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猪急性腹泻综合征冠状病毒感染期间IPEC-J2细胞基于RNA测序的全转录组分析

RNA-Seq-Based Whole Transcriptome Analysis of IPEC-J2 Cells During Swine Acute Diarrhea Syndrome Coronavirus Infection.

作者信息

Zhang Fanfan, Yuan Weifeng, Li Zhiquan, Zhang Yuhan, Ye Yu, Li Kai, Ding Zhen, Chen Yunyan, Cheng Ting, Wu Qiong, Tang Yuxin, Song Deping

机构信息

Key Laboratory for Animal Health of Jiangxi Province, Jiangxi Agricultural University, Nanchang, China.

Department of Preventive Veterinary Medicine, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang, China.

出版信息

Front Vet Sci. 2020 Aug 13;7:492. doi: 10.3389/fvets.2020.00492. eCollection 2020.

Abstract

The new emergence of swine acute diarrhea syndrome coronavirus (SADS-CoV) has resulted in high mortality in suckling pigs in China. To date, the transcriptional expression of host cells during SADS-CoV infection has not been documented. In this study, by means of RNA-Seq technology, we investigated the whole genomic expression profiles of intestinal porcine epithelial cells (IPEC-J2) infected with a SADS-CoV strain SADS-CoV-CH-FJWT-2018. A total of 24,676 genes were identified: 23,677 were known genes, and 999 were novel genes. A total of 1,897 differentially expressed genes (DEGs) were identified between SADS-CoV-infected and uninfected cells at 6, 24, and 48 h post infection (hpi). Of these, 1,260 genes were upregulated and 637 downregulated. A Gene Ontology enrichment analysis revealed that DEGs in samples from 6, 24, and 48 hpi were enriched in 79, 383, and 233 GO terms, respectively, which were mainly involved in immune system process, response to stimulus, signal transduction, and cytokine-cytokine receptor interactions. The 1,897 DEGs were mapped to 109 KEGG Ontology (KO) pathways classified into four main categories. Most of the DEGs annotated in the KEGG pathways were related to the immune system, infectious viral disease, and signal transduction. The mRNA of porcine serum amyloid A-3 protein (SAA3), an acute phase response protein, was significantly upregulated during the infection. Over-expressed SAA3 in IPEC-J2 cells drastically inhibited the replication of SADS-CoV, while under-expressed SAA3 promoted virus replication. To our knowledge, this is the first report on the profiles of gene expression of IPEC-J2 cells infected by SADS-CoV by means of RNA- technology. Our results indicate that SADS-CoV infection significantly modified the host cell gene expression patterns, and the host cells responded in highly specific manners, including immune response, signal and cytokine transduction, and antiviral response. The findings provide important insights into the transcriptome of IPEC-J2 in SADS-CoV infection.

摘要

猪急性腹泻综合征冠状病毒(SADS-CoV)的新出现导致中国哺乳仔猪的高死亡率。迄今为止,尚未有关于SADS-CoV感染期间宿主细胞转录表达的记录。在本研究中,我们借助RNA-Seq技术,研究了感染SADS-CoV毒株SADS-CoV-CH-FJWT-2018的猪肠上皮细胞(IPEC-J2)的全基因组表达谱。共鉴定出24,676个基因:其中23,677个为已知基因,999个为新基因。在感染后6、24和48小时(hpi),共鉴定出1,897个差异表达基因(DEG)。其中,1,260个基因上调,637个基因下调。基因本体富集分析显示,6、24和48 hpi样本中的DEG分别富集于79、383和233个基因本体(GO)术语中,主要涉及免疫系统过程、对刺激的反应、信号转导以及细胞因子-细胞因子受体相互作用。这1,897个DEG被映射到109条京都基因与基因组百科全书(KEGG)本体(KO)途径,分为四大类。KEGG途径中注释的大多数DEG与免疫系统、传染性病毒疾病和信号转导有关。急性期反应蛋白猪血清淀粉样蛋白A-3(SAA3)的mRNA在感染期间显著上调。在IPEC-J2细胞中过表达SAA3可显著抑制SADS-CoV的复制,而低表达SAA3则促进病毒复制。据我们所知,这是首次通过RNA技术报道SADS-CoV感染的IPEC-J2细胞的基因表达谱。我们的结果表明,SADS-CoV感染显著改变了宿主细胞基因表达模式,宿主细胞以高度特异性的方式做出反应,包括免疫反应、信号和细胞因子转导以及抗病毒反应。这些发现为SADS-CoV感染中IPEC-J2的转录组提供了重要见解。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cf49/7438718/2779fef7b409/fvets-07-00492-g0001.jpg

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