{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:42:01Z","timestamp":1760146921906,"version":"build-2065373602"},"reference-count":61,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2024,12,30]],"date-time":"2024-12-30T00:00:00Z","timestamp":1735516800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003725","name":"National Research Foundation of Korea","doi-asserted-by":"publisher","award":["NRF-2022M3A9I2017587"],"award-info":[{"award-number":["NRF-2022M3A9I2017587"]}],"id":[{"id":"10.13039\/501100003725","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background\/Objective: Highly pathogenic (HP) H5Nx and low-pathogenicity (LP) H9N2 avian influenza viruses (AIVs) pose global threats to the poultry industry and public health, highlighting the critical need for a dual-protective vaccine. Methods: In this study, we generated a model PR8-derived recombinant H5N2 vaccine strain with hemagglutinin (HA) and neuraminidase (NA) genes from clade 2.3.2.1c H5N1 and Y439-like H9N2 viruses, respectively. To enhance the immunogenicity of the recombinant H5N2 vaccine strain, N-glycans of the HA2 subunit, NA, and M2e were modified. Additionally, we replaced M2e with avian M2e to enhance the antigenic homogeneity of AIVs for better protection. We also replaced PR8 PB2 with 01310 PB2, which is the PB2 gene derived from an LP H9N2 avian influenza virus, to eliminate pathogenicity in mammals. The productivity of the model vaccine strain (rvH5N2-aM2e-vPB2) in embryonated chicken eggs (ECEs), its potential risk of mammalian infection, and the immunogenicity associated with different inactivation methods (formaldehyde (F\/A) vs. binary ethyleneimine (BEI)) were evaluated. Results: The rvH5N2-aM2e-vPB2 strain demonstrated high productivity in ECEs and exhibited complete inhibition of replication in mammalian cells. Furthermore, compared with using F\/A inactivation, inactivation using BEI significantly enhanced the immune response, particularly against NA. This enhancement resulted in increased virus neutralization titers, supporting its efficacy for dual protection against H5Nx and H9N2 avian influenza viruses. Furthermore, we demonstrated that M2e-specific immune responses, difficult to induce with inactivated vaccines, can be effectively elicited with live vaccines, suggesting a strategy to enhance M2e immunogenicity in whole influenza virus vaccines. Conclusions: Finally, the successful development of the model rH5N2 vaccine strain is described; this strain provides dual protection, has potential applicability in regions where avian influenza is endemic, and can be used to promote the development of versatile H5N2 recombinant vaccines for effective avian influenza control.<\/jats:p>","DOI":"10.3390\/vaccines13010022","type":"journal-article","created":{"date-parts":[[2024,12,31]],"date-time":"2024-12-31T07:51:57Z","timestamp":1735631517000},"page":"22","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Model H5N2 Vaccine Strain for Dual Protection Against H5N1 and H9N2 Avian Influenza Viruses"],"prefix":"10.3390","volume":"13","author":[{"given":"Jin-Ha","family":"Song","sequence":"first","affiliation":[{"name":"Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seung-Eun","family":"Son","sequence":"additional","affiliation":[{"name":"Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5509-1152","authenticated-orcid":false,"given":"Ho-Won","family":"Kim","sequence":"additional","affiliation":[{"name":"Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2206-1299","authenticated-orcid":false,"given":"Se-Hee","family":"An","sequence":"additional","affiliation":[{"name":"Avian Influenza Research & Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon-si 39660, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3037-7581","authenticated-orcid":false,"given":"Chung-Young","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Microbiology, College of Medicine, Kyungpook National University, Daegu 41944, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9107-7860","authenticated-orcid":false,"given":"Hyuk-Joon","family":"Kwon","sequence":"additional","affiliation":[{"name":"Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea"},{"name":"Laboratory of Poultry Medicine, Department of Farm Animal Medicine, College of Veterinary Medicine and BK21 PLUS for Veterinary Science, Seoul National University, Seoul 88026, Republic of Korea"},{"name":"Farm Animal Clinical Training and Research Center (FACTRC), Institutes of Green Bio Science and Technology (GBST), Seoul National University, Pyeongchang 25354, Republic of Korea"},{"name":"GeNiner Inc., Seoul 08826, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6825-6924","authenticated-orcid":false,"given":"Kang-Seuk","family":"Choi","sequence":"additional","affiliation":[{"name":"Laboratory