{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,30]],"date-time":"2026-10-30T13:56:08Z","timestamp":1793368568210,"version":"4.5.0"},"reference-count":38,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T00:00:00Z","timestamp":1742428800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Immunol."],"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Highly pathogenic H5Nx avian influenza (HPAI) poses a significant threat to poultry health globally, necessitating the development of effective vaccination strategies.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>This study assessed the immunogenicity and efficacy of a reverse-genetics-derived, Differentiating Infected from Vaccinated Animals (DIVA)-compatible inactivated H5N8 vaccine based on the IDCDC-RG71A strain. The vaccine was formulated with different adjuvants, including Montanide ISA 78 VG, ISA 71 R VG, GEL P PR, and mannose-conjugated chitosan nanoparticles, and administered via either the subcutaneous (SC) or intranasal (IN) route. To evaluate safety, the vaccine was tested in specific antibody negative (SAN) chickens, showing no adverse effects. Immunogenicity was assessed by measuring hemagglutination inhibition (HI) antibody titers, antigen-specific IgA and IgY levels, and CD4+ and CD8+ T cell proliferation. Vaccine efficacy was determined through a challenge study using a field isolate of H5N1.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>This showed that a single SC dose of vaccine containing ISA 78 VG or ISA 71 R VG provided the best efficacy against infection, with high survival rates, control of abnormally high temperature incidence, reduced virus shedding, and reduced lung and liver lesions. The ISA 78 VG-adjuvanted SC vaccine induced the highest HI titers and CD4+ T cell proliferation, while ISA 71 R VG and GEL P PR elicited the strongest IgY responses. In contrast, IN formulations induced IgA in the lungs and trachea however, even after two doses, failed to generate high HI titers and provided poor, if any, protection against infection. This highlights the superior efficacy of the SC over the IN route of vaccination for reducing H5N1 viral shedding.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>These results underscore the importance of both the adjuvants and delivery route to maximize HPAI vaccine efficacy. This presented system could thereby be used to develop potent and DIVA-compatible vaccines to enhance biosecurity and disease management in regions affected by endemic HPAI.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fimmu.2025.1539492","type":"journal-article","created":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T08:00:24Z","timestamp":1742457624000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":8,"title":["Evaluation of safety, immunogenicity, and efficacy of inactivated reverse-genetics-based H5N8 highly pathogenic avian influenza virus vaccine with various adjuvants via parenteral and mucosal routes in chickens"],"prefix":"10.3389","volume":"16","author":[{"given":"Kairat","family":"Tabynov","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aidana","family":"Kuanyshbek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leila","family":"Yelchibayeva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuantay","family":"Zharmambet","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zauresh","family":"Zhumadilova","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gleb","family":"Fomin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikolai","family":"Petrovsky","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olaitan C.","family":"Shekoni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gourapura J.","family":"Renukaradhya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kaissar","family":"Tabynov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2025,3,20]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","DOI":"10.1016\/j.virs.2024.04.004","article-title":"Evolutionary dynamics and comparative pathogenicity of clade 2.3.4.4b H5 subtype avian influenza viruses, China, 2021-2022","volume":"39","author":"Lin","year":"2024","journal-title":"Virol Sin"},{"key":"B2","doi-asserted-by":"publisher","DOI":"10.1128\/mra.00260-24","article-title":"Detection and genomic characterization of an avian influenza virus A\/mute swan\/Mangystau\/1-S24R-2\/2024 (H5N1; clade 2.3.4.4b) strain isolated from the lung of a dead swan in Kazakhstan","volume":"22","author":"Tabynov","year":"2024","journal-title":"Microbiol Resour Announc"},{"key":"B3","doi-asserted-by":"publisher","DOI":"10.3201\/eid2808.212241","article-title":"Novel reassortant avian influenza A(H5N6) virus, China, 2021","volume":"28","author":"Chen","year":"2022","journal-title":"Emerg Infect Dis"},{"key":"B4","doi-asserted-by":"publisher","DOI":"10.1126\/science.abg6302","article-title":"Emerging H5N8 avian influenza