Du Cheng, Liu Hai-Fang, Lin Yue-Zhi, Wang Xue-Feng, Ma Jian, Li Yi-Jing, Wang Xiaojun, Zhou Jian-Hua
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, P. R. China.
Department of Preventive Veterinary Medicine, College of Veterinary Medicine, Northeast Agricultural University, Harbin, P. R. China.
Proteomics. 2015 Jun;15(11):1843-58. doi: 10.1002/pmic.201400279. Epub 2015 Mar 21.
Similar to the well-studied viruses human immunodeficiency virus (HIV)-1 and simian immunodeficiency virus (SIV), equine infectious anemia virus (EIAV) is another member of the Lentivirus genus in the family Retroviridae. Previous studies revealed that interactions between EIAV and the host resulted in viral evolution in pathogenicity and immunogenicity, as well as adaptation to the host. Proteomic analysis has been performed to examine changes in protein expression and/or modification in host cells infected with viruses and has revealed useful information for virus-host interactions. In this study, altered protein expression in equine monocyte-derived macrophages (eMDMs, the principle target cell of EIAV in vivo) infected with the EIAV pathogenic strain EIAV(DLV34) (DLV34) was examined using 2D-LC-MS/MS coupled with the iTRAQ labeling technique. The expression levels of 210 cellular proteins were identified to be significantly upregulated or downregulated by infection with DLV34. Alterations in protein expression were confirmed by examining the mRNA levels of eight selected proteins using quantitative real-time reverse-transcription PCR, and by verifying the levels of ten selected proteins using parallel reaction monitoring (PRM). Further analysis of GO and Kyoto Encyclopedia of Genes and Genomes (KEGG)-Pathway enrichment demonstrated that these differentially expressed proteins are primarily related to the biological processes of oxidative phosphorylation, protein folding, RNA splicing, and ubiquitylation. Our results can facilitate a better understanding of the host response to EIAV infection and the cellular processes required for EIAV replication and pathogenesis.
与经过充分研究的人类免疫缺陷病毒1型（HIV-1）和猴免疫缺陷病毒（SIV）类似，马传染性贫血病毒（EIAV）是逆转录病毒科慢病毒属的另一个成员。先前的研究表明，EIAV与宿主之间的相互作用导致了病毒在致病性和免疫原性方面的进化，以及对宿主的适应性。蛋白质组学分析已被用于检测感染病毒的宿主细胞中蛋白质表达和/或修饰的变化，并揭示了病毒与宿主相互作用的有用信息。在本研究中，使用二维液相色谱-串联质谱（2D-LC-MS/MS）结合同位素标记相对和绝对定量（iTRAQ）技术，检测了感染EIAV致病株EIAV(DLV34)（DLV34）的马单核细胞衍生巨噬细胞（eMDM，EIAV在体内的主要靶细胞）中蛋白质表达的变化。经鉴定，210种细胞蛋白的表达水平因感染DLV34而显著上调或下调。通过定量实时逆转录PCR检测8种选定蛋白的mRNA水平，并通过平行反应监测（PRM）验证10种选定蛋白的水平，证实了蛋白质表达的变化。对基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集的进一步分析表明，这些差异表达的蛋白质主要与氧化磷酸化、蛋白质折叠、RNA剪接和泛素化的生物学过程相关。我们的结果有助于更好地理解宿主对EIAV感染的反应以及EIAV复制和发病机制所需的细胞过程。
