Plasticity of the influenza virus H5 HA protein

Huihui Kong; David F. Burke; Tiago Jose da Silva Lopes; Kosuke Takada; Masaki Imai; Gongxun Zhong; Masato Hatta; Shufang Fan; Shiho Chiba; Derek Smith; Gabriele Neumann; Yoshihiro Kawaoka

doi:10.1128/mBio.03324-20

Plasticity of the influenza virus H5 HA protein

Huihui Kong, David F. Burke, Tiago Jose da Silva Lopes, Kosuke Takada, Masaki Imai, Gongxun Zhong, Masato Hatta, Shufang Fan, Shiho Chiba, Derek Smith, Gabriele Neumann, Yoshihiro Kawaoka^*

^*Corresponding author for this work

Centre for Host Microbiome Interactions

Research output: Contribution to journal › Article › peer-review

9 Citations (Scopus)

Abstract

Since the emergence of highly pathogenic avian influenza viruses of the H5 subtype, the major viral antigen, hemagglutinin (HA), has undergone con-stant evolution, resulting in numerous genetic and antigenic (sub)clades. To explore the consequences of amino acid changes at sites that may affect the antigenicity of H5 viruses, we simultaneously mutated 17 amino acid positions of an H5 HA by using a synthetic gene library that, theoretically, encodes all combinations of the 20 amino acids at the 17 positions. All 251 mutant viruses sequenced possessed ≥13 amino acid substitutions in HA, demonstrating that the targeted sites can accommo-date a substantial number of mutations. Selection with ferret sera raised against H5 viruses of different clades resulted in the isolation of 39 genotypes. Further analysis of seven variants demonstrated that they were antigenically different from the parental virus and replicated efficiently in mammalian cells. Our data demonstrate the substantial plasticity of the influenza virus H5 HA protein, which may lead to novel antigenic variants. IMPORTANCE The HA protein of influenza A viruses is the major viral antigen. In this study, we simultaneously introduced mutations at 17 amino acid positions of an H5 HA expected to affect antigenicity. Viruses with ≥13 amino acid changes in HA were viable, and some had altered antigenic properties. H5 HA can therefore accommo-date many mutations in regions that affect antigenicity. The substantial plasticity of H5 HA may facilitate the emergence of novel antigenic variants.

Original language	English
Article number	e03324-20
Pages (from-to)	1-14
Number of pages	14
Journal	Mbio
Volume	12
Issue number	1
DOIs	https://doi.org/10.1128/mBio.03324-20
Publication status	Published - 2021

Keywords

HA
Influenza
Sequence plasticity

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10.1128/mBio.03324-20

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title = "Plasticity of the influenza virus H5 HA protein",

abstract = "Since the emergence of highly pathogenic avian influenza viruses of the H5 subtype, the major viral antigen, hemagglutinin (HA), has undergone con-stant evolution, resulting in numerous genetic and antigenic (sub)clades. To explore the consequences of amino acid changes at sites that may affect the antigenicity of H5 viruses, we simultaneously mutated 17 amino acid positions of an H5 HA by using a synthetic gene library that, theoretically, encodes all combinations of the 20 amino acids at the 17 positions. All 251 mutant viruses sequenced possessed ≥13 amino acid substitutions in HA, demonstrating that the targeted sites can accommo-date a substantial number of mutations. Selection with ferret sera raised against H5 viruses of different clades resulted in the isolation of 39 genotypes. Further analysis of seven variants demonstrated that they were antigenically different from the parental virus and replicated efficiently in mammalian cells. Our data demonstrate the substantial plasticity of the influenza virus H5 HA protein, which may lead to novel antigenic variants. IMPORTANCE The HA protein of influenza A viruses is the major viral antigen. In this study, we simultaneously introduced mutations at 17 amino acid positions of an H5 HA expected to affect antigenicity. Viruses with ≥13 amino acid changes in HA were viable, and some had altered antigenic properties. H5 HA can therefore accommo-date many mutations in regions that affect antigenicity. The substantial plasticity of H5 HA may facilitate the emergence of novel antigenic variants.",

