摘要
为获知约氏黄杆菌(Flavobacterium johnsoniae)减毒活疫苗(M170株)浸泡免疫鳜(Siniperca chuatsi)后的免疫应答,采用荧光定量RT-PCR方法测定免疫后鳜肝脏溶菌酶(Lys)基因、脾脏免疫球蛋白(Ig M)基因表达动力学,并检测了血清中Lys活性和抗体效价变化规律。结果表明,约氏黄杆菌减毒活疫苗浸泡免疫后,鳜肝脏Lys基因表达量显著上调(P<0.05),第48小时表达量达到峰值,是对照组的10.9倍(P<0.05),直到第28天表达量仍显著高于对照组(P<0.05)。同时免疫鱼血清Lys活性明显升高,第2天达到峰值,是对照组的4.35倍(P<0.05),第6天与对照组水平相当;免疫鳜脾脏Ig M基因相对表达量从第7天开始上调并显著高于对照组(P<0.05),至第28天其相对表达量为对照组的98.7倍(P<0.05);而免疫鱼血清抗体效价在第7天开始高于对照组,第28天达到峰值(26)。免疫后第28天采用强毒株M168进行攻毒,结果显示,免疫组相对成活率显著高于对照组,其相对免疫保护率(relative percentage survival,RPS)达85.9%。结果表明约氏黄杆菌减毒株M170可引起鳜特异性和非特异性免疫应答,并提供良好的免疫保护效果。
To study the immune response of mandarin fish （Siniperca chuatsi） to attenuated Flavobacteriumjohnsoniae strain M170, we used fluorescence quantitative RT-PCR method for determination of liver lysozyme （Lys） gene in spleen, gene expression dynamics of immunoglobulin （IgM） , and detected the changes of the Lys activity and the antibody titer in serum. The results show that after the F. johnsoniae attenuated accine immunization, the Lys gene expression in liver increased significantly （ P 〈 0. 05 ）, reaching the peak at the 48thhour, 10.9 times higher than that of the control group （P 〈 0.05 ）, and had been higher than the control group significantly until the 28th day （P 〈0. 05）. Meanwhile, the Lys activity in serum increased significantly, reaching the peak on the 2nd day, 4. 35 times higher than that of the control group （ P 〈 0. 05 ） and was equivalent to the level of the control group on the 6th day. The relative expression of IgM gene from mandarin fish spleen began to increase and had been significantly higher than that of the control group since the 7th day （P 〈 0. 05 ）, 98.7 times higher than that of the control group until the 28＇h day （P 〈 0. 05 ）. The antibody titer in the immune fish serum was higher than that of the control group significantly since the 7＇h day （P 〈 0. 05 ）, reaching the peak （26） on the 28th day. On the 28th day after immunization, the strain M168 was used to attack the virus, and the relative survival rate of the immu- nized group was higher than that of the control group significantly, and the relative percentage smwival （RPS） was 85.9%. Tile re- suits show that the specific and non-specific immune response can be induced by attenued F. johnsoniae M170 and provide good protection for mandarin fish.
作者
付小哲
方伟
林强
刘礼辉
梁红茹
黄志斌
徐德海
李宁求
FU Xiaozhe FANG Wei LIN Qiang LIU Lihui LIANG Hongru HUANG Zhibin XU Dehai LI Ningqiu(Key Lab. of Fishery Drug Development, Ministry of Agriculture,Key Lab. of Aquatic Animal Immune Technology, Guangdong Province,Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China Guangdong Provincial Aquatic Animal Epidemic Disease Prevention and Control Center, Guangzhou 510220, China Aquatic Animal Health Research Unit, USDA, Auburn AL36832, USA)
出处
《南方水产科学》
CAS
CSCD
北大核心
2017年第5期33-38,共6页
South China Fisheries Science
基金
中国水产科学研究院基本科研业务费专项(2016GH09)
国家科技支撑计划项目(2012BAD25B02)
关键词
鳜
约氏黄杆菌
减毒活疫苗
溶菌酶
免疫球蛋白
相对免疫保护率
Siniperca chuatsi
Flavobacterium johnsoniae
live vaccine
lysozyme
IgM
relative percentage survival （RPS）
