{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,27]],"date-time":"2026-02-27T23:14:33Z","timestamp":1772234073883,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T00:00:00Z","timestamp":1763078400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Patronus Biotech Co., Ltd."}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background: Seasonal influenza vaccines must be reformulated annually due to the high genetic variability and antigenic drift of circulating influenza viruses. The annual update, guided by World Health Organization (WHO) recommendations, results in significant challenges, including compressed production time periods, elevated manufacturing costs, and global distribution pressures. Moreover, mismatches between vaccine strains and circulating viruses can severely reduce protective efficacy, underscoring the urgent need for broadly protective and long-lasting influenza vaccines. Methods: In this study, we developed an adjuvanted trivalent recombinant influenza virus-like particle vaccine (a-RIV) using the baculovirus\u2013insect cell expression system and formulated it with an AS01-like adjuvant. The vaccine comprises full-length hemagglutinin (HA) proteins from WHO-recommended seasonal influenza strains: A\/H1N1 (AH1), A\/H3N2 (AH3), and B\/Victoria (B\/vic) lineages. The purified HA proteins were subsequently formulated with a liposomal adjuvant to enhance the immunogenicity. Results: In mouse immunization studies, the a-RIV vaccine elicited significantly stronger humoral and cellular immune responses than the licensed recombinant vaccine Flublok and the conventional inactivated influenza vaccine (IIV). High levels of functional anti-HA antibodies and antigen-specific T cell responses persisted for at least six months post-vaccination. Moreover, a-RIV induced broadly reactive antibodies capable of cross-binding to heterologous AH1 and AH3 influenza strains. Conclusions: Our data demonstrate that the a-RIV elicits enhanced, durable, and broadly cross-reactive immune responses against multiple influenza subtypes. These findings support the potential of adjuvanted recombinant HA-based vaccine as a promising candidate for the development of next-generation influenza vaccine.<\/jats:p>","DOI":"10.3390\/vaccines13111162","type":"journal-article","created":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T11:49:43Z","timestamp":1763120983000},"page":"1162","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Adjuvanted Recombinant Hemagglutinin Vaccine Provides Durable and Broad-Spectrum Immunogenicity in Mice"],"prefix":"10.3390","volume":"13","author":[{"given":"Rui","family":"Yu","sequence":"first","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5724-2110","authenticated-orcid":false,"given":"Yan","family":"Guo","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Senyan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanbao","family":"Ai","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rui","family":"Wei","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan","family":"Li","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hang","family":"Chen","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Shuyun","family":"Liu","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Caixia","family":"Zhang","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanfeng","family":"Yao","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meng","family":"Lv","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yingying","family":"Li","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yulin","family":"Chen","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peng","family":"Zhou","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siting","family":"Tu","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Meijuan","family":"Fu","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongshun","family":"Su","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Lin","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min","family":"Yang","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yanbin","family":"Ding","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siyu","family":"Tian","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cai","family":"Jing","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hang","family":"Chen","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Ma","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunping","family":"Deng","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yu","family":"Zhou","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuanyuan","family":"Li","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jing","family":"Jin","sequence":"additional","affiliation":[{"name":"Patronus Biotech Co., Ltd., Guangzhou 510005, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,11,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S2213-2600(18)30496-X","article-title":"Mortality, morbidity, and hospitalisations due to influenza lower respiratory tract infections, 2017: An analysis for the Global Burden of Disease Study 2017","volume":"7","author":"Collaborators","year":"2019","journal-title":"Lancet Respir. