{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T09:44:00Z","timestamp":1774518240878,"version":"3.50.1"},"reference-count":64,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T00:00:00Z","timestamp":1774483200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T00:00:00Z","timestamp":1774483200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"name":"The Finnish Diabetes Research Foundation, Finland"},{"name":"Paulo Foundation, Finland"},{"name":"Instrumentarium Science Foundation, Finland"},{"name":"Tampere Tuberculosis Foundation, Finland"},{"name":"Tampere University graduate school, Finland"},{"name":"The Finnish Foundation for Cardiovascular Research , Finland"},{"name":"Jane and Aatos Erkko Foundation, Finland","award":["240002"],"award-info":[{"award-number":["240002"]}]},{"name":"The Finnish Cultural Foundation, Finland"},{"DOI":"10.13039\/501100014438","name":"Business Finland","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100014438","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002341","name":"Research Council of Finland","doi-asserted-by":"publisher","award":["#369146"],"award-info":[{"award-number":["#369146"]}],"id":[{"id":"10.13039\/501100002341","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002341","name":"Research Council of Finland","doi-asserted-by":"publisher","award":["#309455"],"award-info":[{"award-number":["#309455"]}],"id":[{"id":"10.13039\/501100002341","id-type":"DOI","asserted-by":"publisher"}]},{"name":"The Competitive State Research Financing of the Expert Responsibility Area of Nordlab , Finland","award":["KT0016"],"award-info":[{"award-number":["KT0016"]}]},{"name":"Fimlab Laboratories, Finland","award":["X51409"],"award-info":[{"award-number":["X51409"]}]},{"name":"Sigrid Juselius Foundation, Finland","award":["95-7202-38"],"award-info":[{"award-number":["95-7202-38"]}]},{"DOI":"10.13039\/501100004012","name":"Jane and Aatos Erkko Foundation","doi-asserted-by":"crossref","award":["240002"],"award-info":[{"award-number":["240002"]}],"id":[{"id":"10.13039\/501100004012","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Tampere Institute of Advanced Studies, Finland"},{"name":"The Finnish Cultural Foundation"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["J Biomed Sci"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Enteroviruses, including coxsackievirus B1 (CVB1), cause severe diseases such as myocarditis and meningitis, but vaccines are lacking for most enteroviruses. Conserved and immunodominant epitopes, such as VP4 region and VP1 N-terminus may limit vaccine efficacy by inducing non-neutralizing antibody responses. Virus-like particles (VLPs) mimic native viruses without genetic material and can be engineered to exclude epitopes. To address these challenges, we developed a CVB1-VLP lacking VP4.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>Sequence conservation of CVB VP4 protein and the VP1 N-terminal PALXA region was assessed, and BALB\/c mice were sequentially immunized with different formalin inactivated CVB vaccines. VLP\u0394VP4 was produced using baculovirus-insect cell expression system, was purified, and characterized by SDS-PAGE, transmission electron microscopy, dynamic light scattering, cryogenic electron microscopy, three-dimensional image reconstruction and atomic modelling. VLP\u0394VP4 stability was monitored over five years at 8\u00a0\u00b0C. Comprehensive preclinical experiments were conducted in mice with VLP\u0394VP4, VLP\u0394palxa and inactivated CVB1. Vaccine immunogenicity was evaluated by neutralization assay, ELISA, ELISpot, and in vitro infection assays.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>VP4- and PALXA-regions were conserved among CVB serotypes and sequential mouse vaccinations confirmed the induction of antibodies against these regions, that should be avoided in vaccination. VLP\u0394VP4 exhibited\u2009&gt;\u200995% purity, expected morphology (~\u200930\u00a0nm), exceptional stability at 8\u00a0\u00b0C for five years, and the atomic modelling to 2.7\u00a0\u00c5 resolution showed that the particles were entirely in expanded form. Excluding VP4 from VLP improved production yield 3.5-fold, enhancing scalability of production. Immunological assays demonstrated that VLP\u0394VP4 induced slightly Th2-skewed response, but including adjuvant system 04 (AS04) in the vaccine induced balanced humoral and cellular immune response in mice. Sera from all vaccine groups modulated CVB1 infection, but IFN-\u03b1 induction was lowest in VLP groups, suggesting reduced risk for antibody dependent enhancement of infection. VLP\u0394VP4 elicited significantly higher IFN-\u03b3 responses compared to other vaccines, indicating robust cellular immune response. Antibody responses were comparable across adjuvanted groups, but inclusion of VP4 in the vaccine correlated with weaker systemic T-cell responses.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>VLP\u0394VP4 represents a promising next-generation CVB vaccine candidate with broad applicability against enteroviruses. Removal of VP4 may mitigate the risk for non-beneficial immune imprinting while enabling high purity, long-term stability, and improved manufacturing efficiency.