hTERT隐性表位肽-核酸病毒样颗粒疫苗抗胃癌免疫的初步研究  

The anti-gastric cancer immunity of the virus-like particulate peptide-nucleic acid vaccine of human telomerase reverse transcriptase

作　　者：郭红[1] 郝嘉[2] 吴超[3] 石云[3] 房殿春[1] 

机构地区：[1]第三军医大学西南医院全军消化内科中心,重庆400038 [2]第三军医大学新桥医院心血管外科,重庆400037 [3]第三军医大学医学检验系临床微生物及免疫学教研室,重庆400038

出　　处：《第三军医大学学报》2006年第2期169-171,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目(30400208)~~

摘　　要：目的设计新型的人端粒酶逆转录酶(hTERT)肽-核酸病毒样颗粒疫苗,并研究其抗胃癌的作用。方法将阳离子隐性表位肽和DNA疫苗协同性结合于同一疫苗形成病毒样颗粒,通过DNA阻滞试验、DNA沉淀试验、免疫组化等试验优化该疫苗参数,并鉴定其免疫原性和观察其体内激发特异性CTL反应的有效性。结果确定制备疫苗的参数为浓度为150mmol/LNaCl,电荷比为4。证实该疫苗可激发特异性CTL反应,并对胃癌细胞具有特异性杀伤活性,Elispot实验显示能够有效诱导的IFN-γ分泌。结论成功构建了具有特异性CTL杀伤活性的hTERT隐性表位肽-核酸病毒样颗粒疫苗,为进一步探索其体内抗胃癌免疫作用奠定了基础。Objective To construct a novel virus-like particulate peptide-nucleic acid vaccine （VPNV） of human telomerase reverse transcriptase （ hTERT）, and to study the anti-gastric cancer immunity. Methods Cationic antigenic peptide was combined with the DNA vaccine to obtain VPNV. The preparing parameters were optimized by DNA blockade assay, DNA precipitation assay, immunohistochemistry etc. The immunogenicity of hGMCSF and hTERT were detected by ELISA and immunohistochemistry. The CTL response and the IFN-γ, secretion were detected by lactate dehydrogenase release method and enzyme-linked immunospot （ELISPOT） assay. Resuits The parameters of VPNV should be 150 mmoL/L NaCl and at charge ratio of 4. The vaccine was confirmed to trigger effective and specific CTL responses and have specific cytolytic activity to gastric cancer cell KATO-3 and induce IFN-γ, secretion in vivo. Conclusion VPNV that could stimulate antigen specific CTL response was successfully constructed. This paves the way for further investigation of the anti-gastric cancer immunity.

关 键 词：人端粒酶逆转录酶 胃癌 隐性表位 肽-核酸病毒样颗粒疫苗 细胞毒T淋巴细胞 

分 类 号：R392.33[医药卫生—免疫学] R392.7[医药卫生—基础医学]