摘要
目的:观察核酸疫苗pIRES-CEA-HSP70诱导小鼠产生的细胞免疫应答,以探索结肠癌术后复发的防治新途径。方法:将pIRES、pIRES-CEA、pIRES-HSP70、pIRES-CEA-HSP70制作核酸疫苗免疫小鼠,采用MTT比色法,检测小鼠脾淋巴细胞对结肠癌细胞CW-2的杀伤效应。结果:核酸疫苗pIRES-CEA-HSP70免疫组的小鼠脾淋巴细胞,在效靶比50∶1、25∶1、12.5∶1、6.25∶1下,对CW-2的杀伤率分别为(35.43±3.88)%(、26.88±3.17)%(、20.33±1.92)%(、15.55±0.76)%,均明显高于其余3组,差异有统计学显著性(P<0.01)。pIRES、pIRES-CEA、pIRES-HSP70 3组杀伤率差异无显著性(P>0.05)。结论:核酸疫苗pIRES-CEA-HSP70可诱导小鼠产生特异性细胞免疫应答,为其用于结肠癌的治疗提供实验基础。
Objective：To investigate cellular immune response in mice induced by pIRES-CEA-HSP70 fusion nucleic acid vaccine and explore a new way to prevent postoperative relapse of colon cancer. Methods：Plasmids pIRES, pIRES-CEA, pIRES- HSP70,and pIRES-CEA-HSP70 were transfected into CHO cells by Lipofectamine 2000. The transfectants were selected by G418. The expression of CEA and HSP70 were detected at transcription and translation levels by reverse transcriptase polymerase chain reaction （RT-PCR） and enzyme linked immunosorbent assay （ELISA）. Mice were immunized with the four vaccines. The cytotoxic effect of the spleen lymphocytes isolated from immunized mice on CW 2 colon cancer cells was measured by MTT assay. Results：RT-PCR and ELISA assay demonstrated positive expression of CEA and HSP70 in CHO cells. The killing rate of CW-2 cells by the spleen lymphocytes in pIRES-CEA-HSP70-immunized mice was （35.43±3.88） %, （26.88±3.17）%, （20.33±1.92）%, and （15.55±0.76）% at effecto-target ratio of 50 ： 1,25 ： 1,12.5 ： 1, and 6.25 ： 1, respectively, which was remarkably higher than the other three groups immunized with pIRES, pIRES-CEA, and pIRES-HSP70 nucleic acid vaccines, respectively （P〈0.01）. The killing rate between the other three groups was not significantly different（P〉0.05）. Conclusion： The pIRES-CEA-HSP70 nucleic acid vaccine can induce specific cellular immune response in mice, which establishes the experimental basis for the treatment of colon cancer.
出处
《肿瘤》
CAS
CSCD
北大核心
2006年第3期241-243,共3页
Tumor
基金
福建省自然科学基金资助项目(编号:C0410043)
关键词
结肠肿瘤
癌胚抗原
热休克蛋白质类70
疫苗
DNA
细胞毒性
免疫
Colonic neoplasms
Carcinoembryonic antigen
Heat shock proteins 70
Vaccine,DNA
Cytotoxicity,immunologic
