Phosphoproteomics to Characterize Host Response During H3N2 Canine Influenza Virus Infection of Dog Lung.

Yongbo Liu,Zhen Wang,Shiyu Tang,Shaotang Ye,Ying Chen,Cheng Fu,Yingxin Liang,Congwen Yao,Zhonghe Qi,Shoujun Li,Ji Wang,Siqi Cai

doi:10.3389/fvets.2020.585071

Abstract

Avian-origin H3N2 canine influenza viruses (CIVs) cause severe contagious respiratory disease in dogs, and quickly adapt to new environments. To further understand the mechanism of virus infection and host-virus interactions, we characterized the complete phosphoproteome of dogs infected with H3N2 CIV. Nine-week-old Beagle dogs were inoculated intranasally with 106 EID50 of A/canine/Guangdong/04/2014 (H3N2) virus. Lung sections were harvested at 5 days post-inoculation (dpi) and processed for global and quantitative analysis of differentially expressed phosphoproteins. A total of 1,235 differentially expressed phosphorylated proteins were identified in the dog lung after H3N2 CIV infection, and 3,016 modification sites were identified among all differentially expressed proteins. We then performed an enrichment analysis of functional annotations using Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) database analyses to predict the functions of the identified differential phosphoproteins. Our data indicate that H3N2 CIV infection causes dramatic changes in the host protein phosphorylation of dog lungs. To our knowledge, this is the first study to assess the effect of H3N2 CIV infection on the phosphoproteome of beagles. These data provide novel insights into H3N2-CIV-triggered regulatory phosphorylation circuits and signaling networks and may improve our understanding of the mechanisms underlying CIV pathogenesis in dogs.

Highlights

Influenza A is a highly contagious disease caused by influenza A virus (IAV) that has led to severe local and pandemic disease outbreaks, seriously threatening human and animal health [1]
In 2007, an outbreak of avian-origin H3N2 Canine influenza virus (CIV) was first reported in South Korea [14], which occurred via interspecies transmission from avian hosts to dogs [15]
We identified more than 1,200 host phosphoproteins that were induced or repressed in infected lungs, indicating that numerous definable phosphopeptides are regulated during CIV infection

Summary

Introduction

Influenza A is a highly contagious disease caused by influenza A virus (IAV) that has led to severe local and pandemic disease outbreaks, seriously threatening human and animal health [1]. Waterfowl are natural reservoir hosts for IAV, while the virus can be isolated from a wide variety of species, including humans, pigs, horses, and other mammals and birds. Phosphoproteomics to Characterize Canine Influenza and wide host spectrum are the main characteristics of IAVs, and the constant cross-species transmission is a constant threat to humans and other mammals [6, 7]. In 2007, an outbreak of avian-origin H3N2 CIV was first reported in South Korea [14], which occurred via interspecies transmission from avian hosts to dogs [15]. In 2015, an outbreak of H3N2 CIV originating from China occurred in the United States [17, 18], and more than 1,000 dogs were infected. The unique relationship between dogs and humans has raised concerns that CIVs may pose a potential risk to public health [22]

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