维氏气单胞菌灭活疫苗对草鱼免疫相关基因表达的影响及其保护效果  

Expression of immunity-associated genes and protective effect in grass carps immunized by inactivated Aeromonas veronii vaccine

作　　者：任燕 时云朵 曾伟伟 王英英 李莹莹 常藕琴 尹纪元 石存斌 REN Yan;SHI Yun-duo;ZENG Wei-wei;WANG Ying-ying;LI Ying-ying;CHANG Ou-qin;YIN Ji-yuan;SHI Cun-bin(Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province,Key Laboratory of Fishery Drug Development of Ministry of Agriculture and Rural Affairs,Pearl River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Guangzhou 510380,Guangdong Province,China)

机构地区：[1]中国水产科学研究院珠江水产研究所广东省水产动物免疫技术重点实验室农业农村部渔用药物创制重点实验室,广东广州510380 [2]四川省水产学校,四川成都611730

出　　处：《中国生物制品学杂志》2019年第7期726-731,共6页Chinese Journal of Biologicals

基　　金：现代农业产业技术体系建设专项资金(CARS-45)

摘　　要：目的探讨维氏气单胞菌(Aeromonas veronii,Av)灭活疫苗对草鱼免疫相关基因表达的影响及保护效果。方法采用GZ09007菌株制备Av灭活疫苗,分别通过浸泡和注射两种途径免疫草鱼,同时设对照组(未免疫),于免疫后2、4、7、11、14和21d,经实时荧光定量PCR法检测草鱼脾脏、头肾、鳃组织中IgM、IL-1β、C3、C-凝集素及溶菌酶的m RNA表达量;于免疫30 d后用同源菌株进行攻毒,连续观察14d,计算各免疫组的相对保护率(relative percent survival,RPS)。结果 2个免疫组的5个免疫相关基因mRNA表达量均显著高于对照组(P <0. 001),且注射组表达量普遍高于浸泡组,注射组IgM基因较浸泡组更早表达,C3、C-凝集素及溶菌酶基因表达量更高或表达持续时间更长。脾脏中IL-1β和溶菌酶基因表达量达最高值为免疫后14~21d,头肾和鳃组织中5个基因表达量达最高值均为免疫后2~4d,各基因表达量最高值为对照组的1.81~37. 52倍。腹腔注射攻毒14 d后,注射组及浸泡组的RPS分别为66.7%和36.7%。结论Av灭活疫苗免疫草鱼后,可增强脾脏、头肾和鳃组织中IgM、IL-1β、C3、C-凝集素及溶菌酶基因mRNA表达,提高草鱼抗Av感染的能力。Objective To investigate the influence of inactivated Aeromonas veronii (Av) vaccine on expression of immunity-associated genes in grass carps as well as protective effect of the vaccine. Methods Inactivated Av vaccine was prepared with GZ09007 strain,with which grass carps were immunized by immersion and intraperitoneal injection respectively,using those unimmunized as control. The expression levels of IgM,IL-1β,C3,C-lectin and lysozyme mRNAs in spleen,head kidney and gill tissues of grass carps 2,4,7,11,14 and 21 d after immunization were determined by real-time fluorescent quantitative PCR. The grass carps were challenged i. p. with a homologous A. veronii strain 30 d after immunization and observed for 14 d to calculate the relative percent survival (RPS). Results The expression levels of five immunity-associated genes in grass carps immunized by two routes were significantly higher than those in control group (P < 0. 001),while were higher in those immunized by injection than in those by immersion. However,IgM gene was expressed earlier in grass carps immunized by injection than in those by immersion,while the expressions of C3,C-lectin and lysozyme genes were at higher levels or more persistent. Except those of IL-1β and lysozyme genes in spleen reached the maximums 14 ~ 21 d,the expression levels of five genes in head kidney and gill tissues reached the maximums 2 ~ 4 d after immunization,and the highest expression levels of various genes were 1. 81 ~ 37. 52 times of those in control group. The RPSs of grass carps immunized by injection and immersion 14 d after challen ge were 66. 7% and 36. 7% respectively Conclusion Inactivated Av vaccine enhanced the expressions of IgM,IL-1β, C3,C-lectin and lysozyme in spleen,head kidney and gill tissues of grass carps as well as the specific immune responses against Av infection.

关 键 词：维氏气单胞菌 灭活疫苗 实时荧光定量PCR 免疫相关基因 免疫保护 

分 类 号：S942.5[农业科学—水产养殖]