{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T01:21:12Z","timestamp":1760059272815,"version":"build-2065373602"},"reference-count":31,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2025,5,31]],"date-time":"2025-05-31T00:00:00Z","timestamp":1748649600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the National Research Foundation of Korea (NRF)","award":["NRF2018R1A6A1A03025124"],"award-info":[{"award-number":["NRF2018R1A6A1A03025124"]}]},{"name":"Kyung Hee University","award":["NRF2018R1A6A1A03025124"],"award-info":[{"award-number":["NRF2018R1A6A1A03025124"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background: Self-administered orally dissolving films (ODFs) encapsulating inactivated influenza vaccines represent an effective strategy for stimulating mucosal immunity. While this vaccination method offers several advantages over conventional influenza vaccines, a comparative efficacy study remains lacking. Methods: Female BALB\/c mice were immunized with inactivated A\/PR\/8\/34 (H1N1) either via orogastric inoculation or through the oral mucosal delivery using pullulan and trehalose-based ODF vaccines. Each group received equivalent antigen doses across three immunizations. Humoral responses and antibody functionality were assessed using sera collected post-immunization. After lethal viral challenge, other immunological and virological parameters were determined in corresponding tissues. Body weight and survival were monitored over a 14-day period after challenge. Results: ODF vaccination elicited significantly higher virus-specific IgA levels, HAI titers, and neutralizing antibody activity than oral gavage. After the viral challenge, ODF-immunized mice exhibited stronger IgG and IgA responses in respiratory tissues, increased antibody-secreting cells in lungs and spleen, and elevated germinal center B cells and CD8+ T cell responses. Both vaccination methods reduced lung pro-inflammatory cytokines and provided full protection against lethal challenge; however, the ODF group showed lower cytokine levels, better weight maintenance, and reduced viral loads. Conclusions: ODF vaccination elicits more robust systemic and mucosal immune responses than oral vaccination and may serve as a promising alternative method of influenza vaccine delivery.<\/jats:p>","DOI":"10.3390\/vaccines13060600","type":"journal-article","created":{"date-parts":[[2025,6,2]],"date-time":"2025-06-02T11:40:32Z","timestamp":1748864432000},"page":"600","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Orally Dissolving Film-Based Influenza Vaccines Confer Superior Protection Compared to the Oral Administration of Inactivated Influenza Virus"],"prefix":"10.3390","volume":"13","author":[{"given":"Keon-Woong","family":"Yoon","sequence":"first","affiliation":[{"name":"Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea"},{"name":"Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06510, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jie","family":"Mao","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea"},{"name":"Medical Research Center for Bioreaction to Reactive Oxygen Species and Biomedical Science Institute, Core Research Institute (CRI), Kyung Hee University, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gi-Deok","family":"Eom","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea"},{"name":"Medical Research Center for Bioreaction to Reactive Oxygen Species and Biomedical Science Institute, Core Research Institute (CRI), Kyung Hee University, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Su In","family":"Heo","sequence":"additional","affiliation":[{"name":"Department of Biomedical Science, Graduate School, Kyung Hee University, Seoul 02447, Republic of Korea"},{"name":"Medical Research Center for Bioreaction to Reactive Oxygen Species and Biomedical Science Institute, Core Research Institute (CRI), Kyung Hee University, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ki Back","family":"Chu","sequence":"additional","affiliation":[{"name":"Department of Parasitology, College of Medicine, Inje University, Busan 47392, Republic of Korea"},{"name":"Department of Infectious Disease and Malaria, Paik Institute of Clinical Research, Inje University, Busan 47392, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mi Suk","family":"Lee","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, Department of Internal Medicine, College of Medicine, Kyung Hee University, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0419-9339","authenticated-orcid":false,"given":"Fu-Shi","family":"Quan","sequence":"additional","affiliation":[{"name":"Medical Research Center for Bioreaction to Reactive Oxygen Species and Biomedical Science Institute, Core Research Institute (CRI), Kyung Hee University, Seoul 02447, Republic of Korea"},{"name":"Department of Medical Zoology, School of Medicine, Kyung Hee University, Seoul 02447, Republic of Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,31]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1007\/s40121-024-01082-2","article-title":"Respiratory Virus Vaccines: Pathways to Recommendations and Enhanced Coverage for At-Risk Populations","volume":"14","author":"Maggi","year":"2025","journal-title":"Infect. Dis. Ther."