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Porcine epidemic diarrhea virus nsp4 induces pro-inflammatory cytokine and chemokine expression inhibiting viral replication in vitro

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Abstract

Porcine epidemic diarrhea virus (PEDV) causes severe economic loss in the pig industry each year. To better understand the relationship between cytokines and PEDV replication, in this study, pro-inflammatory cytokine and chemokine expression profiles in Vero cells infected with PEDV were analyzed. Real-time quantitative PCR assay indicated that IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 expression levels were significantly upregulated. Moreover, overexpression and siRNA silencing assays showed that overexpression of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 could significantly inhibit PEDV replication, while silencing of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 could significantly promote PEDV replication. Finally, a dual-luciferase reporter assay showed that nsp4 contributed to the expression of IL-1α, IL-1β, TNF-α, CCL2, CCL5 and CXCL8 via the NF-κB pathway. Together, these data determined that PEDV nsp4 could upregulate pro-inflammatory cytokine and chemokine expression, inhibiting viral replication in vitro. These results provided novel insights for understanding the roles of cytokines in PEDV replication.

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Abbreviations

PEDV:

Porcine epidemic diarrhea virus

ORFs:

Open reading frames

nsps:

Nonstructural proteins

S:

Spike protein

M:

Membrane protein

E:

Envelope protein

N:

Nucleocapsid protein

IFN:

Interferon

FBS:

Fetal bovine serum (Thermo)

PAMs:

Porcine alveolar macrophages

RT-PCR:

Reverse transcription polymerase chain reaction

RT-qPCR:

Real-time quantitative polymerase chain reaction

CT:

Threshold cycle

OD:

Optical density

SD:

Standard deviations

PBL:

Peripheral blood leucocytes

PCV2:

Porcine circovirus type 2

PBMCs:

Peripheral blood mononuclear cells

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Acknowledgements

This work was supported by the National Key Research and Development Program of China (2018YFD0501102), the National Key Technologies R&D Program (2015BAD12B02-5), the Guangzhou City Project (201508020062), and the Henan Science and Technology Project (182102110037).

Funding

This work was supported by the National Key Research and Development Program of China (2018YFD0501102), the National Key Technologies R&D Program (2015BAD12B02-5), the Guangzhou City Project (201508020062), and the Henan Science and Technology Project (182102110037).

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Author notes

  1. Linyang Yu, Jianguo Dong, and Yanwu Wang contributed equally to this work.

Authors and Affiliations

  1. Animal Science College/National Engineering Center for Swine Breeding Industry, South China Agriculture University, No. 483 Wushan Road, Tianhe District, Guangzhou, 510642, China

    Linyang Yu, Yanwu Wang, Pengfei Zhang, Yanling Liu, Leyi Zhang, Pengshuai Liang, Lei Wang & Changxu Song

  2. School of Animal Husbandry and Medical Engineering, Xinyang Agriculture and Forestry University, No. 1 North Road, Pingqiao District, Xinyang, 464000, China

    Jianguo Dong

  3. Guangxi State Farms Yongxin Animal Husbandry Group Co., Ltd, No. 135 Qixing Road, Qingxiu District, Nanning, 530000, China

    Yanwu Wang

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  1. Linyang Yu
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Contributions

Conceived and designed the experiments: Linyang Yu, Jianguo Dong, Changxu Song. Performed the experiments: Linyang Yu, Jianguo Dong, Yanwu Wang, Pengfei Zhang, Yanling Liu, Leyi Zhang, Pengshuai Liang, Lei Wang, Changxu Song. Analyzed the data and wrote the paper: Linyang Yu, Jianguo Dong, Changxu Song.

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Correspondence to Changxu Song.

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All of the samples were collected according to the animal ethics regulations of the National Engineering Center for Swine Breeding Industry (NECSBI 2015-16).

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Genetic data presented in this paper are publicly available via GenBank.

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Yu, L., Dong, J., Wang, Y. et al. Porcine epidemic diarrhea virus nsp4 induces pro-inflammatory cytokine and chemokine expression inhibiting viral replication in vitro. Arch Virol 164, 1147–1157 (2019). https://doi.org/10.1007/s00705-019-04176-2

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  • DOI: https://doi.org/10.1007/s00705-019-04176-2

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