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(CEA)基因和IL-2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IL-2基因片段连接于真核双表达质粒pIRES中,用肌内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情况及其对小鼠脾细胞CEA特异性细胞免疫和体液免疫反应的激活效果。结果:小鼠经肌内注射质粒后,免疫组化证实该核酸疫苗可在体内有效表达CEA分子;免疫检测显示注射后小鼠血清中出现抗CEA抗体,脾细胞IL-4分泌增强,特异性淋巴细胞增值反应明显并伴有自然杀伤细胞NK活性显著增高。结论:实验所构建的核酸疫苗pIRES-CEA,pIRES-IL-2,pIRES-IL-2等可在小鼠体内高效表达并表现出良好的免疫原性。
Purpose： To identify the expression of the DNA vaccine in miee museles and detecl its ability to induce soecuak cekkykar abd gynirak unnybe resoibse to carcinoemhryonic antigens.Methods：Using cloning technique,the cDNA fragments of CEA gene and mouse co-stimulatrory molecule IL-2 gene were constructed into the mammalian co-expression plasmid vector pIRES. Tongue muscle of mice were injected with our plasmid.The expression of CEA molcules in muscle was identified by immunobistochemitry teclnique. The humoral responses were detected by ELISA technique,the special celluar immuse by immunobistochemitry technique.The humoral responses were detected by ELISA technique,the special celluar immuse responses were detected by CTL and NK cytotoxity assay.Results： We identified that our identified that our plasmids can express the CEA and IL-2 molecules in museles of mice.CEA-antibody and IL-4 molecules appear in blood serum of mice,CTL activities and NKcytotoxity were much stronger in immunized mice.Conclusions：The constructed mammalian co-ex-pression plasmid pIRES-CEA, pIRES-IL-2,pIRES-CEA-IL-2 can express CEA and IL-2 molecules in museles and can generate cellular and bumoral immunological responses to CEA positive carcinoma.
出处
《中国癌症杂志》
CAS
CSCD
2005年第4期309-312,共4页
China Oncology
基金
国家自然科学基金资助项目(No:30070744)。
