{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T05:08:57Z","timestamp":1779340137856,"version":"3.51.4"},"reference-count":29,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T00:00:00Z","timestamp":1779148800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"State Key Laboratory of Drug Regulatory Science","award":["2025SKLDRS0319"],"award-info":[{"award-number":["2025SKLDRS0319"]}]},{"name":"Beijing Natural Science Foundation","award":["L242148"],"award-info":[{"award-number":["L242148"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The high case fatality rate, cross-species transmission, and ongoing evolution of H5 avian influenza viruses pose an imminent threat of an influenza pandemic, particularly with the currently predominant clade 2.3.4.4b lineage. Existing seasonal influenza vaccines and licensed H5 vaccines provide limited cross-protection against H5 viruses, underscoring an urgent need for the development of broadly protective H5 vaccines. In this study, we analyzed all human-infected H5 hemagglutinin (HA) sequences using bioinformatics approaches and subsequently designed a novel H5 influenza vaccine through algorithm optimization. The predicted structure of this vaccine closely resembles that of the wild-type H5 HA trimer. In animal studies, the algorithm-optimized H5 mRNA vaccine not only induced high levels of neutralizing antibodies against multiple clade 2.3.4.4b H5 viruses but also elicited cross-neutralizing antibodies against clade 2.3.4.4 and clade 2.2.1 H5 viruses, as well as robust cellular immune responses. These findings highlight the potential of algorithm-based approaches in developing broadly protective vaccines against pandemic viruses and suggest that this vaccine candidate could serve as a strategic stockpile for preventing H5 influenza pandemics.<\/jats:p>","DOI":"10.3390\/ijms27104547","type":"journal-article","created":{"date-parts":[[2026,5,19]],"date-time":"2026-05-19T11:53:10Z","timestamp":1779191590000},"page":"4547","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Algorithm-Optimized H5 Influenza mRNA Vaccine Induces Broad Immune Responses"],"prefix":"10.3390","volume":"27","author":[{"given":"Liangliang","family":"Wang","sequence":"first","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhengda","family":"Peng","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-1429-0848","authenticated-orcid":false,"given":"Chenchen","family":"He","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jie","family":"Zhang","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pengju","family":"Yu","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4246-8889","authenticated-orcid":false,"given":"Weijin","family":"Huang","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Youchun","family":"Wang","sequence":"additional","affiliation":[{"name":"Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China"},{"name":"Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming 650118, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1807-314X","authenticated-orcid":false,"given":"Chenyan","family":"Zhao","sequence":"additional","affiliation":[{"name":"Division of HIV\/AIDS and Sex-Transmitted Virus Vaccines, Institute for Biological Product Control, National Institutes for Food and Drug Control (NIFDC), Beijing 102629, China"},{"name":"State Key Laboratory of Drug Regulatory Science, Beijing 102629, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,5,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"792","DOI":"10.1637\/0005-2086-47.s3.792","article-title":"Report on avian influenza in the Eastern Hemisphere during 1997\u20132002","volume":"47","author":"Alexander","year":"2003","journal-title":"Avian Dis."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"102638","DOI":"10.1016\/j.tmaid.2023.102638","article-title":"A comprehensive review of highly pathogenic avian influenza (HPAI) H5N1: An imminent threat at doorstep","volume":"55","author":"Charostad","year":"2023","journal-title":"Travel Med. Infect. Dis."},{"key":"ref_3","unstructured":"World Health Organization (2026, March 10). Cumulative Number of Confirmed Human Cases of Avian Influenza A(H5N1) Reported to WHO, 2003\u20132026. Available online: https:\/\/cdn.who.int\/media\/docs\/default-source\/2021-dha-docs\/cumulative-number-of-confirmed-human-cases-for-avian-influenza-a(h5n1)-reported-to-who--2003-2025.pdf."},{"key":"ref_4","unstructured":"U.S. Centers for Disease Control and Prevention (2026, March 10). A(H5) Bird Flu: Current Situation, Available online: https:\/\/www.cdc.gov\/bird-flu\/situation-summary\/index.html."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1534","DOI":"10.1126\/science.1213362","article-title":"Airborne transmission of influenza A\/H5N1 virus between ferrets","volume":"336","author":"Herfst","year":"2012","journal-title":"Science"},{"key":"ref_6","unstructured":"U.S. Food & Drug (2026, March 19). Vaccines Licensed for Use in the United States, Available online: https:\/\/www.fda.gov\/vaccines-blood-biologics\/vaccines\/vaccines-licensed-use-united-states."