Key Laboratory for Veterinary Bio-Product Engineering, Ministry of Agriculture and Rural Affairs, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
School of Food and Biological Engineering, Jiangsu University, Zhenjiang 212013, China.
Viruses. 2021 Jul 2;13(7):1293. doi: 10.3390/v13071293.
Caprine herpesvirus 1 (CpHV-1) is a member of the alpha subfamily of herpesviruses, which is responsible for genital lesions and latent infections in goat populations worldwide. In this study, for the first time, the transcriptome and proteomics of CpHV-1 infected Madin Darby bovine kidney (MDBK) cells were explored using RNA-Sequencing (RNA-Seq) and isobaric tags for relative and absolute quantitation-liquid chromatography tandem mass spectrometry (iTRAQ-LC-MS/MS) technology, respectively. RNA-Seq analysis revealed 81 up-regulated and 19 down-regulated differentially expressed genes (DEGs) between infected and mock-infected MDBK cells. Bioinformatics analysis revealed that most of these DEGs were mainly involved in the innate immune response, especially the interferon stimulated genes (ISGs). Gene Ontology (GO) enrichment analysis results indicated that the identified DEGs were significantly mainly enriched for response to virus, defense response to virus, response to biotic stimulus and regulation of innate immune response. Viral carcinogenesis, the RIG-I-like receptor signaling pathway, the cytosolic DNA-sensing pathway and pathways associated with several viral infections were found to be significantly enriched in the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database. Eleven selected DEGs (Mx1, RSAD2, IFIT1, IFIT2, IFIT5, IFIH1, IFITM3, IRF7, IRF9, OAS1X and OAS1Y) associated with immune responses were selected, and they exhibited a concordant direction both in RNA-Seq and quantitative real-time RT-PCR analysis. Proteomic analysis also showed significant up-regulation of innate immunity-related proteins. GO analysis showed that the differentially expressed proteins were mostly enriched in defense response and response to virus, and the pathways associated with viral infection were enriched under KEGG analysis. Protein-protein interaction network analysis indicated most of the DEGs related to innate immune responses, as DDX58(RIG-I), IFIH1(MDA5), IRF7, Mx1, RSAD2, OAS1 and IFIT1, were located in the core of the network and highly connected with other DGEs. Our findings support the notion that CpHV-1 infection induced the transcription and protein expression alterations of a series of genes related to host innate immune response, which helps to elucidate the resistance of host cells to viral infection and to clarify the pathogenesis of CpHV-1.
山羊疱疹病毒 1(CpHV-1)是疱疹病毒α亚科的成员,在全球范围内导致山羊生殖道病变和潜伏感染。在这项研究中,我们首次分别使用 RNA 测序(RNA-Seq)和同位素标记相对和绝对定量-液相色谱串联质谱(iTRAQ-LC-MS/MS)技术,研究了 CpHV-1 感染 Madin Darby 牛肾(MDBK)细胞的转录组和蛋白质组学。RNA-Seq 分析显示,感染和模拟感染的 MDBK 细胞之间有 81 个上调和 19 个下调的差异表达基因(DEGs)。生物信息学分析表明,这些 DEGs 主要参与固有免疫反应,特别是干扰素刺激基因(ISGs)。基因本体论(GO)富集分析结果表明,鉴定出的 DEGs 主要富集于对病毒的反应、对病毒的防御反应、对生物刺激的反应和对固有免疫反应的调节。病毒致癌作用、RIG-I 样受体信号通路、胞质 DNA 感应途径以及与几种病毒感染相关的途径在京都基因与基因组百科全书(KEGG)通路数据库中显著富集。还发现了与免疫反应相关的 11 个选定的 DEGs(Mx1、RSAD2、IFIT1、IFIT2、IFIT5、IFIH1、IFITM3、IRF7、IRF9、OAS1X 和 OAS1Y),它们在 RNA-Seq 和定量实时 RT-PCR 分析中均显示出一致的方向。蛋白质组学分析还显示固有免疫相关蛋白的显著上调。GO 分析表明,差异表达蛋白主要富集于防御反应和对病毒的反应,KEGG 分析中富集了与病毒感染相关的途径。蛋白质-蛋白质相互作用网络分析表明,大多数与固有免疫反应相关的 DEGs,如 DDX58(RIG-I)、IFIH1(MDA5)、IRF7、Mx1、RSAD2、OAS1 和 IFIT1,位于网络的核心部位,并与其他 DGEs 高度连接。我们的研究结果支持以下观点,即 CpHV-1 感染诱导了一系列与宿主固有免疫反应相关的基因的转录和蛋白表达改变,这有助于阐明宿主细胞对病毒感染的抵抗力,并阐明 CpHV-1 的发病机制。