IBV M基因与IL-18基因共表达DNA疫苗的免疫原性  

Enhancement of DNA vaccine potency against IBV by co-expression of IBV M and IL-18 gene

作　　者：唐梦君[1] 王红宁[1] 周生[1] 徐永莉[1] 余协中[1] 鲜凌瑾[1] 

机构地区：[1]四川农业大学动物科技学院,四川雅安625014

出　　处：《中国兽医学报》2008年第7期757-761,共5页Chinese Journal of Veterinary Science

基　　金：国家“863”计划资助项目(2003AA2411202005AA246050)

摘　　要：将禽传染性支气管炎病毒(IBV)的M基因和禽白介素18(IL-18)基因分别插入双顺反子质粒pIRES-EGFP/DsRed中,构建IBV M和IL-18基因的共表达质粒pIRES-M/IL18及表达M基因的pIRES-M质粒。通过脂质体转染Vero细胞,利用RT-PCR及间接免疫荧光检测质粒在体外的表达。将构建的质粒用脂质体包裹后,通过腿部肌肉多点注射免疫7日龄雏鸡,28日龄加强免疫1次;免疫前后均采血检测ELISA抗体效价和外周血CD3+、CD4+、CD8+T淋巴细胞的数量;二免后2周用IBV肾型强毒进行攻毒。结果显示,pIRES-M/IL18、pIRES-M质粒均在Vero细胞中有效地转录并表达目的蛋白。从免疫后7 d起,pIRES-M/IL18免疫组产生的抗体水平及外周血T淋巴细胞亚群数量均高于pIRES-M组。pIRES-M/IL18、pIRES-M组对强毒的攻击保护率分别为65%和40%。以上结果表明,禽源IL-18基因与IBV M基因共表达可增强鸡体对DNA疫苗的免疫应答,提高对IBV强毒的抵抗力。The M gene of avian infectious bronchitis virus and interleukin-18(IL-18) gene from chicken were inserted into the bicistronic pIRES-EGFP/DsRed plasmid.The pIRES-M and pIRES-S1/IL18 plasmid encoding M and IL-18 gene were constructed.Plasmids were transfected into the Vero cells by using lipofectamine,respectively,and the expressed products were detected by RT-PCR and indirect immunofluorescence assay.The 7-day-old chickens were immunized intramuscularly with plasmids encapsulated by liposome and boosted thr...

关 键 词：禽传染性支气管炎病毒 M基因 IL-18 DNA疫苗 

分 类 号：S852.65[农业科学—基础兽医学]