Replication of a Dog-Origin H6N1 influenza virus in cell culture and mice
Journal
Viruses
Journal Volume
12
Journal Issue
7
Date Issued
2020
Author(s)
Tsai, S.-K. et al.
Shih, C.-H.
Teng, K.-H.
Hsu, W.-E.
Lin, H.-J.
Huang, C.-H.
Abstract
The world's first natural avian-origin H6N1 influenza A virus infection case in dogs was confirmed in Taiwan in 2014. The H6N1 virus in chickens has been endemic in Taiwan since 1972. Whether the dog H6N1 virus has interspecies transmission potential is the key issue we aim to understand. Following one virus passage in embryonated eggs and two further passages in MDCK cells, we obtained two virus derivatives, E01EE (PB1 739E and PB2 627E) and E01GK (PB1 739G and PB2 627K), respectively. The pathogenicity of E01EE and E01GK was investigated using plaque assay, growth dynamic analysis and cell viability quantification in cells from different animal species. The impact of amino acid mutation on PB1 739 and PB2 627 on viral ribonucleoprotein (RNP) activity was also analyzed. Further mouse infection experiments were performed. The results showed that both E01EE and E01GK decreased cell relative viability of canine MDCK cells, human A549 cells and chicken DF1 cells. E01Gk caused greater cellular harm in MDCK and A549 cells and had significantly higher virus titers in all of the cells compared to E01EE. The PB2 627K but not PB1 739G was the critical mutation that influenced the viral RNP activity. Both E01EE and E01GK caused mice pneumonia and considerable virus shedding, especially E01GK. This report verifies PB2 E627K mutation in virulence and spotlights the potential for the dog H6N1 virus to extend interspecies transmission.
Other Subjects
amphotericin; chloramphenicol acetyltransferase; crystal violet; diaminobenzidine; eosin; formaldehyde; hematoxylin; isoflurane; penicillin derivative; peroxidase; ribonucleoprotein; streptomycin; trypsin; A-549 cell line; animal cell; animal model; animal tissue; antigen retrieval; Article; Bagg albino mouse; cell culture; cell viability assay; colorimetry; fetal calf serum; h6n1 influenza virus; histopathology; immunohistochemistry; Influenza virus; MDCK cell line; mouse; MTT assay; nonhuman; paraffin embedding; pathogenicity; plaque assay; real time reverse transcription polymerase chain reaction; tissue culture; virus infection; whole genome sequencing; animal; cell culture technique; dog; dog disease; genetics; growth, development and aging; human; Influenza A virus; mutation; orthomyxovirus infection; physiology; Taiwan; veterinary medicine; virology; virus replication; Animals; Cell Culture Techniques; Dog Diseases; Dogs; Humans; Influenza A virus; Madin Darby Canine Kidney Cells; Mice; Mice, Inbred BALB C; Mutation; Orthomyxoviridae Infections; Taiwan; Virus Replication
Type
journal article