癌胚抗原与γ-干扰素核酸疫苗对机体特异性细胞免疫的激活作用  

CEA nucleic acid vaccine induces special cellular immune response to a carcinoembryonic antigen in mice

作　　者：王中川[1] 傅传刚[1] 何天霖[1] 郝立强[1] 王庆敏[2] 孙树汉[2] 

机构地区：[1]第二军医大学附属长海医院肛肠外科,上海200433 [2]第二军医大学医学遗传学教研室,上海200433

出　　处：《肿瘤防治杂志》2005年第14期1041-1044,共4页China Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金面上项目(30070744)

摘　　要：目的检测以双表达质粒pIRES为载体构建的带有全序列癌胚抗原(carcinoembry-onic antigens,CEA)基因和γ-干扰素(interfer-on-γ,IFN-γ)基因的核苷酸疫苗对机体特异性抗肿瘤免疫反应的激活效果。方法利用分子生物学技术,将CEA基因片段和IFN-γ基因片段连接于真核双表达质粒pIRES中,用肌肉注射方法接种核酸疫苗;检测疫苗在小鼠肌肉组织中的表达及其对小鼠脾细胞CEA特异性细胞免疫反应的激活效果。结果小鼠经肌肉注射质粒后,核酸疫苗可在体内有效表达CEA和IFN-γ分子;小鼠特异性淋巴细胞增殖反应明显增高,并且伴有自然杀伤细胞(natural killercell,NK细胞)活性显著增高,F=15289·67,P<0·001。结论实验所构建的核酸疫苗pIRES-CEA、pIRES-CEA-IFN-γ和pIRES-CEA-IFN-γ等可在小鼠体内高效表达,并表现出良好的细胞免疫原性。OBJECTIVE:To identify the expression of the nucleic acid vaccine in mice muscles and detect its ability to induce special cellular immune response to a carcinoembryonic antigen. METHEDS: By using cloning technique, the cDNA fragments of CEA gene and IFN-γ gene were constructed into the mammalian co-expression plasmid vector pIRES; the reconstructed nucleic acid was injected into mices’s tongne muscle. The expression of CEA molecules in muscle were identified by the immunohistochemitry technique, and the special celluar immuse responses were detected by CTL and NK cytotoxity assay. RESULTS: The plasmid expressed the CEA molecules in mice’s tongue muscles detected by the immunohistochemitry technique. CTL activities and NK cytotoxity were strong in immunized mice, F=15 289.67,P<0.001. CONCLUSION: The constructed mammalian co-expression plasmid pIRES-CEA, pIRES-IFN-γ and pIRES-CEA-IFN-γ can express CEA and IFN-γ molecules in muscles and can generate cellular immunological responses against CEA positive carcinoma.

关 键 词：癌胚抗原/免疫学 干扰素Ⅱ型/免疫学 杀伤细胞 天然 基因表达 聚合酶链反应 

分 类 号：R73-36[医药卫生—肿瘤]