Table_1_Differential Transcriptomics Analysis of IPEC-J2 Cells Single or Coinfected With Porcine Epidemic Diarrhea Virus and Transmissible Gastroenteritis Virus.doc

Porcine epidemic diarrhea (PED) and transmissible gastroenteritis (TGE) caused by porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are two highly contagious intestinal diseases in the swine industry worldwide. Notably, coinfection of TGEV and PEDV is common in piglets with diarrhea-related diseases. In this study, intestinal porcine epithelial cells (IPEC-J2) were single or coinfected with PEDV and/or TGEV, followed by the comparison of differentially expressed genes (DEGs), especially interferon-stimulated genes (ISGs), between different groups via transcriptomics analysis and real-time qPCR. The antiviral activity of swine interferon-induced transmembrane protein 3 (sIFITM3) on PEDV and TGEV infection was also evaluated. The results showed that DEGs can be detected in the cells infected with PEDV, TGEV, and PEDV+TGEV at 12, 24, and 48 hpi, and the number of DEGs was the highest at 24 hpi. The DEGs are mainly annotated to the GO terms of protein binding, immune system process, organelle part, and intracellular organelle part. Furthermore, 90 ISGs were upregulated during PEDV or TGEV infection, 27 of which were associated with antiviral activity, including ISG15, OASL, IFITM1, and IFITM3. Furthermore, sIFITM3 can significantly inhibit PEDV and TGEV infection in porcine IPEC-J2 cells and/or monkey Vero cells. Besides, sIFITM3 can also inhibit vesicular stomatitis virus (VSV) replication in Vero cells. These results indicate that sIFITM3 has broad-spectrum antiviral activity.

由猪流行性腹泻病毒（Porcine epidemic diarrhea virus, PEDV）和传染性胃肠炎病毒（Transmissible gastroenteritis virus, TGEV）引发的猪流行性腹泻（Porcine epidemic diarrhea, PED）与传染性胃肠炎（Transmissible gastroenteritis, TGE），是全球养猪业中两种高传染性肠道疾病。值得注意的是，TGEV与PEDV的混合感染在出现腹泻相关病症的仔猪中极为常见。本研究中，研究人员将猪肠上皮细胞（Intestinal porcine epithelial cells, IPEC-J2）分别或联合用PEDV和/或TGEV进行感染，随后通过转录组学分析与实时定量聚合酶链反应（real-time qPCR），比较不同组别间的差异表达基因（Differentially expressed genes, DEGs），尤其是干扰素刺激基因（Interferon-stimulated genes, ISGs）的表达差异。本研究同时评估了猪干扰素诱导跨膜蛋白3（Swine interferon-induced transmembrane protein 3, sIFITM3）对PEDV和TGEV感染的抗病毒活性。结果显示，在感染后12、24、48小时（hours post infection, hpi），PEDV、TGEV以及PEDV与TGEV联合感染的细胞中均可检测到差异表达基因，且差异表达基因的数量在感染后24小时达到峰值。差异表达基因主要富集于基因本体（Gene Ontology, GO）注释条目：蛋白质结合、免疫系统过程、细胞器组分以及细胞内细胞器组分。此外，在PEDV或TGEV感染期间，共有90个干扰素刺激基因被上调，其中27个与抗病毒活性相关，包括ISG15、OASL、IFITM1以及IFITM3。进一步研究发现，sIFITM3可显著抑制猪IPEC-J2细胞以及（或）猴Vero细胞中的PEDV和TGEV感染。此外，sIFITM3还可抑制Vero细胞中的水泡性口炎病毒（Vesicular stomatitis virus, VSV）复制。上述结果表明，sIFITM3具有广谱抗病毒活性。