pcDNA3.1/MAGE-3-HSP70融合蛋白诱导特异性抗肿瘤免疫应答的研究  

Specific antitumor immune response induced by pcDNA3.1/MAGE-3-HSP70 fusion DNA vaccine

作　　者：刘杏娥[1] 孙晓东[2] 吴金民[2] 

机构地区：[1]浙江医院肿瘤科,杭州310013 [2]浙江大学医学院附属邵逸夫医院普外科,杭州310016

出　　处：《中华微生物学和免疫学杂志》2005年第12期998-1001,共4页Chinese Journal of Microbiology and Immunology

摘　　要：目的构建重组表达质粒pcDNA3.1/MAGE-3-HSP70,观察其诱导免疫应答的能力和抗肿瘤免疫治疗的作用。方法通过RT-PCR扩增MAGE-3和HSP70cDNA,以pcDNA3.1为载体,构建pcDNA3.1/MAGE-3-HSP70重组质粒,肌肉注射接种小鼠,间隔7d,共3次,以pcDNA3.1/MAGE-3、pcD-NA3.1/HSP70、pcDNA3.1和PBS为对照,检测脾淋巴细胞CTL杀伤活性、脾细胞培养上清IL-2、IFN-γ浓度、外周血淋巴细胞亚群变化及鼠血清中抗MAGE-3抗体水平。pcDNA3.1/MAGE-3-HSP70重组质粒免疫已负荷B16/MAGE-3的小鼠,观察小鼠成瘤时间、生存时间。结果pcDNA3.1/MAGE-3-HSP70质粒免疫的小鼠,其脾淋巴细胞对MAGE-3阳性靶细胞表现出明显的杀伤活性,与各对照组相比,P<0.01,差异有统计学意义;外周血中CD4+、CD8+T细胞和脾细胞培养上清中TH1类细胞因子IFN-γ、IL-2水平明显升高。pcDNA3.1/MAGE-3-HSP70重组质粒免疫已负荷B16/MAGE-3的小鼠后,小鼠成瘤时间明显延迟,生存期明显延长。结论pcDNA3.1/MAGE-3-HSP70DNA疫苗在体内能诱导出显著的MAGE-3特异性抗肿瘤免疫应答,且能抑制体内已经存在的少量肿瘤细胞的成瘤。Objective To construct pcDNA3.1/MAGE-3-HSP70 eukaryotie expression plasmids and to investigate the capability of pcDNA3.1/MAGE-3-HSP70 DNA vaccine to induce specific antitumor immune responses. Methods The HSP70 cDNA, which was amplified by RT-PCR, and the eukaryotie expression plamnid pcDNA3.1/MAGE-3-HSP70 was constructed. C57BL/6 mice were vaccinated with pcDNA3.1/MAGE-3-HSP70 on day 0, 7 and 14, as pcDNA3.1/MAGE-3, pcDNA3.1/HSP70, pcDNA3.1 and PBS for control. Splenocytes CTLs, the levels of eytokines IL-2, IFN-γ and the level of antibodies against MAGE-3 and the changes of the T lymphocyte subsets were checked. The challenged mice were immunized with pcDNA3. 1/MAGE-3-HSP70 plasmid and monitored every two days for the evidence of tumor growth. Results The CTLs activity from the mice immunized with pcDNA3.1/MAGE-3-HSP70 plasmid was significantly different to that of control groups（ P 〈 0.01 ）. The counting of CD4^＋ , CD8^＋ and the levels of IFN-γ, IL-2 significantly increased after immunization with pcD- NA3.1/MAGE-3-HSP70 plasmid as compared with those of control groups（ P 〈 0.01）. After immunization with pcDNA3.1/MAGE-3-HSP70 DNA vaccine, the tumor volume was significantly smaller and the survival time was longer than those of controls. Conclusion The pcDNA3.1/MAGE-3-HSP70 DNA vaccine is able to induce significant specific cellular immune responses with the outcome of eradicating MAGE-3-expressing tumors.

关 键 词：DNA疫苗 MAGE-3 HSP70 肿瘤 免疫应答 

分 类 号：R730.3[医药卫生—肿瘤]