of Avian Diseases, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea"},{"name":"Research Institute for Veterinary Science, College of Veterinary Medicine, Seoul National University, Seoul 08826, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2024,12,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"810","DOI":"10.1038\/s41586-023-06631-2","article-title":"The episodic resurgence of highly pathogenic avian influenza H5 virus","volume":"622","author":"Xie","year":"2023","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"278","DOI":"10.1016\/j.meegid.2015.06.004","article-title":"Evolutionary trajectories and diagnostic challenges of potentially zoonotic avian influenza viruses H5N1 and H9N2 co-circulating in Egypt","volume":"34","author":"Naguib","year":"2015","journal-title":"Infect. Genet. Evol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"5760","DOI":"10.1128\/JCM.43.11.5760-5767.2005","article-title":"Human Infection with an Avian H9N2 Influenza A Virus in Hong Kong in 2003","volume":"43","author":"Butt","year":"2005","journal-title":"J. Clin. Microbiol."},{"key":"ref_4","first-page":"2300001","article-title":"Highly pathogenic avian influenza A(H5N1) virus infection in farmed minks, Spain, October 2022","volume":"28","author":"Monne","year":"2023","journal-title":"Eurosurveillance"},{"key":"ref_5","unstructured":"(2024, March 03). Reported Human Infections with Avian Influenza A Viruses|Avian Influenza (Flu), Available online: https:\/\/www.cdc.gov\/bird-flu\/php\/avian-flu-summary\/reported-human-infections.html."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/s41426-018-0029-x","article-title":"Comparison of the pathogenic potential of highly pathogenic avian influenza (HPAI) H5N6, and H5N8 viruses isolated in South Korea during the 2016\u20132017 winter season","volume":"7","author":"Kwon","year":"2018","journal-title":"Emerg. Microbes Infect."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e3255","DOI":"10.1111\/tbed.14551","article-title":"Emergence of clade 2.3.4.4b novel reassortant H5N1 high pathogenicity avian influenza virus in South Korea during late 2021","volume":"69","author":"Sagong","year":"2022","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"e5","DOI":"10.4142\/jvs.22216","article-title":"Current situation and control strategies of H9N2 avian influenza in South Korea","volume":"24","author":"Sagong","year":"2022","journal-title":"J. Vet. Sci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"10-1128","DOI":"10.1128\/mBio.02332-17","article-title":"NAction! How Can Neuraminidase-Based Immunity Contribute to Better Influenza Virus Vaccines?","volume":"9","author":"Krammer","year":"2018","journal-title":"mBio"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"S75","DOI":"10.1093\/infdis\/jiz017","article-title":"Neuraminidase, the Forgotten Surface Antigen, Emerges as an Influenza Vaccine Target for Broadened Protection","volume":"219","author":"Eichelberger","year":"2019","journal-title":"J. Infect. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"10-1128","DOI":"10.1128\/mBio.02556-14","article-title":"Vaccination with Adjuvanted Recombinant Neuraminidase Induces Broad Heterologous, but Not Heterosubtypic, Cross-Protection against Influenza Virus Infection in Mice","volume":"6","author":"Wohlbold","year":"2015","journal-title":"mBio"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Kim, K.-H., Li, Z., Bhatnagar, N., Subbiah, J., Park, B.R., Shin, C.H., Pushko, P., Wang, B.-Z., and Kang, S.-M. (2022). Universal protection against influenza viruses by multi-subtype neuraminidase and M2 ectodomain virus-like particle. PLoS Pathog., 18.","DOI":"10.1371\/journal.ppat.1010755"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2820","DOI":"10.1128\/JVI.03246-14","article-title":"A viable recombinant rhabdovirus lacking its glycoprotein gene and expressing influenza virus hemagglutinin and neuraminidase is a potent influenza vaccine","volume":"89","author":"Ryder","year":"2015","journal-title":"J. Virol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"3175","DOI":"10.1016\/j.vaccine.2004.01.055","article-title":"Generation of reassortant influenza vaccines by reverse genetics that allows utilization of a DIVA (Differentiating Infected from Vaccinated Animals) strategy for the control of avian influenza","volume":"22","author":"Lee","year":"2004","journal-title":"Vaccine"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"18137","DOI":"10.1073\/pnas.0909696106","article-title":"Glycans on influenza hemagglutinin affect receptor binding and immune response","volume":"106","author":"Wang","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Li, J., Dohna, H.z., Cardona, C.J., Miller, J., and Carpenter, T.E. (2011). Emergence and genetic variation of neuraminidase stalk deletions in avian influenza viruses. