗 DNA 免疫应答 T淋巴细胞 细胞毒性 

分 类 号：R363[医药卫生—病理学]