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1","DOI":"10.15585\/mmwr.rr7202a1","article-title":"Prevention and control of seasonal influenza with vaccines: Recommendations of the Advisory Committee on Immunization Practices\u2014United States, 2023\u201324 influenza season","volume":"72","author":"Grohskopf","year":"2023","journal-title":"MMWR Recomm. Rep."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"619","DOI":"10.1007\/s11739-023-03456-9","article-title":"Influenza vaccination for elderly, vulnerable and high-risk subjects: A narrative review and expert opinion","volume":"19","author":"Consoli","year":"2024","journal-title":"Intern. Emerg. Med."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1038\/nm0103-99","article-title":"Can needle-free administration of vaccines become the norm in global immunization?","volume":"9","author":"Levine","year":"2003","journal-title":"Nat. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1080\/17425247.2020.1731468","article-title":"Non-invasive mucosal vaccine delivery: Advantages, challenges and the future","volume":"17","author":"Skwarczynski","year":"2020","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"180","DOI":"10.1016\/j.vaccine.2011.11.028","article-title":"Oral vaccination with inactivated influenza vaccine induces cross-protective immunity","volume":"30","author":"Quan","year":"2012","journal-title":"Vaccine"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Tian, Y., Bhide, Y.C., Woerdenbag, H.J., Huckriede, A.L., Frijlink, H.W., Hinrichs, W.L., and Visser, J.C. (2020). Development of an orodispersible film containing stabilized influenza vaccine. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12030245"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.jconrel.2015.03.012","article-title":"Oral films: Current status and future perspectives II\u2014Intellectual property, technologies and market needs","volume":"206","author":"Borges","year":"2015","journal-title":"J. Control. Release"},{"key":"ref_9","first-page":"6391","article-title":"Challenges in formulation development of fast dissolving oral films","volume":"3","author":"Jadhav","year":"2013","journal-title":"J. Pharm Res"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"105979","DOI":"10.1016\/j.antiviral.2024.105979","article-title":"Orally dissolving film as a potential vaccine delivery carrier to prevent influenza virus infection","volume":"230","author":"Yoon","year":"2024","journal-title":"Antivir. Res."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"124945","DOI":"10.1016\/j.ijpharm.2024.124945","article-title":"Dose sparing enabled by immunization with influenza vaccine using orally dissolving film","volume":"667","author":"Yoon","year":"2024","journal-title":"Int. J. Pharm."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"193","DOI":"10.3347\/PHD.24017","article-title":"Virus-like particles expressing microneme-associated antigen of Plasmodium berghei confer better protection than those expressing apical membrane antigen 1","volume":"62","author":"Kim","year":"2024","journal-title":"Parasites Hosts Dis."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"106374","DOI":"10.1016\/j.ejps.2023.106374","article-title":"Advances in formulation and manufacturing strategies for the delivery of therapeutic proteins and peptides in orally disintegrating dosage forms","volume":"182","author":"Bashir","year":"2023","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"120759","DOI":"10.1016\/j.ijpharm.2021.120759","article-title":"Recent advances of oral film as platform for drug delivery","volume":"604","author":"He","year":"2021","journal-title":"Int. J. Pharm."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1016\/j.ejpb.2018.09.016","article-title":"Orodispersible films based on blends of trehalose and pullulan for protein delivery","volume":"133","author":"Tian","year":"2018","journal-title":"Eur. J. Pharm. Biopharm."},{"key":"ref_16","unstructured":"Tian, Y., Visser, J.C., Frijlink, H., and Woerdenbag, H. (2018, January 19\u201322). Pullulan and trehalose based films for protein delivery in the oral cavity. Proceedings of the 11th World Meeting on Pharmaceutics, Biopharmaceutics and Pharmaceutical Technology\u2014Palacio de Exposiciones y Congressos Granada, Granada, Spain."