端粒酶逆转录酶肽-核酸病毒样颗粒疫苗抗肝癌免疫的初步研究  

A preliminary study on anti-liver cancer immunity of the virus-like particulate peptide-nucleic acid vaccine

作　　者：郭红[1] 郝嘉[2] 吴超[3] 房殿春[1] 

机构地区：[1]第三军医大学西南医院消化内科研究所,重庆400038 [2]第三军医大学新桥医院心外科 [3]第三军医大学生物中试基地

出　　处：《中华肝脏病杂志》2006年第4期272-276,共5页Chinese Journal of Hepatology

基　　金：国家自然科学基金(30400208)

摘　　要：目的设计人端粒酶逆转录酶(h T E R T)的新型病毒样颗粒疫苗,并研究其抗肝癌的作用。方法以阳离子抗原肽为桥梁,将人粒细胞、巨噬细胞集落刺激因子和h T E R T克隆入真核双表达载体 p T C A E中,再将多肽和核酸疫苗结合于同一疫苗颗粒,转染并鉴定其免疫原性,评价其转染效率,并观察其免疫转基因小鼠后激发特异性细胞毒性T淋巴细胞(CTL)反应的有效性。结果该疫苗可激发特异性 CTL反应,并对肝癌细胞具有特异性杀伤活性。结论成功构建了具有特异性CTL杀伤活性的hTERT肽核酸病毒样颗粒疫苗,为进一步探索其体内抗肝癌免疫作用奠定了基础。Objectives To construct a novel virus-like particulate peptide-nucleic acid vaccine （VPNV） of human telomerase reverse transcriptase （hTERT）, and to study its anti-liver cancer immunity. Methods A cationic antigenic peptide was synthesized and purified, and then human granulocyte macrophage colony stimulating factor （hGM- CSF） and TERT gene were cloned into the eukaryotic expression vector pTCAE. The peptide was combined with the nucleic acid vaccine to make a VPNV, which was transfected into eukaryotic cell COS-7. The inimunogenicity of hGMCSF and hTERT were detected using ELISA and Western blot. The efficacy of VPNV for inducing antigen specific CTL response was determined using the lactate dehydrogenase release method. Results VPNV was verified capable to trigger specific CTL responses and has shown a specific cytolytic activity to liver cancer cell HepG2. Conclusion A VPNV which can stimulate antigen specific CTL response was successfully constructed. This paves the way for our further investigation of anti-liver cancer immunity in mice.

关 键 词：癌 肝细胞 癌症疫苗 T淋巴细胞 细胞毒性 端粒酶逆转录酶 

分 类 号：R735.7[医药卫生—肿瘤]