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口蹄疫病毒感染后猪外周血单核细胞的转录组分析。

Transcriptomic analysis of porcine PBMCs in response to FMDV infection.

作者信息

Zhao Fu-Rong, Xie Yin-Li, Liu Ze-Zhong, Shao Jun-Jun, Li Shi-Fang, Zhang Yong-Guang, Chang Hui-Yun

机构信息

State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, People's Republic of China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu Province 225009, People's Republic of China; Key Laboratory of Fujian Province livestock epidemic prevention and control and biological technology, Longyan, Fujian Province 364012, People's Republic of China.

State Key Laboratory of Veterinary Etiological Biology, National Foot-and-Mouth Disease Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, People's Republic of China; Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, Jiangsu Province 225009, People's Republic of China.

出版信息

Acta Trop. 2017 Sep;173:69-75. doi: 10.1016/j.actatropica.2017.05.009. Epub 2017 May 8.

DOI:10.1016/j.actatropica.2017.05.009
PMID:28495401
Abstract

Foot-and-mouth disease (FMD) is a significant zoonotic infectious disease. It has an important economic impact throughout the world. As well, it is a considerable threat to food security. At present, the molecular mechanism of FMDV infection is not clear to a large extent. Innate immune response is the first line of defense against infectious diseases. The systematic analysis of the host immune response to infection has an important role in understanding the pathogenesis of infection. However, there are few reports about effect of immune regulation on virus replication in the interaction of virus and host cellular. High-throughput RNA-seq technology as a powerful and efficient means for transcript analysis provides a new insight into FMDV study. In this study, RNA extracted from pig PBMCs infected with O subtype FMDV at 4 dpi. A total of 29942658 and 31452917 Illumina read pairs were obtained from the non-infected (NI) group and infected (I) group, respectively. The clean bases for all samples are 3.61G (NI group) and 3.79G (I group), respectively. The clean reads of the NI and I group that mapped to pig genome data were 47195073 (81.82%) and 46556714 (76.85%), respectively. Most of the clean reads were distributed in the exon region, followed by intron region and intergenic region. Differently expressed (DE) genes were analyzed using edgeR software. 451 genes were differentially expressed between the infected and the non-infected groups. According to the comparison analysis, more genes were down-regulated in the non-infected samples than in those infected with FMDV.66 out of 451 genes were down-regulated, 385 out of 451 genes were up-regulated following FMDV infection. For function classification and pathway analysis, among 17741 assembled unigenes, there are 349 genes which are different genes of GO notes. Moreover, 49 genes were down-regulated, 300 genes were up-regulated associate with GO term. 1621 were successfully annotated by GO assignments, belonging to one or more of the three categories: biological process, cellular component, and molecular function. According to KEGG analysis,the main pathway was represented including protein processing in endoplasmic reticulum, phagosome, cell cycle and cytokine-cytokine receptor interaction. Some key DE genes related to immune process and signaling pathways were analyzed and quantified by RT-PCR. This is the first systematical transcriptome analysis of pig PBMCs infected by FMDV. These findings will help us better understand the host Cell-FMDV interaction and its relationship to pathogenesis, as well as contribute to the prevention and control of FMDV.

摘要

口蹄疫(FMD)是一种重要的人畜共患传染病。它在全球范围内具有重大的经济影响。此外,它对粮食安全构成了相当大的威胁。目前,口蹄疫病毒(FMDV)感染的分子机制在很大程度上尚不清楚。固有免疫反应是抵御传染病的第一道防线。对宿主感染免疫反应的系统分析对于理解感染的发病机制具有重要作用。然而,关于免疫调节在病毒与宿主细胞相互作用中对病毒复制影响的报道较少。高通量RNA测序技术作为一种强大而高效的转录本分析手段,为FMDV研究提供了新的视角。在本研究中,于感染后第4天从感染O型FMDV的猪外周血单核细胞(PBMCs)中提取RNA。未感染(NI)组和感染(I)组分别获得了总计29942658和31452917对Illumina读段。所有样本的净碱基分别为3.61G(NI组)和3.79G(I组)。映射到猪基因组数据的NI组和I组的净读段分别为47195073(81.82%)和46556714(76.85%)。大多数净读段分布在外显子区域,其次是内含子区域和基因间区域。使用edgeR软件分析差异表达(DE)基因。感染组和未感染组之间有451个基因差异表达。根据比较分析,未感染样本中下调的基因比感染FMDV的样本更多。451个基因中有66个下调,385个基因在FMDV感染后上调。对于功能分类和通路分析,在17741个组装的单基因中,有349个基因是具有不同GO注释的基因。此外,49个基因下调,300个基因上调与GO术语相关。1621个基因通过GO注释成功注释,属于生物过程、细胞成分和分子功能这三个类别中的一个或多个。根据KEGG分析,主要的通路包括内质网中的蛋白质加工、吞噬体、细胞周期和细胞因子 - 细胞因子受体相互作用。通过RT-PCR分析和定量了一些与免疫过程和信号通路相关的关键DE基因。这是首次对FMDV感染的猪PBMCs进行系统的转录组分析。这些发现将有助于我们更好地理解宿主细胞与FMDV的相互作用及其与发病机制的关系,也有助于FMDV的防控。

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