Abstract

The variant virulent porcine epidemic diarrhea virus (PEDV) strain (YN15) can cause severe porcine epidemic diarrhea (PED); however, the attenuated vaccine-like PEDV strain (YN144) can induce immunity in piglets. To investigate the differences in pathogenesis and epigenetic mechanisms between the two strains, differential expression and correlation analyses of the microRNA (miRNA) and mRNA in swine testicular (ST) cells infected with YN15, YN144, and mock were performed on three comparison groups (YN15 vs Control, YN144 vs Control, and YN15 vs YN144). The mRNA and miRNA expression profiles were obtained using next-generation sequencing (NGS), and the differentially expressed (DE) (p-value < 0.05) mRNA and miRNA were obtained using DESeq R package. mRNAs targeted by DE miRNAs were predicted using the miRanda algortithm. 8039, 8631 and 3310 DE mRNAs, and 36, 36, and 22 DE miRNAs were identified in the three comparison groups, respectively. 14,140, 15,367 and 3771 DE miRNA–mRNA (targeted by DE miRNAs) interaction pairs with negatively correlated expression patterns were identified, and interaction networks were constructed using Cytoscape. Six DE miRNAs and six DE mRNAs were randomly selected to verify the sequencing data by real-time relative quantitative reverse transcription polymerase chain reaction (qRT-PCR). Based on bioinformatics analysis, we discovered the differences were mostly involved in host immune responses and viral pathogenicity, including NF-κB signaling pathway and bacterial invasion of epithelial cells, etc. This is the first comprehensive comparison of DE miRNA–mRNA pairs in YN15 and YN144 infection in vitro, which could provide novel strategies for the prevention and control of PED.

Highlights

  • The variant virulent porcine epidemic diarrhea virus (PEDV) strain (YN15) can cause severe porcine epidemic diarrhea (PED); the attenuated vaccine-like PEDV strain (YN144) can induce immunity in piglets

  • The differential expression and correlation analysis of the differentially expressed (DE, p-value < 0.05) miRNA and mRNA in swine testicular (ST) cells infected with the variant virulent PEDV strain (YN15) and the attenuated vaccine-like PEDV strain (YN144) were investigated to understand the roles of the miRNA–mRNA network in PEDV pathogenesis using next-generation sequencing (NGS) technology

  • In view of the significant differences in pathogenesis and epigenetic mechanisms between the variant virulent PEDV strain YN15 and the attenuated vaccine-like PEDV strain YN144 in vivo, we suppose that miRNA and mRNA may play essential roles in PEDV virulence

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Summary

Introduction

The variant virulent porcine epidemic diarrhea virus (PEDV) strain (YN15) can cause severe porcine epidemic diarrhea (PED); the attenuated vaccine-like PEDV strain (YN144) can induce immunity in piglets. We discovered the differences were mostly involved in host immune responses and viral pathogenicity, including NF-κB signaling pathway and bacterial invasion of epithelial cells, etc. This is the first comprehensive comparison of DE miRNA–mRNA pairs in YN15 and YN144 infection in vitro, which could provide novel strategies for the prevention and control of PED. In order to better understand the pathogenic mechanism of the variant PEDVs, a proteomics study was performed in vivo following the variant and attenuated strain infection, whose differentially expressed proteins are associated with immune function and stress r­ esponses[6]. Next-generation sequencing (NGS) has become a powerful tool to analyze the expression profiles of miRNA and ­mRNA12

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