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0014722"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"299","DOI":"10.4142\/jvs.2017.18.S1.299","article-title":"Optimized clade 2.3. 2.1 c H5N1 recombinant-vaccine strains against highly pathogenic avian influenza","volume":"18","author":"Jang","year":"2017","journal-title":"J. Vet. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"67","DOI":"10.4142\/jvs.2008.9.1.67","article-title":"An inactivated vaccine to control the current H9N2 low pathogenic avian influenza in Korea","volume":"9","author":"Choi","year":"2008","journal-title":"J. Vet. Sci."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1016\/j.vetmic.2013.10.011","article-title":"Effects of different polymerases of avian influenza viruses on the growth and pathogenicity of A\/Puerto Rico\/8\/1934 (H1N1)-derived reassorted viruses","volume":"168","author":"Kim","year":"2014","journal-title":"Vet. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Lee, C.-Y., An, S.-H., Choi, J.-G., Lee, Y.-J., Kim, J.-H., and Kwon, H.-J. (2020). Rank orders of mammalian pathogenicity-related PB2 mutations of avian influenza A viruses. Sci. Rep., 10.","DOI":"10.1038\/s41598-020-62036-5"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"7491","DOI":"10.1038\/ncomms8491","article-title":"Identification of mammalian-adapting mutations in the polymerase complex of an avian H5N1 influenza virus","volume":"6","author":"Taft","year":"2015","journal-title":"Nat. Commun."},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"An, S.-H., Hong, S.-M., Song, J.-H., Son, S.-E., Lee, C.-Y., Choi, K.-S., and Kwon, H.-J. (2023). Engineering an Optimal Y280-Lineage H9N2 Vaccine Strain by Tuning PB2 Activity. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24108840"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Mezhenskaya, D., Isakova-Sivak, I., and Rudenko, L. (2019). M2e-based universal influenza vaccines: A historical overview and new approaches to development. J. Biomed. Sci., 26.","DOI":"10.1186\/s12929-019-0572-3"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Manenti, A., Maciola, A.K., Trombetta, C.M., Kistner, O., Casa, E., Hyseni, I., Razzano, I., Torelli, A., and Montomoli, E. (2020). Influenza anti-stalk antibodies: Development of a new method for the evaluation of the immune responses to universal vaccine. Vaccines, 8.","DOI":"10.3390\/vaccines8010043"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Gu, M., Zhao, Y., Ge, Z., Li, Y., Gao, R., Wang, X., Hu, J., Liu, X., Hu, S., and Peng, D. (2022). Effects of HA2 154 deglycosylation and NA V202I mutation on biological property of H5N6 subtype avian influenza virus. Vet. Microbiol., 266.","DOI":"10.1016\/j.vetmic.2022.109353"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"8496","DOI":"10.1128\/JVI.00880-16","article-title":"Unmasking stem-specific neutralizing epitopes by abolishing N-linked glycosylation sites of influenza virus hemagglutinin proteins for vaccine design","volume":"90","author":"Liu","year":"2016","journal-title":"J. Virol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"e2074","DOI":"10.1002\/rmv.2074","article-title":"Inactivation methods for whole influenza vaccine production","volume":"29","author":"Sabbaghi","year":"2019","journal-title":"Rev. Med. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"695","DOI":"10.1586\/erv.12.38","article-title":"Inactivated virus vaccines from chemistry to prophylaxis: Merits, risks and challenges","volume":"11","author":"Delrue","year":"2012","journal-title":"Expert Rev. Vaccines"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"994","DOI":"10.7589\/0090-3558-47.4.994","article-title":"Surveillance and Isolation of HPAI H5N1 from Wild Mandarin Ducks (Aix galericulata)","volume":"47","author":"Lee","year":"2011","journal-title":"J. Wildl. Dis."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3165","DOI":"10.1016\/S0264-410X(02)00268-2","article-title":"Eight-plasmid system for rapid generation of influenza virus vaccines","volume":"20","author":"Hoffmann","year":"2002","journal-title":"Vaccine"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Iba\u00f1ez, L.I., Roose, K., De Filette, M., Schotsaert, M., De Sloovere, J., Roels, S., Pollard, C., Schepens, B., Grooten, J., and Fiers, W. (2013). M2e-Displaying Virus-Like Particles with Associated RNA Promote T Helper 1 Type Adaptive Immunity against Influenza A. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0059081"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Tsybalova, L.M., Stepanova, L.A., Shuklina, M.A., Mardanova, E.S., Kotlyarov, R.Y., Potapchuk, M.V., Petrov, S.A., Blokhina, E.A., and Ravin, N.V. (2018). Combination of M2e peptide with stalk HA epitopes of influenza A virus enhances protective properties of recombinant vaccine. