Kania Per W, Chettri Jiwan K, Buchmann Kurt
Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, University of Copenhagen, Stigbojlen 7, DK-1870 Frederiksberg, Denmark.
Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, University of Copenhagen, Stigbojlen 7, DK-1870 Frederiksberg, Denmark.
Fish Shellfish Immunol. 2014 Oct;40(2):648-58. doi: 10.1016/j.fsi.2014.08.018. Epub 2014 Aug 19.
Serum amyloid A (SAA) is an integral part of the innate immune response in mammals and considered to be important during the acute phase response. The present study was undertaken to elucidate the role of SAA protein in the innate immune response of rainbow trout. A monoclonal antibody raised against a recombinant peptide of rainbow trout SAA was characterized using Western blot, dot blot, ELISA and immunohistochemistry. SAA association with high density lipoprotein (HDL) complicated band identification in Western blot, but delipidization of the SAA-HDL isolate highly increased the quality of reaction in the western blot. Rainbow trout fry (87 days post hatch) infected with Yersinia ruckeri showed a significant up-regulation of the SAA gene at 72 h post infection with an increase until 96 h post infection. Non-significant up-regulations were seen at earlier time points i.e. 4 and 24 h. The expression pattern of SAA significantly correlated to the immunohistochemical analysis of the infected fry. A weak staining was seen in liver tissue at 4 h post infection which increased in intensity during the course of infection i.e. 24, 72 and 96 h post infection.
血清淀粉样蛋白A（SAA）是哺乳动物固有免疫反应的一个组成部分，被认为在急性期反应中很重要。本研究旨在阐明SAA蛋白在虹鳟鱼固有免疫反应中的作用。使用蛋白质印迹法、斑点印迹法、酶联免疫吸附测定法和免疫组织化学方法对一种针对虹鳟鱼SAA重组肽产生的单克隆抗体进行了表征。在蛋白质印迹法中，SAA与高密度脂蛋白（HDL）的结合使条带鉴定变得复杂，但SAA-HDL分离物的脱脂处理显著提高了蛋白质印迹法中的反应质量。感染鲁氏耶尔森菌的虹鳟鱼苗（孵化后87天）在感染后72小时显示SAA基因显著上调，并持续增加直至感染后96小时。在较早的时间点即4小时和24小时观察到非显著上调。SAA的表达模式与感染鱼苗的免疫组织化学分析显著相关。感染后4小时在肝脏组织中可见微弱染色，在感染过程中即感染后24小时、72小时和96小时染色强度增加。
