Development of an inactivated iridovirus vaccine against turbot viral reddish body syndrome

Turbot(Scophthalmus maximus L.) reddish body iridovirus(TRBIV) was propagated in turbot fin cells(TF cells) and inactivated as the TRBIV vaccine with its protection efficiency evaluated in this study.TF cells were cultured in 10% bovine calf serum(BCS)-containing MEM medium(pH7.0) at 22℃,in which TRBIV propagated to a titer as high as 105.6 TCID50 mL-1.The TRBIV was inactivated with 0.1% formalin and formulated with 0.5% aluminum hydroxide.The inactivated vaccine caused neither cytopathogenic effect(CPE) on TF cells nor pathogenic effect on turbots.After being administered with the vaccine twice via muscle injection,the turbot developed high-tittered TRBIV neutralizing antibodies in a dose-dependent manner.The vaccine protected the turbot from dying with an immunoprotection rate of 83.3% as was determined via subcutaneous vaccination in the laboratory and 90.5% via bath vaccination in turbot farms,respectively.The inactivated vaccine was very immunogenic,efficiently preventing tur-bot from death.It holds the potential of being applied in aquaculture.     

鱼类口服疫苗的研究进展

口服免疫是通过胃肠粘膜系统进行抗原呈递.较传统的注射和浸浴免疫简易便捷.是更为理想的免疫途径。一般认为鱼的后肠是摄取抗原的主要部位,口服疫苗引起的免疫应答以粘膜免疫为主.系统免疫应答并不是很强。在口服疫苗的研制过程中,出现了各种问题.使得这种疫苗迄今无法问世。本文围绕鱼类口服免疫的意义.作用机理以及口服疫苗的研究现状作一综述.     

Vaccination in three different ways against vibriosis of <Emphasis Type="Italic">Seriola dumerili</Emphasis> caused by <Emphasis Type="Italic">Vibrio hollisae</Emphasis>

Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili （amberjack） suffering from vibriosis. Healthy S. dumeriIi were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1：40 at 7 d, and reached its peak of 1：320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of 〈1：10, 1：20, 1：160 were observed at 7 d, 14 d, 35 d respectively. Compared with the control, the vaccinated fish exhibited significantly higher lysozyme activities （P〈0.05）. Upon challenge with virulent strain, the relative percent survival （RPS） of injected, immersed and oral administrated fish were 75%, 45%, and 40% respectively, and the injected fish showed significantly higher RPS than immersed and oral administrated fish. The results suggested that vaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm.     

三联灭活细菌疫苗诱导的大黄鱼肾组织SMART cDNA文库的构建

大黄鱼(Pseudos ciaenacrocea(Richardson))是我国特有的经济鱼种,以其味道鲜美,营养丰富和极高的药用价值而受到青睐,在大黄鱼人工育苗和网箱养殖等技术相继成熟之后,人工养殖得到迅猛发展,仅福建省2000年就有养殖网箱约30万[1].由于养殖规模的不断扩大和养殖环境日益恶化,导致大面积细菌性疾病的爆发[2~5],给大黄鱼的养殖业造成了巨大的损失.对于细菌性疾病的防治,目前主要使用抗生素.     

利用酒精酵母细胞表面展示技术制备活疫苗及其潜在应用

最近几年,出现把有关的基因在宿主表达之后的目的蛋白分泌并固定在细菌、病毒和酵母菌细胞表面上的技术一表面展示技术。在这些宿主中,酒精酵母菌是最佳的微生物。本文对利用酒精酵母细胞表面展示技术制备活疫苗的基本原理,在酵母细胞表面展示蛋白质的优点,以及哈维氏弧菌溶血素蛋白在酵母菌细胞表面上展示及其作为活疫苗的潜在应用作了综述。研究结果表明在酒精酵母菌细胞表面上展示的哈维氏弧菌溶血素蛋白在海洋动物中具有很好的免疫原性和免疫保护性。     