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"A6","DOI":"10.1016\/j.vaccine.2020.09.048","article-title":"The disease burden of influenza beyond respiratory illness","volume":"39","author":"Macias","year":"2021","journal-title":"Vaccine"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Carascal, M.B., Pavon, R.D.N., and Rivera, W.L. (2022). Rivera, Recent Progress in Recombinant Influenza Vaccine Development Toward Heterosubtypic Immune Response. Front. Immunol., 13.","DOI":"10.3389\/fimmu.2022.878943"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1021\/acsphotonics.1c01154","article-title":"Nanoscale Mapping of Recombinant Viral Proteins: From Cells to Virus-Like Particles","volume":"9","author":"Delcanale","year":"2022","journal-title":"ACS Photonics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1111\/j.1750-2659.2008.00053.x","article-title":"FluBlok, a recombinant hemagglutinin influenza vaccine","volume":"2","author":"Cox","year":"2008","journal-title":"Influ. Other Respir. Viruses"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"813","DOI":"10.1111\/irv.12871","article-title":"Effectiveness of the MF59-adjuvanted trivalent or quadrivalent seasonal influenza vaccine among adults 65 years of age or older, a systematic review and meta-analysis","volume":"15","author":"Coleman","year":"2021","journal-title":"Influ. Other Respir. Viruses"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"513","DOI":"10.1016\/j.vaccine.2016.12.011","article-title":"Effectiveness of MF59-adjuvanted seasonal influenza vaccine in the elderly: A systematic review and meta-analysis","volume":"35","author":"Domnich","year":"2017","journal-title":"Vaccine"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Wilkins, A.L., Kazmin, D., Napolitani, G., Clutterbuck, E.A., Pulendran, B., Siegrist, C.-A., and Pollard, A.J. (2017). AS03- and MF59-Adjuvanted Influenza Vaccines in Children. Front. Immunol., 8.","DOI":"10.3389\/fimmu.2017.01760"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1013","DOI":"10.1007\/s40265-024-02083-8","article-title":"Influenza Vaccine Effectiveness and Progress Towards a Universal Influenza Vaccine","volume":"84","author":"Cowling","year":"2024","journal-title":"Drugs"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1564","DOI":"10.1093\/cid\/ciw635","article-title":"2014\u20132015 Influenza Vaccine Effectiveness in the United States by Vaccine Type","volume":"63","author":"Zimmerman","year":"2016","journal-title":"Clin. Infect. Dis."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1898","DOI":"10.1016\/j.chom.2023.09.012","article-title":"An atlas of continuous adaptive evolution in endemic human viruses","volume":"31","author":"Kistler","year":"2023","journal-title":"Cell Host Microbe"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"a038489","DOI":"10.1101\/cshperspect.a038489","article-title":"The Ecology and Evolution of Influenza Viruses","volume":"10","author":"Wille","year":"2020","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Nuwarda, R.F., Alharbi, A.A., and Kayser, V. (2021). An Overview of Influenza Viruses and Vaccines. Vaccines, 9.","DOI":"10.3390\/vaccines9091032"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"a038638","DOI":"10.1101\/cshperspect.a038638","article-title":"Hemagglutinin Structure and Activities","volume":"11","author":"Gamblin","year":"2021","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"132","DOI":"10.1038\/s41541-023-00728-5","article-title":"A mosaic influenza virus-like particles vaccine provides broad humoral and cellular immune responses against influenza A viruses","volume":"8","author":"Liu","year":"2023","journal-title":"npj Vaccines"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1038\/nature18010","article-title":"Design of a hyperstable 60-subunit protein icosahedron","volume":"535","author":"Hsia","year":"2016","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8855","DOI":"10.1021\/acsnano.8b02805","article-title":"Engineering a Rugged Nanoscaffold To Enhance Plug-and-Display Vaccination","volume":"12","author":"Bruun","year":"2018","journal-title":"ACS Nano"},{"key":"ref_18","first-page":"56448","article-title":"Measuring Influenza Neutralizing Antibody Responses to A(H3N2) Viruses in Human Sera by Microneutralization Assays Using MDCK-SIAT1 Cells","volume":"129","author":"Gross","year":"2017","journal-title":"J. Vis. Exp."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"4975","DOI":"10.1016\/j.vaccine.2015.07.026","article-title":"Recombinant H7 hemagglutinin forms subviral particles that protect mice and ferrets from challenge with H7N9 influenza virus","volume":"33","author":"Pushko","year":"2015","journal-title":"Vaccine"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"27","DOI":"10.4103\/0976-0105.177703","article-title":"A simple practice guide for dose conversion between animals and human","volume":"7","author":"Nair","year":"2016","journal-title":"J. Basic Clin. Pharm."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1016\/j.jcv.2019.08.009","article-title":"Influenza virus-specific CD4+ and CD8+ T cell-mediated immunity induced by infection and vaccination","volume":"119","author":"Jansen","year":"2019","journal-title":"J. Clin. Virol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1038\/nm.3350","article-title":"Cellular immune correlates of protection against symptomatic pandemic influenza","volume":"19","author":"Sridhar","year":"2013","journal-title":"Nat. Med."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Langat, P., Raghwani, J., Dudas, G., Bowden, T.A., Edwards, S., Gall, A., Bedford, T., Rambaut, A., Daniels, R.S., and Russell, C.A. (2017). Genome-wide evolutionary dynamics of influenza B viruses on a global scale. PLOS Pathog., 13.","DOI":"10.1371\/journal.ppat.1006749"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1038\/nature06945","article-title":"The genomic and epidemiological dynamics of human influenza A virus","volume":"453","author":"Rambaut","year":"2008","journal-title":"Nature"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1128\/CVI.00085-16","article-title":"Characterization of Influenza Vaccine Hemagglutinin Complexes by Cryo-Electron Microscopy and Image Analyses Reveals Structural Polymorphisms","volume":"23","author":"McCraw","year":"2016","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1038\/s41467-023-37162-z","article-title":"Commercial influenza vaccines vary in HA-complex structure and in induction of cross-reactive HA antibodies","volume":"14","author":"Myers","year":"2023","journal-title":"Nat. Commun."