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0201429"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1038\/s41541-022-00498-6","article-title":"A chimeric thermostable M2e and H3 stalk-based universal influenza A virus vaccine","volume":"7","author":"Subbiah","year":"2022","journal-title":"npj Vaccines"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"187ra170","DOI":"10.1126\/scitranslmed.3005996","article-title":"Glycosylations in the Globular Head of the Hemagglutinin Protein Modulate the Virulence and Antigenic Properties of the H1N1 Influenza Viruses","volume":"5","author":"Medina","year":"2013","journal-title":"Sci. Transl. Med."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2275","DOI":"10.1007\/s007050170002","article-title":"Universal primer set for the full-length amplification of all influenza A viruses","volume":"146","author":"Hoffmann","year":"2001","journal-title":"Arch. Virol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"714","DOI":"10.1021\/es60130a004","article-title":"Trimmed Spearman-Karber method for estimating median lethal concentrations in toxicity bioassays","volume":"11","author":"Hamilton","year":"1977","journal-title":"Environ. Sci. Technol."},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Spackman, E. (2020). Serum Virus Neutralization Assay for Detection and Quantitation of Serum Neutralizing Antibodies to Influenza A Virus in Swine. Animal Influenza Virus: Methods and Protocols, Springer. Methods in Molecular Biology.","DOI":"10.1007\/978-1-0716-0346-8"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"6154","DOI":"10.1016\/j.vaccine.2019.08.074","article-title":"Bioengineering a highly productive vaccine strain in embryonated chicken eggs and mammals from a non-pathogenic clade 2\u00b7 3\u00b7 4\u00b7 4 H5N8 strain","volume":"37","author":"An","year":"2019","journal-title":"Vaccine"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"793","DOI":"10.1038\/35021239","article-title":"Vaccines against intracellular infections requiring cellular immunity","volume":"406","author":"Seder","year":"2000","journal-title":"Nature"},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"An, S.-H., Lee, C.-Y., Hong, S.-M., Song, C.-S., Kim, J.-H., and Kwon, H.-J. (2019). Novel mutations evading avian immunity around the receptor binding site of the clade 2.3. 2.1 c hemagglutinin gene reduce viral thermostability and mammalian pathogenicity. Viruses, 11.","DOI":"10.20944\/preprints201909.0323.v1"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Siegers, J.Y., Xie, R., Byrne, A.M., Edwards, K.M., Hu, S., Yann, S., Sin, S., Tok, S., Chea, K., and Horm, S.V. (2024). Emergence of a Novel Reassortant Clade 2.3. 2.1 c Avian Influenza A\/H5N1 Virus Associated with Human Cases in Cambodia. medRxiv.","DOI":"10.1101\/2024.11.04.24313747"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"1793","DOI":"10.1080\/22221751.2020.1797542","article-title":"Evolution and extensive reassortment of H5 influenza viruses isolated from wild birds in China over the past decade","volume":"9","author":"Cui","year":"2020","journal-title":"Emerg Microbes Infect."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1050","DOI":"10.1016\/S0006-3495(02)75464-7","article-title":"Protonation and stability of the globular domain of influenza virus hemagglutinin","volume":"82","author":"Huang","year":"2002","journal-title":"Biophys. J."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Klenow, L., Elfageih, R., Gao, J., Wan, H., Withers, S.G., de Gier, J.-W., and Daniels, R. (2023). Influenza virus and pneumococcal neuraminidases enhance catalysis by similar yet distinct sialic acid\u2013binding strategies. J. Biol. Chem., 299.","DOI":"10.1016\/j.jbc.2023.102891"},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Lagache, T., Sieben, C., Meyer, T., Herrmann, A., and Holcman, D. (2017). Stochastic model of acidification, activation of hemagglutinin and escape of influenza viruses from an endosome. Front. Phys., 5.","DOI":"10.3389\/fphy.2017.00025"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"759","DOI":"10.1128\/jvi.67.2.759-764.1993","article-title":"Biologic importance of neuraminidase stalk length in influenza A virus","volume":"67","author":"Castrucci","year":"1993","journal-title":"J. Virol."},{"key":"ref_47","unstructured":"Kwon, H.-J., Choi, J.-G., Lee, Y.-J., and Kim, J.-H. (2014). A Method of preparing HA2 common epitope of influenza virus with improved immunogenicity. (KR101415836B1), KR Patent."