viruses","volume":"372","author":"Shi","year":"2021","journal-title":"Science"},{"key":"B5","doi-asserted-by":"publisher","DOI":"10.3201\/eid2710.210297","article-title":"Risk assessment for highly pathogenic avian influenza A(H5N6\/H5N8) clade 2.3.4.4 viruses","volume":"27","author":"Bui","year":"2021","journal-title":"Emerg Infect Dis"},{"key":"B6","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines11091397","article-title":"Immunogenicity and cross-protective efficacy induced by an inactivated recombinant avian influenza A\/H5N1 (Clade 2.3.4.4b) vaccine against co-circulating influenza A\/H5Nx viruses","volume":"11","author":"Mahmoud","year":"2023","journal-title":"Vaccines (Basel)"},{"key":"B7","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2021.1872355","article-title":"Emergence and spread of novel H5N8, H5N5 and H5N1 clade 2.3.4.4 highly pathogenic avian influenza in 2020","volume":"10","author":"Lewis","year":"2021","journal-title":"Emerg Microbes Infect"},{"key":"B8","doi-asserted-by":"publisher","first-page":"e0115122","DOI":"10.1128\/mra.01151-22","article-title":"Genome sequence of highly pathogenic avian influenza virus A\/chicken\/north Kazakhstan\/184\/2020 (H5N8)","volume":"12","author":"Baikara","year":"2023","journal-title":"Microbiol Resour Announc"},{"key":"B9","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.13038","article-title":"Highly pathogenic avian influenza virus of the A\/H5N8 subtype, clade 2.3.4.4b, caused outbreaks in Kazakhstan in 2020","volume":"10","author":"Amirgazin","year":"2022","journal-title":"PeerJ"},{"key":"B10","unstructured":"Before the mass death of swans, sewage water was discharged into the Karakol. What will happen next? Radio Azattyk\n          \n          \n          2024"},{"key":"B11","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines12121448","article-title":"Combination adjuvants enhance recombinant H5 hemagglutinin vaccine protection against high-dose viral challenge in chickens","volume":"12","author":"He","year":"2024","journal-title":"Vaccines (Basel)"},{"key":"B12","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines12060647","article-title":"Evaluation of efficacy of surface coated versus encapsulated influenza antigens in mannose-chitosan nanoparticle-based intranasal vaccine in swine","volume":"12","author":"Bugybayeva","year":"2024","journal-title":"Vaccines (Basel)"},{"key":"B13","unstructured":"Avian influenza prevention: how many birds are vaccinated in Kazakhstan\n          \n          \n          2020"},{"key":"B14","doi-asserted-by":"publisher","DOI":"10.1637\/10183-041012-Review.1","article-title":"Impact of vaccines and vaccination on global control of avian influenza","volume":"56","author":"Swayne","year":"2012","journal-title":"Avian Dis"},{"key":"B15","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00097-12","article-title":"Traditional and new influenza vaccines","volume":"26","author":"Wong","year":"2013","journal-title":"Clin Microbiol Rev"},{"key":"B16","doi-asserted-by":"publisher","DOI":"10.1080\/22221751.2023.2287682","article-title":"A single immunization with H5N1 virus-like particle vaccine protects chickens against divergent H5N1 influenza viruses and vaccine efficacy is determined by adjuvant and dosage","volume":"13","author":"Kong","year":"2024","journal-title":"Emerg Microbes Infect"},{"key":"B17","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines10101683","article-title":"AddaVax-adjuvanted H5N8 inactivated vaccine induces robust humoral immune response against different clades of H5 viruses","volume":"10","author":"Gao","year":"2022","journal-title":"Vaccines (Basel)"},{"key":"B18","doi-asserted-by":"publisher","first-page":"6998","DOI":"10.1016\/j.vaccine.2022.10.018","article-title":"Reverse genetics based H5N2 vaccine provides clinical protection against H5N1, H5N8 and H9N2 avian influenza infection in chickens","volume":"40","author":"Panickan","year":"2022","journal-title":"Vaccine"},{"key":"B19","unstructured":"Summary of status of development and availability of A(H5) non\u2013A(H5N1) candidate vaccine viruses and potency testing reagents\n          \n          \n          \n          2024"},{"key":"B20","doi-asserted-by":"publisher","DOI":"10.3389\/fvets.2022.815978","article-title":"A spike protein-based subunit SARS-coV-2 vaccine for pets: safety, immunogenicity, and protective efficacy in juvenile cats","volume":"9","author":"Tabynov","year":"2022","journal-title":"Front Vet Sci"},{"key":"B21","article-title":"A simple method for estimating fifty percent endpoints","volume":"27","author":"Reed","year":"1938","journal-title":"Am J Hyg"},{"key":"B22","doi-asserted-by":"publisher","first-page":"12485","DOI":"10.1038\/s41598-023-39818-8","article-title":"Author Correction: An intranasal vaccine comprising SARS\u2212CoV\u22122 spike receptor\u2212binding domain protein entrapped in mannose\u2212conjugated chitosan nanoparticle provides protection in