【作者】 任燕； 时云朵； 曾伟伟； 王庆； 王英英； 李莹莹； 常藕琴； 尹纪元； 石存斌；

【Author】 REN Yan;SHI Yunduo;ZENG Weiwei;WANG qing;WANG Yingying;LI Yingying;CHANG Ouqin;YIN Jiyuan;SHI Cunbin;Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Key Laboratory of Fishery Drug Development of Ministry of Agriculture,Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences;

【机构】 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/农业部渔用药物创制重点实验室； 四川省水产学校；

【摘要】 为研究维氏气单胞菌灭活疫苗免疫效果,分别通过浸泡和注射两种途径免疫草鱼,通过实时荧光定量PCR检测免疫后2 d、4 d、7 d、11 d、14 d和21 d脾脏、头肾、鳃中IgM、IL-1β、C3、C-凝集素和溶菌酶的mRNA表达量,在免疫30d后用同源菌株攻毒试验鱼来测定各免疫组的相对保护率。结果显示,免疫组的5个免疫相关基因的相对表达量显著高于对照组(P<0.05),注射组Ig M基因较浸泡组更早表达,补体C3、C-凝集素以及溶菌酶的基因表达量更高或表达持续时间更长。免疫后各组织中的5个基因显著上调表达,且注射组表达量普遍高于浸泡组;除了脾脏中IL-1β和溶菌酶基因表达量最大值在免疫后14d-21d,头肾和鳃组织中5个基因表达量最大值均在免疫后2d-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后显示,注射组的相对保护率达到66.7%,浸泡组的达到36.7%。结果表明,维氏气单胞菌灭活疫苗浸泡或注射免疫草鱼,增强了脾脏、头肾和鳃组织中Ig M、IL-1β、C3、C-凝集素和溶菌酶基因表达,提高了草鱼抗维氏气单胞菌感染的能力。更多还原

【Abstract】 Formalin-inactivated Aeromonas veroniivaccine was administered to grass carps through immersion and intraperitoneal injection, respectively. Spleen, head kidney and gill tissues were sampled on days 2, 4, 7, 11, 14 and 21 vaccination, and the expression quantities of IgM, IL-1β, C3, C-lectin and lysozyme m RNAs in the tissues were detected through the real-time fluorescence quantitative PCR. Formalin-inactivated A. veroniivaccine was administered just once, then, the fish were challenged intraperitoneally with lethal A. veroniion 30^th daypost-vaccination. The result of immersion and intraperitoneal injectionvaccination showed thatimmune grass carp exhibited enhanced expression of genes that are involved in both innate and adaptive immune responsescompared to saline control. The results showed that, the expression of all the tested genes varied significantly. In the three organs, the expression ofIgM, IL-1β, C3, C-lectin and lysozyme genes reached the peak between 2 d-4 d but IL-1β and lysozyme genes reached the peak between 14 d-21 d in spleen. The gene maximum expression was 1.81-37.5 times higher than that of control group.The relative percent survival（RPS） of intraperitoneal injectionimmunization was found to be 66.7% on day 30 post-vaccination, while immersion immunization with the vaccine followed by an experimental challenge gave an RPS of 36.7%. These results indicate that Formalin-inactivated A. veroniivaccine administered to grass carps can significantly promote the gene expression of IgM, IL-1β, C3, C-lectin and lysozyme in spleen, head kidney and gill tissues. It can enhance specific immune responses against A. veronii infection.更多还原