{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T21:27:24Z","timestamp":1790544444534,"version":"4.1.0"},"reference-count":41,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2022,10,9]],"date-time":"2022-10-09T00:00:00Z","timestamp":1665273600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Shanghai Science and Technology Support Program","award":["21S11909700"],"award-info":[{"award-number":["21S11909700"]}]},{"name":"Shanghai Science and Technology Support Program","award":["21S11903200"],"award-info":[{"award-number":["21S11903200"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Since some cases of human infections with H5N8 avian influenza virus have been reported and caused great concern in recent years, it is important to develop an effective vaccine for human use to prevent a potential H5N8 pandemic. In the present study, a vaccine candidate virus based on newly human-infected A\/Astrakhan\/3212\/2020 H5N8 virus was constructed by reverse genetics (RG) technology. The immunogenicity of H5N8 whole virion inactivated vaccine was evaluated by various doses of vaccine antigen formulated with squalene-based adjuvant (AddaVax), aluminum hydroxide (Al(OH)3) or without adjuvant in mice. The results showed AddaVax-adjuvanted H5N8 inactivated vaccine could stimulate the mice to produce a stronger protective immune response with higher titers of IgG antibodies, hemagglutination inhibition (HI), neuraminidase inhibition (NI) and microneutralization (MN) antibodies than vaccine formulations with Al(OH)3 adjuvant or without adjuvant, and achieve a dose-sparing effect. Moreover, the AddaVax-adjuvanted formulation also exhibited potent cross-reactive response in HI antibodies against different clades of H5 viruses. A significant correlation and a curve fitting among HI, NI and MN were found by the correlation analysis to predict the protective effect of the vaccine. With these findings, our study demonstrates that AddaVax adjuvant can enhance the immunogenicity of H5N8 inactivated vaccine remarkably, and proposes an effective strategy for dealing with a potential H5N8 virus pandemic.<\/jats:p>","DOI":"10.3390\/vaccines10101683","type":"journal-article","created":{"date-parts":[[2022,10,11]],"date-time":"2022-10-11T06:13:27Z","timestamp":1665468807000},"page":"1683","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":25,"title":["AddaVax-Adjuvanted H5N8 Inactivated Vaccine Induces Robust Humoral Immune Response against Different Clades of H5 Viruses"],"prefix":"10.3390","volume":"10","author":[{"given":"Feixia","family":"Gao","sequence":"first","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xueying","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yudong","family":"Dang","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Duan","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenting","family":"Xu","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shilei","family":"Wang","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Luo","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiuling","family":"Li","sequence":"additional","affiliation":[{"name":"Shanghai Institute of Biological Products, Shanghai 200052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,10,9]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"104885","DOI":"10.1016\/j.meegid.2021.104885","article-title":"Evolutionary history of H5 highly pathogenic avian influenza viruses (clade 2.3.4.4c) circulating in Taiwan during 2015\u20132018","volume":"92","author":"Huang","year":"2021","journal-title":"Infect. Genet. Evol."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Evolution Working Group (2008). Toward a unified nomenclature system for highly pathogenic avian influenza virus (H5N1). Emerg. Infect. Dis., 14, e1.","DOI":"10.3201\/eid1407.071681"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"2515135518821625","DOI":"10.1177\/2515135518821625","article-title":"A review of H5Nx avian influenza viruses","volume":"7","author":"Nunez","year":"2019","journal-title":"Ther. Adv. Vaccines Immunother."},{"key":"ref_4","unstructured":"WHO (2022). Cumulative Number of Confirmed Human Cases for Avian Influenza A(H5N1) Reported to WHO, 2003\u20132021, WHO."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"784","DOI":"10.1126\/science.abg6302","article-title":"Emerging H5N8 avian influenza viruses","volume":"372","author":"Shi","year":"2021","journal-title":"Science"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"484","DOI":"10.1016\/j.vaccine.2018.11.064","article-title":"Preclinical evaluation of the efficacy of an H5N8 vaccine candidate (IDCDC-RG43A) in mouse and ferret models for pandemic preparedness","volume":"37","author":"Jeong","year":"2018","journal-title":"Vaccine"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1007\/s00705-021-05284-8","article-title":"H5 cleavage-site peptide vaccine protects chickens from lethal infection by highly pathogenic H5 avian influenza viruses","volume":"167","author":"Jang","year":"2022","journal-title":"Arch. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3200","DOI":"10.1111\/tbed.14279","article-title":"Pathological analysis and genetic characterization of the first outbreak H5N8 subtype avian influenza virus isolated from wild swan in Shandong, China","volume":"68","author":"Liu","year":"2021","journal-title":"Transbound. Emerg. Dis."