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      PB2-Q591K Mutation Determines the Pathogenicity of Avian H9N2 Influenza Viruses for Mammalian Species

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          Abstract

          Background

          Influenza A subtype H9N2 is widespread and prevalent in poultry. It has repeatedly transmitted zoonotically to cause mild influenza-like illness in humans and is regarded as a potential pandemic candidate. In additon, the six internal genes of H7N9 and H10N8 viruses which caused infection in human in China as well as some of the highly pathogenic H5N1 strains are origined from H9N2. Previous studies have shown that the mammalian adaptation PB2-Q591K contributes to the pathogenicity of H5N1 and H7N9 viruses. However, the role of the PB2-Q591K mutation in H9N2 subtype is still not well understood.

          Methods

          To define and compare the individual role of PB2-Q591K substitution in the PB2 gene segment of H9N2 in relation to polymerase activity, replication competence and the pathogenicity using in vitro and in vivo models.

          Results

          The PB2-Q591K mutation in H9N2 virus enhanced the polymerase activity and virus replication in human NHBE cells when compared to the wild type strain. Mice infected with the PB2 mutant showed significant weight loss, higher virus replication and immune responses in the lungs.

          Conclusions

          Our evidences suggest that the PB2-Q591K, in addition to the -E627K mutation in H9N2 enhanced the pathogenicity in mammalian host.

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          Most cited references18

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          Molecular basis for high virulence of Hong Kong H5N1 influenza A viruses.

          M Hatta (2001)
          In 1997, an H5N1 influenza A virus was transmitted from birds to humans in Hong Kong, killing 6 of the 18 people infected. When mice were infected with the human isolates, two virulence groups became apparent. Using reverse genetics, we showed that a mutation at position 627 in the PB2 protein influenced the outcome of infection in mice. Moreover, high cleavability of the hemagglutinin glycoprotein was an essential requirement for lethal infection.
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            Adaptive strategies of the influenza virus polymerase for replication in humans.

            Transmission of influenza viruses into the human population requires surmounting barriers to cross-species infection. Changes in the influenza polymerase overcome one such barrier. Viruses isolated from birds generally contain polymerases with the avian-signature glutamic acid at amino acid 627 in the PB2 subunit. These polymerases display restricted activity in human cells. An adaptive change in this residue from glutamic acid to the human-signature lysine confers high levels of polymerase activity in human cells. This mutation permits escape from a species-specific restriction factor that targets polymerases from avian viruses. A 2009 swine-origin H1N1 influenza A virus recently established a pandemic infection in humans, even though the virus encodes a PB2 with the restrictive glutamic acid at amino acid 627. We show here that the 2009 H1N1 virus has acquired second-site suppressor mutations in its PB2 polymerase subunit that convey enhanced polymerase activity in human cells. Introduction of this polymorphism into the PB2 subunit of a primary avian isolate also increased polymerase activity and viral replication in human and porcine cells. An alternate adaptive strategy has also been identified, whereby introduction of a human PA subunit into an avian polymerase overcomes restriction in human cells. These data reveal a strategy used by the 2009 H1N1 influenza A virus and identify other pathways by which avian and swine-origin viruses may evolve to enhance replication, and potentially pathogenesis, in humans.
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              Human infection with influenza H9N2.

              We report the clinical features of two cases of human infection with influenza A virus subtype H9N2 in Hong Kong, and show that serum samples from blood donors in Hong Kong had neutralising antibody suggestive of prior infection with influenza H9N2.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                29 September 2016
                2016
                : 11
                : 9
                : e0162163
                Affiliations
                [1 ]Department of Laboratory Medicine, Nanfang Hospital, Southern Medical University, GuangZhou, PR China
                [2 ]Centre of Influenza Research, School of Public Health, HKU Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, PR China
                [3 ]HKU-Pasteur Research Pole, School of Public Health, HKU Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong Special Administrative Region, PR China
                [4 ]State Key Laboratory of Respiratory Disease, National Clinical Research Center for Respiratory Disease, First Affiliated Hospital of Guangzhou Medical University, Guangzhou, PR China
                [5 ]Department of Laboratory Medicine, Guangdong No.2 Provincial People’s Hospital, GuangZhou, PR China
                Deutsches Primatenzentrum GmbH—Leibniz-Institut fur Primatenforschung, GERMANY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: CKPM ZZ.

                • Data curation: CKPM.

                • Formal analysis: CW CKPM ZFY ZZ.

                • Funding acquisition: CKPM ZFY.

                • Investigation: CW HHYL CKPM ZFY ZZ.

                • Methodology: CW HHYL CKPM.

                • Project administration: CW HHYL CKPM.

                • Resources: CKPM ZFY ZZ.

                • Software: CW HHYL.

                • Supervision: CKPM ZZ.

                • Validation: CW CKPM ZZ.

                • Visualization: CW HHYL.

                • Writing – original draft: CW CKPM ZZ.

                • Writing – review & editing: CW CKPM ZZ.

                Article
                PONE-D-16-17829
                10.1371/journal.pone.0162163
                5042486
                27684944
                53f6a4f8-3f84-41c3-b14b-b67506a6381f
                © 2016 Wang et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 9 May 2016
                : 11 August 2016
                Page count
                Figures: 7, Tables: 0, Pages: 16
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/501100005847, Health and Medical Research Fund;
                Award ID: 12111832
                Award Recipient :
                Funded by: municipal science and technology bureau foundation of guangzhou
                Award ID: 2014Y2-00031
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 81471937
                Award Recipient :
                This project was supported by Health and Medical Research Fund (no. 12111832), Municipal Science and Technology Bureau Foundation of Guangzhou (no. 2014Y2-00031) and National Natural Science Foundation of China (no. 81471937).
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