Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection

Abstract The global spread, frequent antigenic changes, and pandemic potential of clade 2.3.4.4b highly pathogenic avian influenza H5N1 underscore the urgent need for robust cross-protective vaccines. Here, we developed a clade 2.3.4.4b H5N1 whole inactivated virus (WIV) vaccine strain with improved...

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Main Authors: Jin-Ha Song, Seung-Eun Son, Ho-Won Kim, Seung-Ji Kim, Se-Hee An, Chung-Young Lee, Hyuk-Joon Kwon, Kang-Seuk Choi
Format: Article
Language:English
Published: BMC 2025-05-01
Series:Virology Journal
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Online Access:https://doi.org/10.1186/s12985-025-02760-4
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author Jin-Ha Song
Seung-Eun Son
Ho-Won Kim
Seung-Ji Kim
Se-Hee An
Chung-Young Lee
Hyuk-Joon Kwon
Kang-Seuk Choi
author_facet Jin-Ha Song
Seung-Eun Son
Ho-Won Kim
Seung-Ji Kim
Se-Hee An
Chung-Young Lee
Hyuk-Joon Kwon
Kang-Seuk Choi
author_sort Jin-Ha Song
collection DOAJ
description Abstract The global spread, frequent antigenic changes, and pandemic potential of clade 2.3.4.4b highly pathogenic avian influenza H5N1 underscore the urgent need for robust cross-protective vaccines. Here, we developed a clade 2.3.4.4b H5N1 whole inactivated virus (WIV) vaccine strain with improved structural stability, productivity, and safety. By analyzing the evolutionary trends of clade 2.3.4.4b H5N1 viruses, we identified a key mutation (R90K) that increases heat stability while preserving antigenicity. Additionally, the PB2 gene of PR8 was replaced with a prototypical avian PB2 gene to increase replication efficiency in embryonated chicken eggs and reduce replication efficiency in mammalian cells, thereby improving productivity and biosafety. We found that our optimized clade 2.3.4.4b H5N1 vaccine strain (22W_KY), inactivated with binary ethylenimine (BEI), had superior antigen internalization into respiratory epithelial cells compared to those inactivated with formaldehyde or beta-propiolactone. Following intranasal administration to mice, the BEI-inactivated 22W_KY also elicited significantly stronger systemic IgG, mucosal IgA, and T-cell responses, especially in the lungs. Protective efficacy studies revealed that the BEI-inactivated 22W_KY vaccine provided complete protection against heterologous viral challenges and significant protection against heterosubtypic viral challenges, with no weight loss and complete suppression of the viral load in the respiratory tract in 2 of 3 mice. These results indicate that the BEI-inactivated 22W_KY vaccine could serve as a promising candidate for a safe, stable, cost-efficient, and broadly protective intranasal influenza vaccine against zoonotic and pandemic threats.
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spelling doaj-art-2dd70f9791e8406f978e0ee89b5d1dc02025-08-20T01:51:59ZengBMCVirology Journal1743-422X2025-05-0122112510.1186/s12985-025-02760-4Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protectionJin-Ha Song0Seung-Eun Son1Ho-Won Kim2Seung-Ji Kim3Se-Hee An4Chung-Young Lee5Hyuk-Joon Kwon6Kang-Seuk Choi7Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National UniversityLaboratory of Avian Diseases, College of Veterinary Medicine, Seoul National UniversityLaboratory of Avian Diseases, College of Veterinary Medicine, Seoul National UniversityLaboratory of Avian Diseases, College of Veterinary Medicine, Seoul National UniversityAvian Influenza Research & Diagnostic Division, Animal and Plant Quarantine AgencyDepartment of Microbiology, College of Medicine, Kyungpook National UniversityLaboratory of Poultry Medicine, Department of Farm Animal Medicine, College of Veterinary Medicine, BK21 PLUS for Veterinary Science, Seoul National UniversityLaboratory of Avian Diseases, College of Veterinary Medicine, Seoul National UniversityAbstract The global spread, frequent antigenic changes, and pandemic potential of clade 2.3.4.4b highly pathogenic avian influenza H5N1 underscore the urgent need for robust cross-protective vaccines. Here, we developed a clade 2.3.4.4b H5N1 whole inactivated virus (WIV) vaccine strain with improved structural stability, productivity, and safety. By analyzing the evolutionary trends of clade 2.3.4.4b H5N1 viruses, we identified a key mutation (R90K) that increases heat stability while preserving antigenicity. Additionally, the PB2 gene of PR8 was replaced with a prototypical avian PB2 gene to increase replication efficiency in embryonated chicken eggs and reduce replication efficiency in mammalian cells, thereby improving productivity and biosafety. We found that our optimized clade 2.3.4.4b H5N1 vaccine strain (22W_KY), inactivated with binary ethylenimine (BEI), had superior antigen internalization into respiratory epithelial cells compared to those inactivated with formaldehyde or beta-propiolactone. Following intranasal administration to mice, the BEI-inactivated 22W_KY also elicited significantly stronger systemic IgG, mucosal IgA, and T-cell responses, especially in the lungs. Protective efficacy studies revealed that the BEI-inactivated 22W_KY vaccine provided complete protection against heterologous viral challenges and significant protection against heterosubtypic viral challenges, with no weight loss and complete suppression of the viral load in the respiratory tract in 2 of 3 mice. These results indicate that the BEI-inactivated 22W_KY vaccine could serve as a promising candidate for a safe, stable, cost-efficient, and broadly protective intranasal influenza vaccine against zoonotic and pandemic threats.https://doi.org/10.1186/s12985-025-02760-4Highly pathogenic avian influenza virusHeat stabilityWhole inactivated virus vaccineBinary ethylenimineIntranasal administrationCross-protection
spellingShingle Jin-Ha Song
Seung-Eun Son
Ho-Won Kim
Seung-Ji Kim
Se-Hee An
Chung-Young Lee
Hyuk-Joon Kwon
Kang-Seuk Choi
Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection
Virology Journal
Highly pathogenic avian influenza virus
Heat stability
Whole inactivated virus vaccine
Binary ethylenimine
Intranasal administration
Cross-protection
title Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection
title_full Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection
title_fullStr Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection
title_full_unstemmed Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection
title_short Intranasally administered whole virion inactivated vaccine against clade 2.3.4.4b H5N1 influenza virus with optimized antigen and increased cross-protection
title_sort intranasally administered whole virion inactivated vaccine against clade 2 3 4 4b h5n1 influenza virus with optimized antigen and increased cross protection
topic Highly pathogenic avian influenza virus
Heat stability
Whole inactivated virus vaccine
Binary ethylenimine
Intranasal administration
Cross-protection
url https://doi.org/10.1186/s12985-025-02760-4
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