Frontiers in Immunology (Jan 2020)

Isolation and Selection of Duck Primary Cells as Pathogenic and Innate Immunologic Cell Models for Duck Plague Virus

  • Bin Tian,
  • Bin Tian,
  • Bin Tian,
  • Dongjie Cai,
  • Tianqiong He,
  • Tianqiong He,
  • Tianqiong He,
  • Liyao Deng,
  • Liyao Deng,
  • Liyao Deng,
  • Liping Wu,
  • Liping Wu,
  • Liping Wu,
  • Mingshu Wang,
  • Mingshu Wang,
  • Mingshu Wang,
  • Renyong Jia,
  • Renyong Jia,
  • Renyong Jia,
  • Dekang Zhu,
  • Dekang Zhu,
  • Mafeng Liu,
  • Mafeng Liu,
  • Mafeng Liu,
  • Qiao Yang,
  • Qiao Yang,
  • Qiao Yang,
  • Ying Wu,
  • Ying Wu,
  • Ying Wu,
  • Xinxin Zhao,
  • Xinxin Zhao,
  • Xinxin Zhao,
  • Shun Chen,
  • Shun Chen,
  • Shun Chen,
  • Shaqiu Zhang,
  • Shaqiu Zhang,
  • Shaqiu Zhang,
  • Juan Huang,
  • Juan Huang,
  • Juan Huang,
  • Xumin Ou,
  • Xumin Ou,
  • Xumin Ou,
  • Sai Mao,
  • Sai Mao,
  • Sai Mao,
  • Yanling Yu,
  • Yanling Yu,
  • Yanling Yu,
  • Ling Zhang,
  • Ling Zhang,
  • Ling Zhang,
  • Yunya Liu,
  • Yunya Liu,
  • Yunya Liu,
  • Anchun Cheng,
  • Anchun Cheng,
  • Anchun Cheng

DOI
https://doi.org/10.3389/fimmu.2019.03131
Journal volume & issue
Vol. 10

Abstract

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Duck plague virus (DPV) is a representative pathogen transmitted among aquatic animals that causes gross lesions and immune inhibition in geese and ducks. The mechanism of organ tropism and innate immune evasion of DPV has not been completely deciphered due to a lack of cell models to study the innate immune manipulation and pathogenicity of aquatic viruses. In the present study, we isolated five types of duck primary cells [duck embryo fibroblasts (DEFs), neurons, astrocytes, peripheral blood mononuclear cells (PBMCs), and monocytes/macrophages] to identify appropriate cell models for DPV, using tropism infection and innate immunologic assays. Cells responded differently to stimulation with DNA viruses or RNA virus analogs. DPV infection exhibited broad tropism, as the recombinant virulent strain (CHv-GFP) infected DEFs, neurons, astrocytes, and monocytes/macrophages, but not the PBMCs, as the expression of EGFP was negligible. The basal levels of innate immunity molecules were highest in monocytes/macrophages and lower in DEFs and astrocytes. Conversely, the titer and genomic copy number of the attenuated virus strain was higher in DEFs and astrocytes than in neurons and monocytes/macrophages. The titer and genomic copy number of the attenuated virus strain were higher compared with the virulent strain in DEFs, neurons, and astrocytes. The innate immune response was not significantly induced by either DPV strain in DEFs, neurons, or astrocytes. The virulent strain persistently infected monocytes/macrophages, but the attenuated strain did so abortively, and this was accompanied by the phenomenon of innate immune inhibition and activation by the virulent and attenuated strains, respectively. Blockage of IFNAR signaling promoted replication of the attenuated strain. Pre-activation of IFNAR signaling inhibited infection by the virulent strain. The selection assay results indicated that induction of innate immunity plays an essential role in controlling DPV infection, and monocytes/macrophages are an important cell model for further investigations. Our study provided practical methods for isolating and culturing duck primary cells, and our results will facilitate further investigations of organ tropism, innate immune responses, latent infection, and the effectiveness of antiviral drugs for treating DPV and potentially other aerial bird pathogens.

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