摘要
为了探明大鲵虹彩病毒病灭活疫苗的安全性与免疫效力,采用β-丙内酯(β-propiolactone,BPL)灭活鲤上皮瘤细胞(Epithelioma papilloma cyprini,EPC)内增殖的大鲵虹彩病毒(giant salamander iridovirus,GSIV),制备成大鲵虹彩病毒病灭活疫苗,利用免疫剂量为1.0×10^(6) TCID_(50)/mL的灭活疫苗腹腔注射健康大鲵(Andrias davidianus),研究疫苗的安全性和免疫效力。结果显示:一次单剂量注射、单剂量重复注射和超剂量注射后,大鲵的行为和摄食均正常,未见不良反应,证明了制备的大鲵虹彩病毒病灭活疫苗安全;用不同剂量的疫苗(0.1、0.2、0.3、0.4、0.5 mL/尾)免疫健康大鲵,在免疫后第28 d用GSIV进行攻毒,当免疫剂量为0.1 mL/尾时,相对免疫保护率为44.44%,其它4种免疫剂量的相对免疫保护率相似,均高于70%。大鲵在免疫后7 d已开始产生免疫应答,21 d血清抗体效价达到最高值252.48±28.01。在免疫后两周内,MyD88基因、TLR9基因和MHCIIA基因均显著上调。免疫后150 d进行攻毒实验,大鲵的相对免疫保护率为62.5%。二次免疫实验中,大鲵分别在第一次免疫7、14、21、28、56 d后进行第二次免疫。结果显示,第一次免疫21 d后进行二次免疫,其血清抗体效价可以较长时间维持在最高水平,最高血清抗体效价为274.37±25.23,第二次免疫后的第150 d用GSIV进行攻毒,存活率为86.67%。研究结果表明,大鲵虹彩病毒病灭活疫苗最佳免疫方式为腹腔注射疫苗0.2 mL/尾后,21 d进行第二次免疫,免疫剂量为0.2 mL/尾。
The giant salamander iridovirus(GSIV)propagated in the Epithelioma papilloma cyprini(EPC)was inactivated byβ-propiolactone(BPL)to prepare the inactivated vaccines against giant salamander iridovirus disease.To study the safety and immunization efficacy of the vaccine,9-months-old healthy A.davidianus were immunized by inactivated vaccine at a dose of 1.0×10^(6) TCID_(50)/mL.The results showed that A.davidianus with single dose vaccine inoculation,repeated dose vaccine inoculation,and overdose vaccine inoculation showed no abnormalities,which indicated that the developed vaccine was safe for A.davidianus.Vaccines with different doses(0.1,0.2,0.3,0.4,0.5 mL/ind)were used to estimate the best vaccination dose.At 28 days post immunization,challenge experiments were carried to evaluate the efficiency of the vaccine with different doses.When immunized with 0.1 mL/ind,the relative immune protection rate(RPS)was 44.44%,while the other immunized doses were all higher than 70%.At 7 days post vaccination,the serum antibody titers could be detected in immunized A.davidianus,and reached to 252.48±28.01 at 21 days post vaccination.The MyD88 gene,the TLR9 gene and the MHCIIA gene were significantly up-regulated after vaccination.The efficiency of vaccine was tested at the 150th day after immunization.After challenge,the RPS of vaccinated group was 62.5%.In the boost immunization,the A.davidianus were boost immunized at 7,14,21,28 and 56 days post prime immunization.When boost immunization at 21 days post the prime immunization,the serum antibody titers could reach to the levels as high as 274.37±25.23.The challenge experiment was carried out at 150 days post boost immunization.The Survival rate of 21-days boost immunization group was 86.67%.Based on our results,the optimal immunization method for the vaccines of A.davidianus iridovirus disease was intraperitoneal injection of the vaccine 0.2 mL/ind,and boost immunization with the same dose at the 21th day post prime immunization.
作者
索钰杰
林格
江南
李逸群
范玉顶
刘文枝
孟彦
薛明洋
曾令兵
周勇
SUO Yu-jie;LIN Ge;JIANG Nan;LI Yi-qun;FAN Yu-ding;LIU Wen-zhi;MENG Yan;XUE Ming-yang;ZENG Ling-bing;ZHOU Yong(Yangtze River Fisheries Research Institute,Chinese Academy of Fishery Sciences,Wuhan 430223,China;College of Fisheries,Huazhong Agricultural University,Wuhan 430070,China)
出处
《淡水渔业》
CSCD
北大核心
2021年第4期34-41,共8页
Freshwater Fisheries
基金
中国水产科学研究院中央级公益性科研院所基本科研业务费专项资金(2020XT0401、2020TD44)
国家自然科学基金(31702033)。