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Mostafa, A., Abdelwhab, E.M., Mettenleiter, T.C., and Pleschka, S. (2018). Zoonotic Potential of Influenza A Viruses: A Comprehensive Overview. Viruses, 10.","DOI":"10.3390\/v10090497"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Moatasim, Y., Kandeil, A., Mostafa, A., Kutkat, O., Sayes, M., El Taweel, A., AlKhazindar, M., AbdElSalam, E., El-Shesheny, R., and Kayali, G. (2021). Impact of Individual Viral Gene Segments from Influenza A\/H5N8 Virus on the Protective Efficacy of Inactivated Subtype-Specific Influenza Vaccine. Pathogens, 10.","DOI":"10.3390\/pathogens10030368"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1597","DOI":"10.1038\/nm.3409","article-title":"Key roles of adjuvants in modern vaccines","volume":"19","author":"Reed","year":"2013","journal-title":"Nat. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1016\/j.smim.2018.05.001","article-title":"Correlates of adjuvanticity: A review on adjuvants in licensed vaccines","volume":"39","author":"Giuseppe","year":"2018","journal-title":"Semin. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1038\/s41541-020-0187-4","article-title":"H7N9 influenza split vaccine with SWE oil-in-water adjuvant greatly enhances cross-reactive humoral immunity and protection against severe pneumonia in ferrets","volume":"5","author":"Spijkers","year":"2020","journal-title":"npj Vaccines"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"15ra5","DOI":"10.1126\/scitranslmed.3000624","article-title":"Vaccines with MF59 Adjuvant Expand the Antibody Repertoire to Target Protective Sites of Pandemic Avian H5N1 Influenza Virus","volume":"2","author":"Khurana","year":"2010","journal-title":"Sci. Transl. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1089\/mab.2018.0022","article-title":"Advancing Freund\u2019s and AddaVax Adjuvant Regimens Using CpG Oligodeoxynucleotides","volume":"37","author":"Hawksworth","year":"2018","journal-title":"Monoclon. Antib. Immunodiagn. Immunother."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Ramanunninair, M., Le, J., Onodera, S., Fulvini, A.A., Pokorny, B.A., Silverman, J., Devis, R., Arroyo, J.M., He, Y., and Boyne, A. (2013). Molecular Signature of High Yield (Growth) Influenza A Virus Reassortants Prepared as Candidate Vaccine Seeds. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0065955"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"6108","DOI":"10.1073\/pnas.100133697","article-title":"A DNA transfection system for generation of influenza A virus from eight plasmids","volume":"97","author":"Hoffmann","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_18","first-page":"670","article-title":"Development and Assessment of Two Highly Pathogenic Avian Influenza (HPAI) H5N6 Candidate Vaccine Viruses for Pandemic Preparedness","volume":"33","author":"Liu","year":"2020","journal-title":"Biomed. Environ. Sci."},{"key":"ref_19","first-page":"770","article-title":"Development and Validation of Alternative Method for Determination of Haemagglutinin Content in Influenza Vaccine","volume":"23","author":"Shao","year":"2010","journal-title":"Chin. J. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Gao, F., Yang, T., Liu, X., Xiong, F., Luo, J., Yi, Y., Fan, J., Chen, Z., and Tan, W.-S. (2020). MiRNA Targeted NP Genome of Live Attenuated Influenza Vaccines Provide Cross-Protection against a Lethal Influenza Virus Infection. Vaccines, 8.","DOI":"10.3390\/vaccines8010065"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1186\/s12929-020-00645-y","article-title":"Development of high-growth influenza H7N9 prepandemic candidate vaccine viruses in suspension MDCK cells","volume":"27","author":"Tzeng","year":"2020","journal-title":"J. Biomed. Sci."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1093\/oxfordjournals.aje.a118408","article-title":"A simple method of estimating fifty per cent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am. J. Epidemiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"78","DOI":"10.1080\/22221751.2019.1708214","article-title":"Protective efficacy of anti-neuraminidase monoclonal antibodies against H7N9 influenza virus infection","volume":"9","author":"Xiong","year":"2020","journal-title":"Emerg. Microbes Infect."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e00750-20","DOI":"10.1128\/JCM.00750-20","article-title":"Comparability of Titers of Antibodies against Seasonal Influenza Virus Strains as Determined by Hemagglutination Inhibition and Microneutralization Assays","volume":"58","author":"Heeringa","year":"2020","journal-title":"J. Clin. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"e00417","DOI":"10.1128\/mBio.00417-16","article-title":"Evaluation of Antihemagglutinin and Antineuraminidase Antibodies as Correlates of Protection in an Influenza A\/H1N1 Virus Healthy Human Challenge Model","volume":"7","author":"Memoli","year":"2016","journal-title":"mBio"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.jviromet.2014.09.003","article-title":"An optimized enzyme-linked lectin assay to measure influenza A virus neuraminidase inhibition antibody titers in human sera","volume":"210","author":"Couzens","year":"2014","journal-title":"J. Virol. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"666","DOI":"10.1093\/infdis\/jiq093","article-title":"Safety and Immunogenicity of Influenza A H5 Subunit Vaccines: Effect of Vaccine Schedule and Antigenic Variant","volume":"203","author":"Belshe","year":"2011","journal-title":"J. Infect. Dis."},{"key":"ref_28","first-page":"136","article-title":"A phase II, randomised clinical trial to demonstrate the non-inferiority of low-dose MF59\u00c2\u00ae-adjuvanted pre-pandemic A\/H5N1 influenza vaccine in adult and elderly subjects","volume":"53","author":"Czajka","year":"2012","journal-title":"J. Prev. Med. Hyg."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1080\/21645515.2014.1004026","article-title":"Advances in aluminum hydroxide-based adjuvant research and its mechanism","volume":"11","author":"He","year":"2015","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1586\/14760584.6.5.699","article-title":"MF59 is a safe and potent vaccine adjuvant that enhances protection against influenza virus infection","volume":"6","year":"2007","journal-title":"Expert Rev. Vaccines"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4341","DOI":"10.1016\/j.vaccine.2011.09.061","article-title":"The mechanism of action of MF59\u2014An innately attractive adjuvant formulation","volume":"30","author":"Ott","year":"2012","journal-title":"Vaccine"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1853","DOI":"10.1016\/j.vaccine.2018.02.065","article-title":"Engineering a stable CHO cell line for the expression of a MERS-coronavirus vaccine antigen","volume":"36","author":"Nyon","year":"2018","journal-title":"Vaccine"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"270","DOI":"10.1208\/s12249-021-02145-0","article-title":"AddaVax Formulated with PolyI:C as a Potential Adjuvant of MDCK-based Influenza Vaccine Enhances Local, Cellular, and Antibody Protective Immune Response in Mice","volume":"22","author":"Nian","year":"2021","journal-title":"AAPS PharmSciTech"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1220","DOI":"10.1038\/s41598-020-79427-3","article-title":"A chemokine-fusion vaccine targeting immature dendritic cells elicits elevated antibody responses to malaria sporozoites in infant macaques","volume":"11","author":"Luo","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"967","DOI":"10.1586\/14760584.2016.1164046","article-title":"Influenza immunology evaluation and correlates of protection: A focus on vaccines","volume":"15","author":"Trombetta","year":"2016","journal-title":"Expert Rev. Vaccines"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Versage, E., van Twuijver, E., Jansen, W., Theeuwes, A., Sawlwin, D., and Hohenboken, M. (2021). Analyses of Safety Profile and Homologous Antibody Responses to a Mammalian Cell-Based, MF59-Adjuvanted, A\/H5N1, Pandemic Influenza Vaccine across Four Phase II\/III Clinical Trials in Healthy Children, Adults, and Older Adults. Vaccines, 9.","DOI":"10.3390\/vaccines9121468"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"6930","DOI":"10.1016\/j.vaccine.2021.10.010","article-title":"Antibody responses against heterologous A\/H5N1 strains for an MF59-adjuvanted cell culture\u2013derived A\/H5N1 (aH5N1c) influenza vaccine in healthy pediatric subjects","volume":"39","author":"Chanthavanich","year":"2021","journal-title":"Vaccine"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Antigua, K.J.C., Choi, W.-S., Baek, Y.H., and Song, M.-S. (2019). The Emergence and Decennary Distribution of Clade 2.3.4.4 HPAI H5Nx. Microorganisms, 7.","DOI":"10.3390\/microorganisms7060156"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1007\/s00705-018-4062-9","article-title":"Development and comparison of two H5N8 influenza A vaccine candidate strains","volume":"164","author":"Lee","year":"2019","journal-title":"Arch. Virol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"3053","DOI":"10.1128\/JVI.02434-12","article-title":"Neuraminidase-Inhibiting Antibody Is a Correlate of Cross-Protection against Lethal H5N1 Influenza Virus in Ferrets Immunized with Seasonal Influenza Vaccine","volume":"87","author":"Rockman","year":"2013","journal-title":"J. Virol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"747774","DOI":"10.3389\/fimmu.2021.747774","article-title":"Matrix M Adjuvanted H5N1 Vaccine Elicits Broadly Neutralizing Antibodies and Neuraminidase Inhibiting Antibodies in Humans That Correlate with In Vivo Protection","volume":"12","author":"Zhou","year":"2021","journal-title":"Front. Immunol."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/10\/10\/1683\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:48:29Z","timestamp":1760143709000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/10\/10\/1683"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,10,9]]},"references-count":41,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2022,10]]}},"alternative-id":["vaccines10101683"],"URL":"https:\/\/doi.org\/10.3390\/vaccines10101683","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,10,9]]}}}