重组干扰素、趋化因子合剂对草鱼免疫保护作用  

Lmmunoprotection of grass carp(Ctenopharyngodon idella) with a mixture of recombinant IFN and CCL4 against GCHV infection

作　　者：吴初新[1] 李东明[2,3] 余新建[2] 黄胜和[2,3] 胡成钰[2] 

机构地区：[1]南昌师范高等专科学校,江西南昌330103 [2]南昌大学生命科学与食品工程学院,江西南昌330031 [3]南昌大学抚州医学分院,江西抚州344000

出　　处：《南昌大学学报（理科版）》2014年第1期91-95,共5页Journal of Nanchang University(Natural Science)

基　　金：江西省科技计划项目(No.20111BBF60020);江西省自然科学基金项目(20132BAB204021)

摘　　要：草鱼细菌性和病毒性疾病严重制约着草鱼养殖生产的发展。干扰素和趋化因子为鱼类重要的非特异性免疫因子,参与鱼类抗病毒、免疫调节等反应。在已克隆的1个草鱼Ⅰ型干扰素基因(GU139255)和CCL4基因(EU918199)的基础上,通过大肠杆菌表达系统分别获得了其相应的表达蛋白。利用喷雾法将这两种蛋白分别或混合添加到饲料中并投喂草鱼,利用攻毒实验验证这两种蛋白对草鱼的免疫保护作用。结果显示:单独添加重组IFN或CCL4后能有效提高草鱼存活率,免疫21 d时其成活率分别为63.33%和46.67%。当同时添加重组IFN和CCL4后,免疫保护效果则更为明显,21 d时草鱼存活率为73.33%,免疫保护率达69.23%。而且,重组IFN和CCL4能够诱导内源IFN、IRF1、STAT3、CCL4表达上调,而两者混合的蛋白诱导效果更为强烈。提示混合的Ⅰ型干扰素和CCL4重组蛋白可以作为一种新型鱼类免疫制剂。Bacterial or viral diseases had seriously damaged grass carp breeding industry in China. Interferon and chemokines were the important non-specific immune-related cytokines,which involved in disease resistance response in fish. Previously,typeⅠIFN gene（ GU139255） and CCL4 gene（ EU918199） had been cloned in our laboratory. Here,we expressed IFN or CCL4 protein by E. coli expression systems. Then,these purified recombinant proteins were sprayed and mixed into the feed. The results showed that recombinant IFN（ rCiIFN） or CCL4（ rCiCCL4） alone could effectively improve the survival of grass carp,which reached 63. 33% and 46. 67% at 21d,respectively. After stimulation with rCiIFN ＋ rCiCCL4,the immune protective effect was more obvious. The survival of grass carp was 73. 33%,and relative percent survival value reached 69. 23% at 21d. RT-PCR analysis also showed that IFN, IRF1,STAT3,and CCL4 gene were significantly up-regulated by rCiIFN ＋ rCiCCL4. It indicated that the mixture of rCiIFN and rCiCCL4 could be used as a new type of fish immune drugs.

关 键 词：IFN CCL4 免疫保护 免疫制剂 草鱼 

分 类 号：Q786[生物学—分子生物学]