Hou Libo, Xiu Yunji, Wang Jian, Liu Xiaoqian, Liu Yuhan, Gu Wei, Wang Wen, Meng Qingguo
Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210023, China; Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
Jiangsu Key Laboratory for Biodiversity & Biotechnology, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210023, China; Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, Shandong, China; Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences, Nanjing Normal University, 1 Wenyuan Road, Nanjing 210046, China.
J Proteomics. 2016 Mar 16;136:112-22. doi: 10.1016/j.jprot.2015.12.026. Epub 2015 Dec 31.
Spiroplasma eriocheiris, as a novel aquaculture pathogen, has led into catastrophic economic losses in aquaculture. The Macrobrachium rosenbergii hemocytes were major target cells in S. eriocheiris infection. Our study was designed to examine the hemocytes' immune response at the protein levels. The differential proteomes of the prawn hemocytes were analyzed immediately prior to injection with the pathogen, and at 192h post-injection by isobaric tags for relative and absolute quantization (iTRAQ) labeling, followed by liquid chromatogramphytandem mass spectrometry (LC-MS/MS). A total of 69 differentially expressed proteins were identified. Forty-nine proteins were up-regulated and 20 proteins were down-regulated resulting from a S. eriocheiris infection. Up-regulated proteins included vertebrate gliacolin-like protein, vitellogenin, Gram-negative binding protein 1, alpha2 macroglobulin isoform 2 (a2M), etc. Down-regulated proteins, involved with beta-1,3-glucan-binding protein (BGBP), immunoglobulin like, Rab7, lipopolysaccharide and β-1,3-glucan (LGBP), actin-related protein, etc. Selected bioactive factors (tachylectin, α2M and vitellogenin, BGBP, C-type lectin, LGBP and Rab7) were verified by their immune roles in the S. eriocheiris infection using real-time PCR. The present work could serve as a basis for future studies on the proteins implicated in the susceptibility/resistance of M. rosenbergii to S. eriocheiris, as well as contribute to our understanding of disease processes in prawns.
This is the first time using an iTRAQ approach to analyze proteomes of M. rosenbergii mobilized against S. eriocheiris infection and substantiated the hemocytes' proteomic changes in M. rosenbergii using an infection model. The results reported here can provide a significant step forward toward a more complete elucidation of the immune relationship between M. rosenbergii and the pathogen S. eriocheiris.
罗氏沼虾支原体作为一种新型水产养殖病原体，已给水产养殖业带来了灾难性的经济损失。罗氏沼虾血细胞是罗氏沼虾支原体感染的主要靶细胞。我们的研究旨在从蛋白质水平检测血细胞的免疫反应。在注射病原体之前以及注射后192小时，通过相对和绝对定量的等压标签（iTRAQ）标记，随后进行液相色谱串联质谱（LC-MS/MS）分析对虾血细胞的差异蛋白质组。共鉴定出69种差异表达蛋白。罗氏沼虾支原体感染导致49种蛋白上调，20种蛋白下调。上调的蛋白包括脊椎动物胶质蛋白样蛋白、卵黄蛋白原、革兰氏阴性结合蛋白1、α2巨球蛋白亚型2（a2M）等。下调的蛋白涉及β-1,3-葡聚糖结合蛋白（BGBP）、免疫球蛋白样、Rab7、脂多糖和β-1,3-葡聚糖（LGBP）、肌动蛋白相关蛋白等。通过实时PCR验证了所选生物活性因子（速激肽、α2M和卵黄蛋白原、BGBP、C型凝集素、LGBP和Rab7）在罗氏沼虾支原体感染中的免疫作用。目前的工作可为今后研究罗氏沼虾对罗氏沼虾支原体易感性/抗性相关蛋白奠定基础，也有助于我们了解对虾的疾病过程。
这是首次使用iTRAQ方法分析罗氏沼虾针对罗氏沼虾支原体感染的蛋白质组，并通过感染模型证实了罗氏沼虾血细胞的蛋白质组变化。本文报道的结果可为更全面阐明罗氏沼虾与病原体罗氏沼虾支原体之间的免疫关系迈出重要一步。
