Transcriptome analysis reveals heat shock protein 70 (HSP70) induced by tembusu virus infection is associated with immune responses.

D Zhao, K Han, L Zhang, X Huang, Q Liu, J Yang, Y Liu, Y Li, F Wu
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Abstract

The outbreak and prevalence of tembusu virus (TMUV) endanger the breeding industry of waterfowls. However, little is known about the molecular mechanism underlying TMUV infection. It was reported that heat shock protein 70 (HSP70) was a positive regulator of the infection of TMUV. In order to study the interactions between HSP70 and host immune response to TMUV infection, TMUV-infected cells with or without HSP70 inhibitor were harvested and subjected to deep sequencing to identify genes differentially expressed. We found 43 differentially expressed genes (DEGs) in HSP70 inhibitor-treated and mock-treated TMUV-infected DF-1 cells. Of these DEGs, 39 genes were down-regulated significantly. Gene Ontology analysis suggested that the DEGs were mainly involved in biological process, cellular component and molecular function. Kyoto Encyclopedia of Genes and Genomes analysis showed that the DEGs mainly related to the activation of innate immune response, including RIG-I-like receptor, toll-like receptor and NF-κB signaling pathway. Also, 12 down-regulated immune-related DEGs were selected for confirmation by reverse transcription quantitative real-time PCR verification, all these genes showed consistent expression between the result of reverse transcription quantitative real-time PCR and transcriptomic sequencing. These results revealed the important role of HSP70 in facilitating the innate immune response induced by TMUV infection. This is first to access the role of HSP70 in host response to TMUV infection, which provides a basis for further study of the pathogenesis of TMUV and contributes to the elucidation of TMUV-host interactions.

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转录组分析显示,坦布苏病毒感染诱导的热休克蛋白70 (HSP70)与免疫应答有关。
坦布苏病毒(TMUV)的爆发和流行给水禽养殖业带来了危害。然而,对TMUV感染的分子机制知之甚少。热休克蛋白70 (HSP70)是TMUV感染的正调控因子。为了研究HSP70与宿主对TMUV感染的免疫反应之间的相互作用,我们收集了携带或不携带HSP70抑制剂的TMUV感染细胞,并对其进行深度测序以鉴定差异表达的基因。我们在HSP70抑制剂处理和模拟处理的tmuv感染的DF-1细胞中发现了43个差异表达基因(DEGs)。在这些deg中,有39个基因显著下调。基因本体分析表明,这些基因主要参与生物过程、细胞成分和分子功能。京都基因与基因组百科分析显示,这些DEGs主要与先天免疫应答的激活有关,包括rig - i样受体、toll样受体和NF-κB信号通路。同时,选择12个下调的免疫相关deg进行反转录实时荧光定量PCR验证,这些基因在反转录实时荧光定量PCR结果和转录组测序结果之间表达一致。这些结果揭示了HSP70在促进TMUV感染诱导的先天免疫应答中的重要作用。这首次获得了HSP70在宿主对TMUV感染反应中的作用,为进一步研究TMUV的发病机制提供了基础,并有助于阐明TMUV-宿主相互作用。
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