},{"key":"ref_17","first-page":"9","article-title":"Orally fast dissolving films: Innovations in formulation and technology","volume":"9","author":"Bhyan","year":"2011","journal-title":"Int. J. Pharm. Sci. Rev. Res."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Chun, M.-H., Kim, J.Y., Park, E.-S., and Choi, D.H. (2021). Development of a robust control strategy for fixed-dose combination bilayer tablets with integrated quality by design, statistical, and process analytical technology approach. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13091443"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"656","DOI":"10.1038\/nrmicro2384","article-title":"The role of secretory antibodies in infection immunity","volume":"8","author":"Strugnell","year":"2010","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"236","DOI":"10.7883\/yoken.JJID.2004.236","article-title":"Defense mechanisms against influenza virus infection in the respiratory tract mucosa","volume":"57","author":"Tamura","year":"2004","journal-title":"Jpn. J. Infect. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"322","DOI":"10.1016\/j.ymeth.2009.04.015","article-title":"Translational modifications to improve vaccine efficacy in an oral influenza vaccine","volume":"49","author":"Bennett","year":"2009","journal-title":"Methods"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"323","DOI":"10.1517\/17425247.4.4.323","article-title":"Oral vaccination: Where we are?","volume":"4","author":"Silin","year":"2007","journal-title":"Expert Opin. Drug Deliv."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3456","DOI":"10.1016\/j.apsb.2022.07.002","article-title":"Mucosal vaccine delivery: A focus on the breakthrough of specific barriers","volume":"12","author":"Huang","year":"2022","journal-title":"Acta Pharm. Sin. B"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"S45","DOI":"10.1038\/nm1213","article-title":"Mucosal immunity and vaccines","volume":"11","author":"Holmgren","year":"2005","journal-title":"Nat. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1796","DOI":"10.4049\/jimmunol.168.4.1796","article-title":"Nasal-associated lymphoid tissue is a mucosal inductive site for virus-specific humoral and cellular immune responses","volume":"168","author":"Zuercher","year":"2002","journal-title":"J. Immunol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Czerkinsky, C., and Holmgren, J. (2010). Mucosal delivery routes for optimal immunization: Targeting immunity to the right tissues. Mucosal Vaccines: Modern Concepts, Strategies, and Challenges, Springer.","DOI":"10.1007\/82_2010_112"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1089\/vim.2017.0025","article-title":"Adaptive B cell responses to influenza virus infection in the lung","volume":"30","author":"Takahashi","year":"2017","journal-title":"Viral Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"667","DOI":"10.1038\/nri3738","article-title":"Regional specialization within the intestinal immune system","volume":"14","author":"Mowat","year":"2014","journal-title":"Nat. Rev. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1016\/j.coviro.2013.05.001","article-title":"Role of T cell immunity in recovery from influenza virus infection","volume":"3","author":"Sun","year":"2013","journal-title":"Curr. Opin. Virol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1080\/14760584.2018.1541407","article-title":"Basics of CD8 T-cell immune responses after influenza infection and vaccination with inactivated or live attenuated influenza vaccine","volume":"17","author":"Korenkov","year":"2018","journal-title":"Expert Rev. Vaccines"},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Jia, J., Li, H., Huang, Z., Yu, J., Zheng, Y., and Cao, B. (2023). Comprehensive immune landscape of lung-resident memory CD8+ T cells after influenza infection and reinfection in a mouse model. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1184884"}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/6\/600\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T17:44:46Z","timestamp":1760031886000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/13\/6\/600"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,5,31]]},"references-count":31,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2025,6]]}},"alternative-id":["vaccines13060600"],"URL":"https:\/\/doi.org\/10.3390\/vaccines13060600","relation":{},"ISSN":["2076-393X"],"issn-type":[{"type":"electronic","value":"2076-393X"}],"subject":[],"published":{"date-parts":[[2025,5,31]]}}}