PDF(Pubmed)
Abstract：
Serum amyloid A (SAA) proteins belong to a family of acute-phase reactants, playing an integral role in defending the organism from pathological damage. Despite a wealth of data on the regulation of SAA transcripts in teleosts, there is only limited information on these proteins\' abundance in fish. The aim of this study is to characterise SAA protein levels in salmonids using a newly developed antibody specific to salmonid SAA. The salmonid SAA antibody detected SAA and accurately discriminated between stimulated and control specimens from rainbow trout macrophage cell line (RTS-11) in vitro, as well as rainbow trout challenged with Aeromonas salmonicida- or flagellin-stimulated Atlantic salmon in vivo. The presence of SAA protein was analysed in RTS-11 cell line supernatants, liver, and spleen samples using ELISA, immunoblotting, and immunohistochemistry. This study is the first to characterise SAA protein levels in salmonids in vivo and in vitro. The newly developed salmonid SAA antibody was able to discriminate between stimulated and unstimulated specimens, showing that it can be used to study the acute-phase response in salmonids with the potential to be further developed into assays to monitor and evaluate health in wild and farmed fish.
摘要：
血清淀粉样蛋白A（SAA）蛋白属于急性期反应物家族，在保护生物体免受病理性损害方面发挥着不可或缺的作用。尽管有大量关于硬骨鱼SAA成绩单监管的数据，关于鱼类中这些蛋白质丰度的信息有限。这项研究的目的是使用新开发的对鲑鱼SAA具有特异性的抗体来表征鲑鱼中的SAA蛋白水平。鲑鱼SAA抗体检测到SAA，并在体外准确区分了来自虹鳟鱼巨噬细胞系（RTS-11）的刺激标本和对照标本，以及虹鳟鱼在体内受到沙门氏菌气单胞菌或鞭毛蛋白刺激的大西洋鲑鱼的挑战。在RTS-11细胞系上清液中分析SAA蛋白的存在，肝脏,和脾脏样本使用ELISA，免疫印迹,和免疫组织化学。这项研究是第一个在体内和体外表征沙门氏菌中SAA蛋白水平的研究。新开发的salmonidSAA抗体能够区分刺激和未刺激的标本，表明它可用于研究鲑鱼的急性期反应，并有可能进一步发展为监测和评估野生和养殖鱼类健康的分析方法。