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1038\/s41541-021-00403-7","article-title":"Unique features of a recombinant haemagglutinin influenza vaccine that influence vaccine performance","volume":"6","author":"Arunachalam","year":"2021","journal-title":"npj Vaccines"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"159","DOI":"10.3947\/ic.2013.45.2.159","article-title":"Fluad\u00ae-MF59\u00ae-Adjuvanted Influenza Vaccine in Older Adults","volume":"45","author":"Tsai","year":"2013","journal-title":"Infect. Chemother."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"740","DOI":"10.1038\/sj.clpt.6100402","article-title":"MF59 is a safe and potent vaccine adjuvant for flu vaccines in humans: What did we learn during its development?","volume":"82","author":"Wack","year":"2007","journal-title":"Clin. Pharmacol. Ther."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3363","DOI":"10.1016\/j.vaccine.2013.05.007","article-title":"The adjuvant effect of MF59 is due to the oil-in-water emulsion formulation, none of the individual components induce a comparable adjuvant effect","volume":"31","author":"Calabro","year":"2013","journal-title":"Vaccine"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"2189885","DOI":"10.1080\/21645515.2023.2189885","article-title":"The Matrix-M adjuvant: A critical component of vaccines for the 21(st) century","volume":"19","author":"Stertman","year":"2023","journal-title":"Hum. Vaccin. Immunother."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1920","DOI":"10.4049\/jimmunol.1400948","article-title":"Enhancement of adaptive immunity by the human vaccine adjuvant AS01 depends on activated dendritic cells","volume":"193","author":"Didierlaurent","year":"2014","journal-title":"J. Immunol."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Bosteels, C., Fierens, K., De Prijck, S., Van Moorleghem, J., Vanheerswynghels, M., De Wolf, C., Chalon, A., Collignon, C., Hammad, H., and Didierlaurent, A.M. (2020). CCR2- and Flt3-Dependent Inflammatory Conventional Type 2 Dendritic Cells Are Necessary for the Induction of Adaptive Immunity by the Human Vaccine Adjuvant System AS01. Front. Immunol., 11.","DOI":"10.3389\/fimmu.2020.606805"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"e4278","DOI":"10.1093\/cid\/ciaa1673","article-title":"Induction of Cross-Reactive Hemagglutination Inhibiting Antibody and Polyfunctional CD4+ T-Cell Responses by a Recombinant Matrix-M\u2013Adjuvanted Hemagglutinin Nanoparticle Influenza Vaccine","volume":"73","author":"Shinde","year":"2021","journal-title":"Clin. Infect. Dis."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"715","DOI":"10.1080\/14760584.2024.2382725","article-title":"Adjuvant system AS01: From mode of action to effective vaccines","volume":"23","author":"Roman","year":"2024","journal-title":"Expert Rev. Vaccines"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"RTS,S Clinical Trials Partnership, Agnandji, S.T., Lell, B., Soulanoudjingar, S.S., Fernandes, J.F., Abossolo, B.P., Conzelmann, C., Methogo, B.G., Doucka, Y., and Flamen, A. (2011). First Results of Phase 3 Trial of RTS,S\/AS01 Malaria Vaccine in African Children. N. Engl. J. Med., 365, 1863\u20131875.","DOI":"10.1056\/NEJMoa1102287"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Silva, G.S.D., Borges, S.G., Pozzebon, B.B., and Souza, A.P.D. (2024). Immune Responses to Respiratory Syncytial Virus Vaccines: Advances and Challenges. Microorganisms, 12.","DOI":"10.3390\/microorganisms12112305"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1177\/0018578718767103","article-title":"Zoster Vaccine Recombinant, Adjuvanted","volume":"53","author":"Stoker","year":"2018","journal-title":"Hosp. Pharm."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"405","DOI":"10.4161\/hv.22908","article-title":"Correlates of protection to influenza virus, where do we go from here?","volume":"9","author":"Cox","year":"2013","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1126\/science.1083317","article-title":"Defective CD8 T cell memory following acute infection without CD4 T cell help","volume":"300","author":"Sun","year":"2003","journal-title":"Science"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"274","DOI":"10.1038\/nm.2612","article-title":"Preexisting influenza-specific CD4+ T cells correlate with disease protection against influenza challenge in humans","volume":"18","author":"Wilkinson","year":"2012","journal-title":"Nat. Med."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Duan, S., and Thomas, P.G. (2016). Balancing Immune Protection and Immune Pathology by CD8+ T-Cell Responses to Influenza Infection. Front. Immunol., 7.","DOI":"10.3389\/fimmu.2016.00025"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.virusres.2011.09.022","article-title":"Immune responses to influenza virus infection","volume":"162","author":"Kreijtz","year":"2011","journal-title":"Virus Res."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/11\/1162\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,14]],"date-time":"2025-11-14T12:14:54Z","timestamp":1763122494000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/11\/1162"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,14]]},"references-count":43,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2025,11]]}},"alternative-id":["vaccines13111162"],"URL":"https:\/\/doi.org\/10.3390\/vaccines13111162","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,14]]}}}