Ding Zhuang, Li Zhi-jie, Zhang Xiao-dong, Li Ya-gang, Liu Chang-jun, Zhang Yan-Ping, Li Yang
College of Animal Science and Veterinary Medicine, Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, Jilin University, Changchun 130062, PR China.
Vet Immunol Immunopathol. 2012 Jan 15;145(1-2):206-13. doi: 10.1016/j.vetimm.2011.11.005. Epub 2011 Nov 17.
Viral infections usually result in alterations in the host cell proteome, which determine the fate of infected cells and the progress of pathogenesis. To uncover cellular protein responses in porcine reproductive and respiratory syndrome virus (PRRSV), infected pulmonary alveolar macrophages (PAMs) and Marc-145 cells were subjected to proteomic analysis involving two-dimensional electrophoresis (2-DE) followed by MALDI-TOF-MS/MS identification. Altered expression of 44 protein spots in infected cells was identified in 2D gels, of which the 29 characterised by MALDI-TOF-MS/MS included 17 up-regulated and 12 down-regulated proteins. Some of these proteins were further confirmed at the mRNA level using real-time RT-PCR. Moreover, Western blot analysis confirmed the up-regulation of HSP27, vimentin and the down-regulation of galectin-1. Our study is the first attempt to analyze the cellular protein profile of PRRSV-infected Marc-145 cells using proteomics to provide valuable information about the effects of PRRSV-induced alterations on Marc-145 cell function. Further study of the affected proteins may facilitate our understanding of the mechanisms of PRRSV infection and pathogenesis.
病毒感染通常会导致宿主细胞蛋白质组发生改变，这决定了受感染细胞的命运和发病机制的进展。为了揭示猪繁殖与呼吸综合征病毒（PRRSV）感染时细胞的蛋白质反应，对感染的肺泡巨噬细胞（PAM）和Marc-145细胞进行了蛋白质组学分析，包括二维电泳（2-DE），随后进行基质辅助激光解吸电离飞行时间串联质谱（MALDI-TOF-MS/MS）鉴定。在二维凝胶中鉴定出感染细胞中44个蛋白点的表达发生了改变，其中通过MALDI-TOF-MS/MS鉴定的29个蛋白点包括17个上调蛋白和12个下调蛋白。其中一些蛋白在mRNA水平上通过实时逆转录聚合酶链反应（real-time RT-PCR）进一步得到证实。此外，蛋白质印迹分析证实了热休克蛋白27（HSP27）、波形蛋白的上调以及半乳糖凝集素-1的下调。我们的研究首次尝试利用蛋白质组学分析PRRSV感染的Marc-145细胞的细胞蛋白质谱，以提供有关PRRSV诱导的改变对Marc-145细胞功能影响的有价值信息。对受影响蛋白质的进一步研究可能有助于我们理解PRRSV感染和发病机制。
