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Generation and Characterization of Single-Cycle Infectious Canine Influenza A Virus (sciCIV) and Its Use as Vaccine Platform

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Vaccine Technologies for Veterinary Viral Diseases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2465))

Abstract

Influenza A viruses (IAVs) infect a broad range of hosts, including multiple avian and mammalian species. The frequent emergence of novel IAV strains in different hosts, including in humans, results in the need for vigilance and ongoing development of new approaches to fighting or prevent those infections. Canine influenza is a contagious respiratory disease in dogs caused by two subtypes of IAV, the equine-origin H3N8 canine influenza virus (CIV), and the avian-origin H3N2 CIV. A novel approach to influenza vaccination involves single-cycle infectious influenza A viruses (sciIAVs), which are defective for an essential viral gene. They are propagated in complementing cell lines which provide the missing gene in trans. As sciIAV cannot complete their replication cycle in regular cells they are limited to a single round of viral replication. Because of their safety profile and ability to express foreign antigens inside infected cells, sciIAVs have served both as live-attenuated vaccines and as vaccine vectors for the expression of heterologous antigens. Here, we describe experimental procedures for the generation of a single-cycle infectious CIV (sciCIV), where the viral hemagglutinin (HA) gene was exchanged for the gene for green fluorescent protein (GFP). Complementation of the viral HA protein is provided in trans by stable HA-expressing cell lines. Methods for the in vitro characterization of HA deficient but GFP-expressing sciCIV (sciCIV ΔHA/GFP) are described, as well as its use as a potential vaccine.

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Acknowledgments

We thank Biodefense and Emerging Infectious Research Resources Repository (BEI Resources) for providing polyclonal antibody against A/equine/Miami/1/1963 (H3N8) (BEI Resources NR-3103). This research was partially funded by the New York Influenza Center of Excellence (NYICE), a member of the National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), Department of Health and Human Services, Centers of Excellence for Influenza Research and Surveillance (CEIRS) contract No. HHSN272201400005C (NYICE). This research was partially funded by a “Ramon y Cajal” Incorporation grant (RYC-2017) from the Spanish Ministry of Science, Innovation and Universities to A.N.

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Authors and Affiliations

  1. Centro de Investigación en Sanidad Animal (CISA), INIA-CSIC, Madrid, Spain

    Aitor Nogales

  2. Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, NY, USA

    Aitor Nogales, Kevin Chiem, Michael Breen, Marta L. DeDiego & Luis Martínez-Sobrido

  3. Texas Biomedical Research Institute, San Antonio, TX, USA

    Kevin Chiem & Luis Martínez-Sobrido

  4. Boston University School of Medicine, Boston, MA, USA

    Michael Breen

  5. Centro Nacional de Biotecnología (CNB-CSIC), Madrid, Spain

    Marta L. DeDiego

  6. Department of Microbiology and Immunology, College of Veterinary Medicine, Baker Institute for Animal Health, Cornell University, Ithaca, NY, USA

    Colin R. Parrish

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  1. Aitor Nogales
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  2. Kevin Chiem
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  3. Michael Breen
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  4. Marta L. DeDiego
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  5. Colin R. Parrish
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  6. Luis Martínez-Sobrido
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Correspondence to Aitor Nogales or Luis Martínez-Sobrido .

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  1. Centro de Investigación en Sanidad Animal (CISA), INIA-CSIC, Valdeolmos, Madrid, Spain

    Alejandro Brun

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Nogales, A., Chiem, K., Breen, M., DeDiego, M.L., Parrish, C.R., Martínez-Sobrido, L. (2022). Generation and Characterization of Single-Cycle Infectious Canine Influenza A Virus (sciCIV) and Its Use as Vaccine Platform. In: Brun, A. (eds) Vaccine Technologies for Veterinary Viral Diseases. Methods in Molecular Biology, vol 2465. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2168-4_13

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  • DOI: https://doi.org/10.1007/978-1-0716-2168-4_13

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  • Publisher Name: Humana, New York, NY

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  • Online ISBN: 978-1-0716-2168-4

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