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Recombinant influenza A virus hemagglutinin HA2 subunit protects mice against influenza A(H7N9) virus infection

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Abstract

A novel avian influenza A(H7N9) virus has emerged to infect humans in eastern China since 2013. An effective vaccine is needed because of the high mortality despite antiviral treatment and intensive care. We sought to develop an effective vaccine for A(H7N9) virus. The HA2 subunit was chosen as the vaccine antigen because it is highly conserved among the human A(H7N9) virus strains. Moreover, in silico analysis predicted two immunogenic regions within the HA2 subunit that may contain potential human B-cell epitopes. The HA2 fragment was readily expressed in Escherichia coli. In BALB/c mice, intraperitoneal immunization with two doses of HA2 with imiquimod (2-dose-imiquimod) elicited the highest geometric mean titer (GMT) of anti-HA2 IgG (12699), which was greater than that of two doses of HA2 without imiquimod (2-dose-no-adjuvant) (6350), one dose of HA2 with imiquimod (1-dose-imiquimod) (2000) and one dose of HA2 without imiquimod (1-dose-no-adjuvant) (794). The titer of anti-HA2 IgG was significantly higher in the 1-dose-imiquimod group than the 1-dose-no-adjuvant group. Although both hemagglutination inhibition titers and microneutralization titers were below 10, serum from immunized mice showed neutralizing activity in a fluorescent focus microneutralization assay. In a viral challenge experiment, the 2-dose-imiquimod group had the best survival rate (100 %), followed by the 2-dose-no-adjuvant group (90 %), the 1-dose-imiquimod group (70 %) and the 1-dose-no-adjuvant group (40 %). The 2-dose-imiquimod group also had significantly lower mean pulmonary viral loads than the 1-dose-imiquimod, 1-dose-no-adjuvant and non-immunized groups. This recombinant A(H7N9)-HA2 vaccine should be investigated as a complement to egg- or cell-based live attenuated or subunit influenza vaccines.

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Acknowledgments

We are grateful to Dr. Desmond Yap and Dr. Loretta Yuk-Yee Chan for performing the endotoxin assay. This work was supported in part by the Providence Foundation Ltd., in memory of the late Lui Hac Minh, and Mr. Larry Chi-Kin Yung. We thank the China Center for Disease Control and Prevention for providing the A/Anhui/1/2013 (H7N9) isolate. We acknowledge Yuelong Shu for his written permission to use his sequence data deposited in the Global Initiative on Sharing Avian Influenza Data (GISAID).

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The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. State Key Laboratory of Emerging Infectious Diseases, The University of Hong Kong, Hong Kong, China

      Kelvin K. W. To, Anna J. X. Zhang, Honglin Chen & Kwok-Yung Yuen

    2. Department of Microbiology, Carol Yu Centre for Infection, Queen Mary Hospital, The University of Hong Kong, Pokfulam Road, Pokfulam, Hong Kong, China

      Kelvin K. W. To, Honglin Chen & Kwok-Yung Yuen

    3. Research Centre of Infection and Immunology, The University of Hong Kong, Hong Kong, China

      Kelvin K. W. To, Anna J. X. Zhang, Honglin Chen & Kwok-Yung Yuen

    4. Department of Microbiology, The University of Hong Kong, Hong Kong, China

      Kelvin K. W. To, Anna J. X. Zhang, Andy S. F. Chan, Can Li, Jian-Piao Cai, Candy C. Y. Lau, Chuan-Gen Li, Akhee S. Jahan, Wai-Lan Wu, Alan Ka-Lun Tsang, Kwok-Hung Chan, Honglin Chen & Kwok-Yung Yuen

    5. Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Hangzhou, China

      Lanjuan Li, Honglin Chen & Kwok-Yung Yuen

    6. State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, College of Medicine, The First Affiliated Hospital, Zhejiang University, Hangzhou, China

      Lanjuan Li

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    1. Kelvin K. W. To
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    K. K. W. To and A. J. X. Zhang contributed equally to this wok.

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    To, K.K.W., Zhang, A.J.X., Chan, A.S.F. et al. Recombinant influenza A virus hemagglutinin HA2 subunit protects mice against influenza A(H7N9) virus infection. Arch Virol 160, 777–786 (2015). https://doi.org/10.1007/s00705-014-2314-x

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