hamsters","volume":"13","author":"Tabynov","year":"2023","journal-title":"Sci Rep"},{"key":"B23","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2022.828690","article-title":"Evaluation of a novel adjuvanted vaccine for ultrashort regimen therapy of artemisia pollen-induced allergic bronchial asthma in a mouse model","volume":"13","author":"Tabynov","year":"2022","journal-title":"Front Immunol"},{"key":"B24","doi-asserted-by":"publisher","DOI":"10.3791\/55833","article-title":"An optimized hemagglutination inhibition (HI) assay to quantify influenza-specific antibody titers","author":"Kaufmann","year":"2017","journal-title":"J Vis Exp"},{"key":"B25","article-title":"WHO animal influenza manual","volume":"2002","author":"Webster","year":"2002","journal-title":"WHO\/CDS\/CSR\/NCS"},{"key":"B26","doi-asserted-by":"publisher","first-page":"24","DOI":"10.1038\/s41541-022-00450-8","article-title":"An adjuvanted subunit SARS-CoV-2 spike protein vaccine provides protection against Covid-19 infection and transmission","volume":"7","author":"Tabynov","year":"2022","journal-title":"NPJ Vaccines"},{"key":"B27","doi-asserted-by":"publisher","DOI":"10.2307\/1587638","article-title":"Body temperatures of immature White Leghorn cockerels infected with Newcastle disease virus","volume":"5","author":"Squibb","year":"1961","journal-title":"Avian Dis"},{"key":"B28","doi-asserted-by":"publisher","DOI":"10.1016\/j.rvsc.2022.03.010","article-title":"Tissue tropism and pathology of highly pathogenic avian influenza H5N6 virus in chickens and Pekin ducks","author":"Vreman","year":"2022","journal-title":"Res Vet Sci"},{"key":"B29","doi-asserted-by":"publisher","DOI":"10.1177\/0300985813485099","article-title":"Principles for valid histopathologic scoring in research","volume":"50","author":"Gibson-Corley","year":"2013","journal-title":"Vet Pathol"},{"key":"B30","doi-asserted-by":"publisher","DOI":"10.1016\/j.micpath.2016.06.011","article-title":"Reverse genetics based rgH5N2 vaccine provides protection against high dose challenge of H5N1 avian influenza virus in chicken","volume":"97","author":"Bhatia","year":"2016","journal-title":"Microb Pathog"},{"key":"B31","unstructured":"Healthcare & Pharmaceuticals Public Health. Bird flu vaccination policies by country\n          \n          \n          \n          2023"},{"key":"B32","doi-asserted-by":"publisher","DOI":"10.1016\/j.biologicals.2005.08.003","article-title":"Overview of avian influenza DIVA test strategies","volume":"33","author":"Suarez","year":"2005","journal-title":"Biologicals"},{"key":"B33","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1016\/j.coviro.2020.06.001","article-title":"Reverse genetics approaches for the development of new vaccines against influenza A virus infections","volume":"44","author":"Martinez-Sobrido","year":"2020","journal-title":"Curr Opin Virol"},{"key":"B34","doi-asserted-by":"publisher","first-page":"9697","DOI":"10.1038\/s41598-018-28057-x","article-title":"Efficacy of commercial vaccines against newly emerging avian influenza H5N8 virus in Egypt","volume":"8","author":"Kandeil","year":"2018","journal-title":"Sci Rep"},{"key":"B35","doi-asserted-by":"publisher","DOI":"10.31083\/j.fbl2709268","article-title":"Evaluation of efficacy of oil adjuvanted H5N6 inactivated vaccine against highly pathogenic H5N6 and H5N1 influenza viruses infected chickens","volume":"27","author":"Kuruppuarachchi","year":"2022","journal-title":"Front Biosci (Landmark Ed)"},{"key":"B36","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.micro.51.1.311","article-title":"Interaction of antigens and antibodies at mucosal surfaces","volume":"51","author":"Lamm","year":"1997","journal-title":"Annu Rev Microbiol"},{"key":"B37","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2010.12.013","article-title":"Single dose of oil-adjuvanted inactivated vaccine protects chickens from lethal infections of highly pathogenic H5N1 influenza virus","volume":"29","author":"Hwang","year":"2011","journal-title":"Vaccine"},{"key":"B38","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2023.08.009","article-title":"An Advax-CpG adjuvanted recombinant H5 hemagglutinin vaccine protects mice against lethal influenza infection","volume":"41","author":"Honda-Okubo","year":"2023","journal-title":"Vaccine"}],"container-title":["Frontiers in Immunology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1539492\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,3,20]],"date-time":"2025-03-20T08:00:34Z","timestamp":1742457634000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fimmu.2025.1539492\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,20]]},"references-count":38,"alternative-id":["10.3389\/fimmu.2025.1539492"],"URL":"https:\/\/doi.org\/10.3389\/fimmu.2025.1539492","relation":{},"ISSN":["1664-3224"],"issn-type":[{"value":"1664-3224","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,20]]},"article-number":"1539492"}}