Shivappa R B, McAllister P E, Edwards G H, Santi N, Evensen O, Vakharia V N
Center for Biosystems Research, University of Maryland Biotechnology Institute, College Park, MD 20742, USA.
Dev Biol (Basel). 2005;121:165-74.
Various attempts to develop a vaccine against infectious pancreatic necrosis virus (IPNV) have not yielded consistent results. Thus, at present, no commercial vaccine is available that can be used with confidence to immunize fry of salmon and trout. We generated a cDNA clone of the large genome segment A of an IPNV Sp strain and expressed all structural protein genes in insect cells and larvae using a baculovirus expression system. Green fluorescent protein was also coexpressed as a reporter molecule. High yields of IPNV proteins were obtained and the structural proteins self assembled to form virus-like particles (VLPs). We tested the immunogenicity of the putative VLP antigen in immersion vaccine experiments (two concentrations) in rainbow trout (Oncorhynchus mykiss) fry, and by intraperitoneal immunisation of Atlantic salmon (Salmo salar) pre-smolts using an oil adjuvant formulation. Rainbow trout were challenged by immersion using either the Sp or the VR-299 strain of IPNV two or three weeks post-vaccination, while Atlantic salmon were bath challenged with Sp strain after two months, after parr-smolt transformation. In the rainbow trout fry challenged two weeks post-immunization, cumulative mortality rates three weeks post challenge were 14 % in the fry that had received the highest dose versus 8 % in the control groups. No indication of protection was seen in repeated trials using a lower dose of antigen and challenge three weeks post-immunisation. The cumulative mortality rate of intraperitoneally immunised Atlantic salmon post-smolts four weeks post challenge was lower (56%) than in the control fish (77%), showing a dose-response pattern.
开发针对传染性胰腺坏死病毒(IPNV)疫苗的各种尝试均未取得一致结果。因此,目前尚无可以放心用于免疫鲑鱼和鳟鱼苗的商业疫苗。我们构建了IPNV Sp株大基因组片段A的cDNA克隆,并使用杆状病毒表达系统在昆虫细胞和幼虫中表达了所有结构蛋白基因。绿色荧光蛋白也作为报告分子共表达。获得了高产的IPNV蛋白,并且结构蛋白自组装形成病毒样颗粒(VLP)。我们在虹鳟(Oncorhynchus mykiss)鱼苗的浸泡疫苗实验(两种浓度)中,以及通过使用油佐剂配方对大西洋鲑(Salmo salar)幼鲑进行腹腔免疫,测试了推定的VLP抗原的免疫原性。在接种疫苗后两周或三周,用IPNV的Sp株或VR - 299株通过浸泡对虹鳟进行攻毒,而在仔鱼 - 幼鲑转化两个月后,用Sp株对大西洋鲑进行浸浴攻毒。在免疫后两周攻毒的虹鳟鱼苗中,接受最高剂量疫苗的鱼苗在攻毒后三周的累积死亡率为14%,而对照组为8%。在使用较低剂量抗原并在免疫后三周攻毒的重复试验中未观察到保护迹象。腹腔免疫的大西洋鲑幼鲑在攻毒后四周的累积死亡率(56%)低于对照鱼(77%),呈现出剂量反应模式。