Transcriptome analysis of host response to porcine epidemic diarrhea virus nsp15 in IPEC-J2 cells

Tao Ouyang; Zibing Yang; Jiawu Wan; Yanni Zhang; Xiaoling Wang; Lingbao Kong; Ting Wang; Yihan Li

doi:10.1016/j.micpath.2021.105195

Transcriptome analysis of host response to porcine epidemic diarrhea virus nsp15 in IPEC-J2 cells

Microb Pathog. 2022 Jan:162:105195. doi: 10.1016/j.micpath.2021.105195. Epub 2021 Sep 24.

Authors

Tao Ouyang¹, Zibing Yang¹, Jiawu Wan¹, Yanni Zhang², Xiaoling Wang¹, Lingbao Kong¹, Ting Wang³, Yihan Li⁴

Affiliations

¹ Institute of Pathogenic Microorganism and College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang, Jiangxi, China.
² Jiangxi Province Center for Disease Control and Prevention, Nanchang, Jiangxi, China.
³ Institute of Pathogenic Microorganism and College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang, Jiangxi, China. Electronic address: tingwang@jxau.edu.cn.
⁴ Institute of Pathogenic Microorganism and College of Bioscience and Engineering, Jiangxi Agricultural University, Nanchang, Jiangxi, China. Electronic address: liyihan@jxau.edu.cn.

PMID: 34571150
DOI: 10.1016/j.micpath.2021.105195

Abstract

Background: Porcine epidemic diarrhea virus (PEDV) is an enveloped positive-sense ssRNA virus which is highly lethal to piglets, causing enormous economic losses to swine industry worldwide. Nsp15 protein is an endoribonuclease of PEDV and plays an indispensable role in the viral proliferation. We reported the transcription files of nsp15 transfected IPEC-J2 cells for the first time to broaden our understanding of PEDV pathogenesis.

Methods: RNA-seq was performed to compare gene expression profiles between pCAGGS-HA-nsp15 transfected IPEC-J2 cells and pCAGGS-HA (empty vector) transfected IPEC-J2 cells. Immune-related genes and pathways were identified and analyzed to deepen our understanding of nsp15 for PEDV pathogenicity. IPEC-J2 cells transfected with pCAGGS-HA-CCL5/CXCL8/CXCL10 were infected with CV777 and the virus load of PEDV was detected by qRT-PCR.

Results: A total of 21,654 genes were obtained by RNA-Seq and 415 differential expressed genes (DEGs) were identified, including 136 up-regulated and 279 down-regulated genes. A number of effect genes involved in immune responses and inflammation were differentially expressed. GO and KEGG enrichment analysis showed that 32 GO terms were significantly enriched and the DEGs were mainly enriched in immune-related pathways such as TNF signaling pathway, RIG-I-like receptor signaling pathway and Cytosolic DNA-sensing pathway. qRT-PCR results indicated the overexpression of selected chemokines, CCL5/CXCL8/CXCL10, can inhibit PEDV proliferation in IPEC-J2 cells.

Conclusions: Our transcriptome profile illustrated a number of genes involving in immune responses and inflammation were inhibited by nsp15, such as CCL5, CXCL8, CXCL10, OAS, MXs, STAT1 and IRF9. The results suggested that nsp15 can antagonize IFNs and block chemokine system to provide an adequate intracellular environment for viral proliferation.

Keywords: Chemokines; Gene expression; Transcriptome; nsp15.

MeSH terms

Animals
Cell Line
Coronavirus Infections* / genetics
Coronavirus Infections* / veterinary
Epithelial Cells
Gene Expression Profiling
Immunity
Porcine epidemic diarrhea virus* / genetics
Swine