鞭毛蛋白及鸡GM-CSF重组新城疫病毒对鸡的免疫增强作用  

Adjuvant effects of flagellin and chicken GM-CSF expressed by recombinant Newcastle disease vaccine

作　　者：王卉[1] 张天援[1] 吕政[1] 孙田[1] 尹杰超[1] 吴云舟[1] 刘云野[1] 吴金 任桂萍[1] 李德山[1] 

机构地区：[1]东北农业大学生命科学学院/生物制药教研室,黑龙江哈尔滨150030 [2]吉林和元生物工程有限公司,吉林吉林132000

出　　处：《中国预防兽医学报》2015年第1期40-43,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省成果产业化项目(1252CGZH29);国家自然基金东北农业大学生物学理科基地科研训练及科研能力提高项目(J1210069/J0124);东北农业大学博士启动基金(2012RCB43)

摘　　要：为评价鼠伤寒沙门氏菌鞭毛蛋白(FliC)以及鸡GM-CSF(chGM-CSF)对新城疫病毒(NDV)弱毒疫苗的免疫增强效果,本实验通过PCR扩增fliC,并合成chGM-CSF编码基因,分别构建重组质粒pBrClone30-fliC和pBrClone30-GM-CSF,拯救获得重组病毒rClone30-fliC和rClone30-GM-CSF。RT-PCR和ELISA检测结果表明:fliC和chGM-CSF基因正确插入重组病毒,并获得稳定表达;重组病毒免疫7 d后重组NDV HI抗体效价检测显示,实验组平均HI效价分别达到4.5 log2和4.8 log2,而r Clone30组和空白组HI效价仅为3 log2和1 log2;免疫7 d后攻毒,空白组SPF鸡均死亡,rClone30组有两例发病,实验组SPF鸡全部存活并且无任何临床症状。本研究结果表明,FliC和chGM-CSF作为佐剂能够有效提高弱毒疫苗的免疫效果。To evaluate the immune enhancement of flagellin （FIiC） and chicken GM-CSF （chGM-CSF） to the attenuated New castle disease （ND） vaccine, the genes of Salmonella typhimurium fliC and chGM-CSF were amplified by PCR and inserted between F and HN NDV vector to construct pBrClone30-fliC and pBrClone30-GM-CSF the recombinant viruses of rClone30-fliC and rClone30-GM-CSF were rescued by reverse-genetics technique. The RT-PCR and ELISA detections indicated that fliC and chGM-CSF genes were stably integrated in the rescued viruses and effectively expressed in cells infected by the recombinant viruses. Additionally, the HI antibody titers against NDV in experimental groups immunized rClone30-fliC and rCIone30-GM-CSF reached 4.51og2 and 4.81og2 at 7 days post vaccination, while rClone30 group and the control group were 31og2 and llog2, respectively. Furthermore, the recombinant viruses provided better protections for chickens against the challenge than that of the parental virus.

关 键 词：鞭毛蛋白 鸡GM-CSF 新城疫病毒 反向遗传操作技术 

分 类 号：S852.65[农业科学—基础兽医学]