鸡胚成纤维细胞体外筛选CpG寡聚脱氧核苷酸免疫活性分子的研究  

Screening of chicken CpG oligonucleotide using chicken embryo fibroblast in vitro

作　　者：刘伟杰[1,2] 张琳[1] 吴家强[1] 孔正杰 张秀美[1] 黄艳艳[1] 许传田[1] 杨少华[1] 黄庆华[1] 常维山[2] 

机构地区：[1]山东省农业科学院畜牧兽医研究所山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [2]山东农业大学动物科技学院,山东泰安271018

出　　处：《中国兽医科学》2016年第5期649-655,共7页Chinese Veterinary Science

基　　金："十二五"农村领域国家科技计划项目(2015BAD12B03)

摘　　要：为了优化鸡特异性CpG ODN活性分子体外筛选体系,避免核酸细胞毒性的干扰,增强细胞应答效果,尝试利用CEF代替鸡淋巴细胞进行CpG ODN活性分子的体外筛选。将设计的9条不同序列的CpG ODN分别刺激CEF,通过测定细胞增殖效果和相关免疫基因的表达变化,评判免疫应答效果是否准确反映了CpG ODN结构特征,确定CpG ODN最佳免疫刺激活性分子。结果显示,CEF对CpG ODN的刺激呈现出良好的免疫反应,正确体现了CpG ODN序列的构效关系,成功筛选到3条CpG ODN活性分子,表明CEF适合于CpG ODN免疫佐剂的体外筛选。To avoid the interference of the cytotoxicity of nucleic acid and enhance the effects of immune re- sponse, CEF was used to screen chicken CpG ODNs in vitro instead of chicken lymphocytes.CEF were stimulated by nine designed CpG ODNs, respectively,then cell proliferation and fold changes in related immune gene expression were assayed,which was compared with the reaction of lymphocytes.As a result, strong immune responses were ex- hibited on CEF stimulated by CpG ODNs ,which was correlated with the structure of CpG ODNs.Finally,three CpG ODNs were selected for in yiyo test.Present study demonstrated that CEF was suitable for the screening of CpG ODNs in vitro.

关 键 词：鸡胚成纤维细胞 CPG ODN 体外筛选 细胞毒性 免疫应答 

分 类 号：S852.43[农业科学—基础兽医学]