猪传染性胃肠炎病毒S、N基因核酸疫苗的构建与免疫原性试验

采用RT—PCR扩增TGEVSCH株S基因5’端主要抗原位点编码区和全长N基因,将其插入真核表达载体pVAXl,构建单独与嵌合表达S、N基因的pVAX—S、pVAX—N、pVAX—S—N3种重组质粒。通过脂质体将重组质粒体外转染COS-7细胞,利用间接免疫荧光检测目的基因的表达情况。将6周龄NIH小鼠随机分为5组,分别于腿部肌肉注射重组质粒pVAX、pVAX—S、pVAXS-N、pVAX—S＋pVAX—N与pVAX—S＋pVAX,共免疫3次,间隔2周。通过间接EI。IsA以及流式细胞仪分别检测免疫小鼠血清中特异性TGEVIgG抗体和外周血T淋巴细胞亚群数量。结果显示,重组质粒构建正确且在COS-7细胞中得以表达;pVAX—S＋pVAX—N质粒混合免疫组与pVAX—s＋pVAx质粒混合免疫对照组相比,可有效提高免疫小鼠外周血T淋巴细胞亚群数量和血清中特异性抗体水平的增加,而pVAX—S-N免疫组与pVAX—S组比较,未能起到提高免疫小鼠血清中特异性抗体水平的和外周血T淋巴细胞亚群数量作用。以上结果表明S、N基因的联合应用可有效加强TGEV核酸疫苗的免疫效力,但这种增强作用与S、N基因联合应用的方式直接相关。     

溶藻弧菌疫苗对凡纳滨对虾免疫功能的影响

应用溶藻弧菌疫苗饲喂凡纳滨对虾,测定了弧菌疫苗对凡纳滨对虾免疫指标的影响以及 疫苗的免疫保护作用。结果发现,投喂弧菌疫苗后,溶菌活力在12～20d内迅速增强,在20d时 活力达到最高,VF1组高于VF2和VF3组,但VF2和VF3之间差异不显著;凝集效价在投喂后的 第8天开始升高,在14d时达到最高,实验组的凝集活力均高于对照组,但实验组之间无明显的差 异;而超氧化物歧化酶(SOD)活力和溶血活力变化不明显。以每克饲料含疫苗细胞1.0×109、 1.0×108和1.0×107个的浓度投喂,对凡纳滨对虾的免疫保护率分别为66.7%、46.7%和13.3%。 可见弧菌疫苗对凡纳滨对虾的免疫保护率随疫苗浓度的增大而升高。     

Protective immunity of orange-spotted grouper(Epinephelus coioids) against a nervous necrosis virus isolated from China,and determination of the complete sequences of the virus

1 IntroductionNervous necrosis virus( NNV) is one of theworldwide pathogens in cultured species and breed-ing from the late 1980s (Bovo et al.,1999; Tan etal., 2001; Breuil et al., 2001; Munday et al.,2002; Lin et al., 2001). The virus is isometric andnon…     

禽流感病毒抗原表位分析及H5N1亚型基因工程疫苗设计

以亚洲地区H5N1亚型禽流感病毒(Avian Influenze Virus)流行株为研究对象,利用计算机软件,对同源性较高的HA1(Hemagglutnin,HA,血凝素)和NP(Nucleocapsid protein,核衣壳蛋白)进行全基因序列分析,优选出HA1蛋白的主要T细胞表位和B细胞表位,以及NP蛋白的主要CTL(Cytotoxicity T lymphocyte,细胞毒性T淋巴细胞)表位.依据这些优选表位,设计了禽流感病毒H5N1亚型基因工程疫苗.构建了基因工程疫苗表达载体pRSET-AIV,外源基因能够在大肠杆菌表达系统中得到良好表达.表达产物免疫小鼠后,血清中IgA和IgG抗体水平明显上升,在体外培养脾细胞可产生IL-2、IL-4和IFN-γ细胞因子.验证了该H5N1亚型基因工程疫苗的抗原性,证实该基因工程疫苗在免疫小鼠体内激发体液免疫的同时调动了细胞免疫.     

Vaccination in three different ways against vibriosis of Seriola dumerili caused by Vibrio hollisae

Bacterin was prepared by formalin-inactivating the virulent strain of Vibrio hollisae isolated from diseased Seriola dumerili (amberjack) suffering from vibriosis. Healthy S. dumerili were vaccinated by respective procedures of intramuscular injection, immersion, and orally administration. Results of the three different vaccinations were compared. Blood was drawn from the vaccinated fish every 7 days, and the antibody titers and lysozyme activities of the sera were determined. The antibody titer of injected fish was 1:40 at 7 d, and reached its peak of 1:320 at 28 d, while the fish vaccinated by immersion and orally administration exhibited weak antibody responses, the antibody titres of <1:10, 1:20, 1:160 were observed at 7 d, 14 d, 35 d respectively. Compared with the control, the vaccinated fish exhibited significantly higher lysozyme activities (P<0.05). Upon challenge with virulent strain, the relative percent survival (RPS) of injected, immersed and oral administrated fish were 75%, 45%, and 40% respectively, and the injected fish showed significantly higher RPS than immersed and oral administrated fish. The results suggested that vaccination of S. dumerili by the injection would be the best strategy to prevent the vibriosis in S. dumerili farm.