Orpetveit Irene, Gjøen Tor, Sindre Hilde, Dannevig Birgit H
National Veterinary Institute, P.O. Box 8156, Dep, 0033, Oslo, Norway.
Arch Virol. 2008;153(3):485-93. doi: 10.1007/s00705-007-0018-1. Epub 2008 Jan 3.
The early interactions between infectious pancreatic necrosis virus (IPNV) from Atlantic salmon and susceptible cell lines were studied using a virus overlay protein binding assay (VOPBA). Membrane preparations from four different cell lines, bluegill fry (BF)-2 cells, Chinook salmon embryo (CHSE)-214 cells, salmon head kidney (SHK)-1 cells and Atlantic salmon kidney (ASK) cells were separated by SDS-PAGE, blotted to nitrocellulose membranes and incubated with either a highly virulent IPNV field isolate or different recombinant IPNV strains exhibiting variations in virulence. Binding of virus to the respective membrane fractions was detected using either polyclonal or monoclonal antibodies. Independent of variations in virulence, IPNV bound in a specific manner to an approximately 220-kDa membrane protein from the salmonid cell lines CHSE-214, SHK-1 and ASK, whereas the size of the binding protein from the non-salmonid cell line BF-2 was approximately 190 kDa. Neutralization of IPNV prior to incubation with the blotted membrane fractions inhibited binding to the indicated proteins. Complete deglycosylation of the membrane proteins did not interfere with the binding of IPNV, suggesting that the interaction between virus and cells is not carbohydrate specific. The described binding proteins may represent putative cellular binding sites or receptors for IPNV.
利用病毒覆盖蛋白结合试验（VOPBA）研究了来自大西洋鲑鱼的传染性胰腺坏死病毒（IPNV）与易感细胞系之间的早期相互作用。将来自四种不同细胞系的膜制剂，即蓝鳃太阳鱼（BF）-2细胞、奇努克鲑鱼胚胎（CHSE）-214细胞、鲑鱼头肾（SHK）-1细胞和大西洋鲑鱼肾（ASK）细胞，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）进行分离，转移至硝酸纤维素膜上，然后与高毒力的IPNV野外分离株或不同毒力的重组IPNV毒株一起孵育。使用多克隆抗体或单克隆抗体检测病毒与各个膜组分的结合。与毒力变化无关，IPNV以特异性方式结合到鲑鱼细胞系CHSE-214、SHK-1和ASK中一种约220 kDa的膜蛋白上，而非鲑鱼细胞系BF-2中结合蛋白的大小约为190 kDa。在与印迹膜组分孵育之前对IPNV进行中和可抑制其与指定蛋白质的结合。膜蛋白的完全去糖基化并不干扰IPNV的结合，这表明病毒与细胞之间的相互作用不是碳水化合物特异性的。所述的结合蛋白可能代表IPNV假定的细胞结合位点或受体。
