Clade 2.3.2e H5亚型禽流感DNA疫苗的免疫保护效力研究  

Protective efficacy of DNA vaccine against clade 2.3.2e H5N1 avian influenza virus

作　　者：潘俊慧 梁真洁 陈普成[1] 唐猛[1] 曾显营[1] 柳金雄[1] 邓国华[1] 姜永萍[1] 陈化兰[1] PAN Jun-hui;LIANG Zhen-jie;CHEN Pu-cheng;TANG Meng;ZENG Xian-ying;LIU Jin-xiong;DENG Guo-hua;JIANG Yong-ping;CHEN Hua-lan(National Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,CAAS,Harbin 150069,China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨黑龙江150069

出　　处：《中国预防兽医学报》2018年第10期930-933,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金："十二五"农村领域国家科技计划(2015BAD12B03/001);哈市科技攻关项目(2013AA6BN001)

摘　　要：为研制和更新针对H5亚型禽流感病毒(AIV)流行株的DNA疫苗,本研究将新近分离的H5亚型AIV Clade2.3.2e分支代表株A/Duck/Anhui/S1246/2015 (DK/AH/S1246/15)密码子优化的HA基因定向克隆至载体pCAGGS中构建重组质粒pCA-S1246,将该质粒转染293T细胞。利用间接免疫荧光和western blot检测,结果显示, HA蛋白可以在293T细胞中正确表达。将15μg、 30μg和60μg的p CA-S1246质粒分别免疫3周龄的SPF鸡, 3周以后以相同的剂量加强免疫后1周,检测其HI抗体平均效价分别可达1∶56、 1∶16和1∶37;加强免疫1周后用105EID50的DK/AH/S1246/15进行攻击时,免疫组的保护率为100%。本研究为DNA质粒pCA-S1246作为防控Clade.2.3.2e AIV的候选DNA疫苗株提供了实验依据。The constant mutation of hemagglutinin （HA） gene of highly pathogenic avian influenza virus （HPAIV）, resulting in antigenic variation, compromises the vaccine immune effects, and there is a pressing need to reconstruct and update candidate vaccine strain according to the HPAIV epidemiological results. In this study, the codon optimized HA gene of latest isolated A/Duck/Anhui/S1246/2015 （DK/AH/S1246/15） （H5N1）, a representative strain of clade 2.3.2e HPAIVs, was subcloned into pCAGGS vector to construct the recombinant plasmid pCA-S1246 as the DNA vaccine against AIV. After being transfected into 293T cells, the HA expressed effectively detected by IFA and western blot. In addition, groups of SPF chickens were immunized with the doses of 15μg, 30μg and 60μg of pCA-S1246, respectively, and boosted with same dose after three weeks to evaluate the protective efficacy of the candidate DNA vaccine. The blood samples were collected from the immunized chickens in each dose group for the antibody detection, and the average HI titers were 1：56 of 15μg, 1∶16 of 30μg and 1∶37 of 60μg, respectively, 1 week after boost. Our results showed that pCA-S1246 was able to protect chicken from being infected by DK/AH/S1246/15 completely, and the pCA-S1246 has the potential to be a vaccine candidate against clade 2.3.2e H5N1 virus.

关 键 词：DNA疫苗 Clade.2.3.2e禽流感病毒 免疫效力评价 

分 类 号：S852.65[农业科学—基础兽医学]