Liu Peng-Fei, Du Yishuai, Meng Lingjie, Li Xian, Liu Ying
Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China; University of Chinese Academy of Sciences, Beijing, 100039, China.
Institute of Oceanology, Chinese Academy of Sciences, Qingdao, 266071, China.
Dev Comp Immunol. 2017 Jul;72:140-153. doi: 10.1016/j.dci.2017.02.013. Epub 2017 Feb 21.
Aeromonas salmonicida is a major etiologic agent which induces furunculosis and is globally harmful in salmonid and turbot cultures, especially in Atlantic salmon (Salmo salar) farming. In order to improve knowledge of its poorly understood pathogenesis, we utilized high-throughput proteomics to display differentially expressed proteins in the kidney of Atlantic salmon challenged with high and low infection dose of A. salmonicida at 7 and 14 days. In quantitative proteomic assays, isobaric tags for relative and absolute quantitation (iTRAQ) combined with 2D LC-MS/MS is emerging as a powerful methodology in the search for disease-specific targets and biomarkers. In this study, 4009 distinct proteins (unused ≥ 1.3, which is a confidence ≥ 95%) were identified in three two-dimensional LC/MS/MS analyses. Then we chose 140 proteins (fold change ratio ≥ 1.5 and P < 0.01) combined with protein-protein interaction analysis to ultimately obtain 39 proteins in network which could be considered as potential biomarkers for Atlantic salmon immune responses. Nine significant differentially expressed proteins were consistent with those at the proteomic level used to validate genes at the transcriptomic level by qPCR. Collectively, these data was first reported using an iTRAQ approach to provide additional elements for consideration in the pathophysiology of A. salmonicida and pave the way to resolve the influence of this disease in Atlantic salmon.
杀鲑气单胞菌是引发疖疮病的主要病原体，在全球范围内对鲑科鱼类和大菱鲆养殖有害，尤其是在大西洋鲑（Salmo salar）养殖中。为了增进对其了解甚少的发病机制的认识，我们利用高通量蛋白质组学技术，展示了在7天和14天时受到高感染剂量和低感染剂量杀鲑气单胞菌攻击的大西洋鲑肾脏中差异表达的蛋白质。在定量蛋白质组学分析中，用于相对和绝对定量的等压标签（iTRAQ）与二维液相色谱-串联质谱联用，正成为寻找疾病特异性靶点和生物标志物的强大方法。在本研究中，通过三次二维液相色谱/串联质谱分析鉴定出4009种不同的蛋白质（未使用值≥1.3，置信度≥95%）。然后，我们选择了140种蛋白质（变化倍数≥1.5且P<0.01），结合蛋白质-蛋白质相互作用分析，最终在网络中获得39种可被视为大西洋鲑免疫反应潜在生物标志物的蛋白质。9种显著差异表达的蛋白质与蛋白质组学水平上用于通过qPCR在转录组水平验证基因的蛋白质一致。总体而言，这些数据首次使用iTRAQ方法报道，为杀鲑气单胞菌病理生理学的研究提供了更多需要考虑的因素，并为解决这种疾病对大西洋鲑的影响铺平了道路。