keywords = "HA, Influenza, Sequence plasticity",

author = "Huihui Kong and Burke, {David F.} and Lopes, {Tiago Jose da Silva} and Kosuke Takada and Masaki Imai and Gongxun Zhong and Masato Hatta and Shufang Fan and Shiho Chiba and Derek Smith and Gabriele Neumann and Yoshihiro Kawaoka",

note = "Funding Information: This project has been funded in part with federal funds from the Department of Health and Human Services, Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority, under contract no. HHSO100201500033C. This study was also supported by the NIAID-funded Center for Research on Influenza Pathogenesis (CRIP; HHSN272201400008C), by the Research Program on Emerging and Re-emerging Infectious Diseases from the Japan Agency for Medical Research and Development (AMED; JP19fk0108031), by the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) from the AMED (JP19fm0108006), by the Japan Program for Infectious Diseases Research and Infrastructure from the AMED (JP20wm0125002), and by Grants-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan (no. 16H06429, 16K21723, and 16H06434). Publisher Copyright: {\textcopyright} 2021 Kong et al.",

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doi = "10.1128/mBio.03324-20",

language = "English",

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T1 - Plasticity of the influenza virus H5 HA protein

AU - Kong, Huihui

AU - Burke, David F.

AU - Lopes, Tiago Jose da Silva

AU - Takada, Kosuke

AU - Imai, Masaki

AU - Zhong, Gongxun

AU - Hatta, Masato

AU - Fan, Shufang

AU - Chiba, Shiho

AU - Smith, Derek

AU - Neumann, Gabriele

AU - Kawaoka, Yoshihiro

N1 - Funding Information: This project has been funded in part with federal funds from the Department of Health and Human Services, Office of the Assistant Secretary for Preparedness and Response, Biomedical Advanced Research and Development Authority, under contract no. HHSO100201500033C. This study was also supported by the NIAID-funded Center for Research on Influenza Pathogenesis (CRIP; HHSN272201400008C), by the Research Program on Emerging and Re-emerging Infectious Diseases from the Japan Agency for Medical Research and Development (AMED; JP19fk0108031), by the Japan Initiative for Global Research Network on Infectious Diseases (J-GRID) from the AMED (JP19fm0108006), by the Japan Program for Infectious Diseases Research and Infrastructure from the AMED (JP20wm0125002), and by Grants-in-Aid for Scientific Research on Innovative Areas from the Ministry of Education, Culture, Science, Sports, and Technology (MEXT) of Japan (no. 16H06429, 16K21723, and 16H06434). Publisher Copyright: © 2021 Kong et al.

PY - 2021

Y1 - 2021

N2 - Since the emergence of highly pathogenic avian influenza viruses of the H5 subtype, the major viral antigen, hemagglutinin (HA), has undergone con-stant evolution, resulting in numerous genetic and antigenic (sub)clades. To explore the consequences of amino acid changes at sites that may affect the antigenicity of H5 viruses, we simultaneously mutated 17 amino acid positions of an H5 HA by using a synthetic gene library that, theoretically, encodes all combinations of the 20 amino acids at the 17 positions. All 251 mutant viruses sequenced possessed ≥13 amino acid substitutions in HA, demonstrating that the targeted sites can accommo-date a substantial number of mutations. Selection with ferret sera raised against H5 viruses of different clades resulted in the isolation of 39 genotypes. Further analysis of seven variants demonstrated that they were antigenically different from the parental virus and replicated efficiently in mammalian cells. Our data demonstrate the substantial plasticity of the influenza virus H5 HA protein, which may lead to novel antigenic variants. IMPORTANCE The HA protein of influenza A viruses is the major viral antigen. In this study, we simultaneously introduced mutations at 17 amino acid positions of an H5 HA expected to affect antigenicity. Viruses with ≥13 amino acid changes in HA were viable, and some had altered antigenic properties. H5 HA can therefore accommo-date many mutations in regions that affect antigenicity. The substantial plasticity of H5 HA may facilitate the emergence of novel antigenic variants.

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Plasticity of the influenza virus H5 HA protein

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