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"e00373-22","DOI":"10.1128\/jvi.00373-22","article-title":"Hydrophobic residues at the intracellular domain of the M2 protein play an important role in budding and membrane integrity of influenza virus","volume":"96","author":"Bao","year":"2022","journal-title":"J. Virol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"5478","DOI":"10.1016\/j.bpj.2021.11.023","article-title":"Influenza A M2 recruits M1 to the plasma membrane: A fluorescence fluctuation microscopy study","volume":"120","author":"Petrich","year":"2021","journal-title":"Biophys. J."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.vetmic.2018.11.031","article-title":"Generation of highly productive and mammalian nonpathogenic recombinant H9N2 avian influenza viruses by optimization of 3\u2019end promoter and NS genome","volume":"228","author":"An","year":"2019","journal-title":"Vet. Microbiol."},{"key":"ref_51","doi-asserted-by":"crossref","unstructured":"An, S.-H., Son, S.-E., Song, J.-H., Hong, S.-M., Lee, C.-Y., Lee, N.-H., Jeong, Y.-J., Choi, J.-G., Lee, Y.-J., and Kang, H.-M. (2022). Selection of an Optimal Recombinant Egyptian H9N2 Avian Influenza Vaccine Strain for Poultry with High Antigenicity and Safety. Vaccines, 10.","DOI":"10.3390\/vaccines10020162"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Kim, J.Y., Park, S.Y., Jin, J.S., Kim, D., Park, J.-H., Park, S.H., and Ko, Y.-J. (2023). Efficacy of binary ethylenimine in the inactivation of foot-and-mouth disease virus for vaccine production in South Korea. Pathogens, 12.","DOI":"10.3390\/pathogens12060760"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1007\/BF01315592","article-title":"Binary ethylenimine as an inactivant for foot-and-mouth disease virus and its application for vaccine production","volume":"47","author":"Bahnemann","year":"1975","journal-title":"Arch. Virol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"e00840","DOI":"10.1128\/JVI.00840-19","article-title":"Extending the Stalk Enhances Immunogenicity of the Influenza Virus Neuraminidase","volume":"93","author":"Broecker","year":"2019","journal-title":"J. Virol."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1080\/0307945021000070714","article-title":"Development of a DIVA (Differentiating Infected from Vaccinated Animals) strategy using a vaccine containing a heterologous neuraminidase for the control of avian influenza","volume":"32","author":"Capua","year":"2003","journal-title":"Avian Pathol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1080\/03079450600598079","article-title":"Development and validation of an anti-N3 indirect immunofluorescent antibody test to be used as a companion diagnostic test in the framework of a \u201cDIVA\u201d vaccination strategy for avian influenza infections in poultry","volume":"35","author":"Cattoli","year":"2006","journal-title":"Avian Pathol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"1022","DOI":"10.4049\/jimmunol.0902147","article-title":"Universal vaccine based on ectodomain of matrix protein 2 of influenza A: Fc receptors and alveolar macrophages mediate protection","volume":"186","author":"Descamps","year":"2011","journal-title":"J. Immunol."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"3700","DOI":"10.1128\/JVI.02576-14","article-title":"Structure of the extracellular domain of matrix protein 2 of influenza A virus in complex with a protective monoclonal antibody","volume":"89","author":"Cho","year":"2015","journal-title":"J. Virol."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"107","DOI":"10.4049\/jimmunol.2300343","article-title":"Both Humoral and Cellular Immunity Limit the Ability of Live Attenuated Influenza Vaccines to Promote T Cell Responses","volume":"212","author":"Lobby","year":"2024","journal-title":"J. Immunol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"314","DOI":"10.1038\/s41586-021-04356-8","article-title":"Broadly neutralizing antibodies target a haemagglutinin anchor epitope","volume":"602","author":"Guthmiller","year":"2022","journal-title":"Nature"},{"key":"ref_61","first-page":"10","article-title":"Hierarchical and redundant roles of activating Fc\u03b3Rs in protection against influenza disease by M2e-specific IgG1 and IgG2a antibodies","volume":"91","author":"Ehrhardt","year":"2017","journal-title":"J. Virol."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/1\/22\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:56:51Z","timestamp":1760115411000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/1\/22"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,12,30]]},"references-count":61,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,1]]}},"alternative-id":["vaccines13010022"],"URL":"https:\/\/doi.org\/10.3390\/vaccines13010022","relation":{},"ISSN":["2076-393X"],"issn-type":[{"type":"electronic","value":"2076-393X"}],"subject":[],"published":{"date-parts":[[2024,12,30]]}}}