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1455","DOI":"10.1007\/s00705-022-05442-6","article-title":"A new clade 2.3.4.4b H5N1 highly pathogenic avian influenza genotype detected in Europe in 2021","volume":"167","author":"Nagy","year":"2022","journal-title":"Arch. Virol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Liang, Y., Nissen, J.N., Krog, J.S., Breum, S., Trebbien, R., Larsen, L.E., and Hjulsager, C.K. (2021). Novel Clade 2.3.4.4b Highly Pathogenic Avian Influenza A H5N8 and H5N5 Viruses in Denmark, 2020. Viruses, 13.","DOI":"10.3390\/v13050886"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"e70557","DOI":"10.1002\/mco2.70557","article-title":"Intranasal Mosaic H1N1 Live Attenuated Influenza Vaccine Elicits Broad Cross-Reactive Immunity and Protection Against Group 1 and 2 Influenza A Viruses","volume":"6","author":"Ma","year":"2025","journal-title":"MedComm"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2131","DOI":"10.1016\/j.chom.2024.10.016","article-title":"A trivalent mucosal vaccine encoding phylogenetically inferred ancestral RBD sequences confers pan-Sarbecovirus protection in mice","volume":"32","author":"Case","year":"2024","journal-title":"Cell Host Microbe"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1411","DOI":"10.1016\/j.immuni.2025.04.015","article-title":"Computationally designed proteins mimic antibody immune evasion in viral evolution","volume":"58","author":"Youssef","year":"2025","journal-title":"Immunity"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1038\/s41573-021-00283-5","article-title":"mRNA vaccines for infectious diseases: Principles, delivery and clinical translation","volume":"20","author":"Chaudhary","year":"2021","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"838","DOI":"10.1038\/s41573-024-01042-y","article-title":"mRNA vaccines for infectious diseases\u2014Advances, challenges and opportunities","volume":"23","author":"Pardi","year":"2024","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"33987","DOI":"10.1038\/srep33987","article-title":"Epigraph: A Vaccine Design Tool Applied to an HIV Therapeutic Vaccine and a Pan-Filovirus Vaccine","volume":"6","author":"Theiler","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_15","first-page":"408","article-title":"Molecular characteristics of the first human case infected with the avian influenza virus H5N1 clade 2.3.4.4b in eastern China","volume":"35","author":"Deng","year":"2024","journal-title":"Jiangsu J. Prev. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate structure prediction of biomolecular interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1002\/pro.3943","article-title":"UCSF ChimeraX: Structure visualization for researchers, educators, and developers","volume":"30","author":"Pettersen","year":"2021","journal-title":"Protein Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1038\/s41467-025-67753-x","article-title":"Emergence of mammalian-adaptive PB2 mutations enhances polymerase activity and pathogenicity of cattle-derived H5N1 influenza A virus","volume":"17","author":"Zhang","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1128","DOI":"10.1126\/science.adt0180","article-title":"A single mutation in bovine influenza H5N1 hemagglutinin switches specificity to human receptors","volume":"386","author":"Lin","year":"2024","journal-title":"Science"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"eado9140","DOI":"10.1126\/sciadv.ado9140","article-title":"Association of poultry vaccination with interspecies transmission and molecular evolution of H5 subtype avian influenza virus","volume":"11","author":"Li","year":"2025","journal-title":"Sci. Adv."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"699","DOI":"10.1016\/j.cell.2025.12.033","article-title":"Can H5N1 avian influenza in dairy cattle be contained in the US?","volume":"189","author":"Pekar","year":"2026","journal-title":"Cell"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"4046","DOI":"10.1038\/s41467-025-59182-7","article-title":"Epitope-optimized vaccine elicits enduring immunity against swine influenza A virus","volume":"16","author":"Madapong","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"20824","DOI":"10.1021\/acsnano.5c03199","article-title":"Epitope-Optimized Influenza Hemagglutinin Nanoparticle Vaccine Provides Broad Cross-Reactive Immunity against H9N2 Influenza Virus","volume":"19","author":"Hao","year":"2025","journal-title":"ACS Nano"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1038\/s41564-024-01903-7","article-title":"Vaccine-induced T cell responses control Orthoflavivirus challenge infection without neutralizing antibodies in humans","volume":"10","author":"Kalimuddin","year":"2025","journal-title":"Nat. Microbiol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2456","DOI":"10.1016\/j.scib.2024.02.041","article-title":"Development of T cell antigen-based human coronavirus vaccines against nAb-escaping SARS-CoV-2 variants","volume":"69","author":"Zhou","year":"2024","journal-title":"Sci. Bull."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.virs.2026.02.005","article-title":"The role of T cells in influenza infection and vaccination","volume":"41","author":"Li","year":"2026","journal-title":"Virol. Sin."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"160","DOI":"10.1038\/s41392-024-01876-3","article-title":"Nonconserved epitopes dominate reverse preexisting T cell immunity in COVID-19 convalescents","volume":"9","author":"Wang","year":"2024","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"10954","DOI":"10.1038\/s41467-025-66431-2","article-title":"Pre-existing cross-reactive immunity to highly pathogenic avian influenza 2.3.4.4b A(H5N1) virus in the United States","volume":"16","author":"Li","year":"2025","journal-title":"Nat. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"308","DOI":"10.1002\/mlf2.12085","article-title":"Protective efficacy of a universal influenza mRNA vaccine against the challenge of H1 and H5 influenza A viruses in mice","volume":"2","author":"Li","year":"2023","journal-title":"mLife"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/10\/4547\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,21]],"date-time":"2026-05-21T04:27:47Z","timestamp":1779337667000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/10\/4547"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,19]]},"references-count":29,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["ijms27104547"],"URL":"https:\/\/doi.org\/10.3390\/ijms27104547","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,19]]}}}