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Graphical Abstract<\/jats:title>\n                  <\/jats:sec>","DOI":"10.1186\/s12929-026-01229-y","type":"journal-article","created":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T08:30:26Z","timestamp":1774513826000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Coxsackie B1 virus-like particle that lacks VP4 protein demonstrates improved vaccine scalability, stability and immunogenicity"],"prefix":"10.1186","volume":"33","author":[{"given":"Saana","family":"Soppela","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Henna-Maarit","family":"Kyr\u00f6l\u00e4inen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alesia","family":"Levanova","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Magloire Pandoua","family":"Nekoua","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mart\u00edn","family":"Gonz\u00e1lez-Rodr\u00edguez","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heini","family":"Lehto","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kiran L. L.","family":"Ahmad","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sergey","family":"Guryanov","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vesa P.","family":"Hyt\u00f6nen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Olli H.","family":"Laitinen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ilkka S.","family":"Junttila","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Didier","family":"Hober","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sarah J.","family":"Butcher","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Minna M.","family":"Hankaniemi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2026,3,26]]},"reference":[{"key":"1229_CR1","unstructured":"International Committee on Taxonomy of Viruses (ICTV). Taxonomy of viruses. (Report, 2024). https:\/\/ictv.global\/"},{"key":"1229_CR2","unstructured":"The Pirbright Institute, U. K. The Picornavirus pages. 2019. https:\/\/www.picornaviridae.com\/. Accessed 11 Nov 2025."},{"key":"1229_CR3","doi-asserted-by":"publisher","first-page":"297","DOI":"10.5005\/jp-journals-11002-0036","volume":"1","author":"S Singh","year":"2022","unstructured":"Singh S, et al. Enteroviral infections in infants. Newborn Clarksville Md. 2022;1:297\u2013305.\u00a0https:\/\/doi.org\/10.5005\/jp-journals-11002-0036","journal-title":"Newborn Clarksville Md"},{"key":"1229_CR4","doi-asserted-by":"publisher","DOI":"10.1186\/s12929-024-01058-x","volume":"31","author":"M Jartti","year":"2024","unstructured":"Jartti M, Flodstr\u00f6m-Tullberg M, Hankaniemi MM. Enteroviruses: epidemic potential, challenges and opportunities with vaccines. J Biomed Sci. 2024;31:73. https:\/\/doi.org\/10.1186\/s12929-024-01058-x.","journal-title":"J Biomed Sci"},{"key":"1229_CR5","doi-asserted-by":"publisher","unstructured":"Bouin A, et al. Major persistent 5\u2032 terminally deleted coxsackievirus B3 populations in human endomyocardial tissues. Emerg Infect Dis J CDC. 2016;22(8). https:\/\/doi.org\/10.3201\/eid2208.160186.","DOI":"10.3201\/eid2208.160186"},{"key":"1229_CR6","doi-asserted-by":"publisher","first-page":"737","DOI":"10.1210\/endrev\/bnad007","volume":"44","author":"A Carr\u00e9","year":"2023","unstructured":"Carr\u00e9 A, Vecchio F, Flodstr\u00f6m-Tullberg M, You S, Mallone R. Coxsackievirus and type 1 diabetes: diabetogenic mechanisms and implications for prevention. Endocr Rev. 2023;44:737\u201351. https:\/\/doi.org\/10.1210\/endrev\/bnad007.","journal-title":"Endocr Rev"},{"issue":"11","key":"1229_CR7","doi-asserted-by":"publisher","first-page":"2491","DOI":"10.1007\/s00125-021-05525-0","volume":"64","author":"S Oikarinen","year":"2021","unstructured":"Oikarinen S, et al. Characterisation of enterovirus RNA detected in the pancreas and other specimens of live patients with newly diagnosed type 1 diabetes in the DiViD study. Diabetologia. 2021;64(11):2491\u2013501. https:\/\/doi.org\/10.1007\/s00125-021-05525-0.","journal-title":"Diabetologia"},{"key":"1229_CR8","doi-asserted-by":"publisher","first-page":"1226","DOI":"10.1007\/s00125-025-06401-x","volume":"68","author":"SJ Richardson","year":"2025","unstructured":"Richardson SJ, et al. Joint analysis of the nPOD-Virus Group data: the association of enterovirus with type 1 diabetes is supported by multiple markers of infection in pancreas tissue. Diabetologia. 2025;68:1226\u201341. https:\/\/doi.org\/10.1007\/s00125-025-06401-x.","journal-title":"Diabetologia"},{"key":"1229_CR9","doi-asserted-by":"publisher","first-page":"919","DOI":"10.3390\/v12090919","volume":"12","author":"M Glenet","year":"2020","unstructured":"Glenet M, et al. Structures and functions of viral 5\u2032 non-coding genomic RNA domain-I in group-B enterovirus infections. Viruses. 2020;12:919.\u00a0https:\/\/doi.org\/10.3390\/v12090919","journal-title":"Viruses"},{"key":"1229_CR10","doi-asserted-by":"publisher","DOI":"10.1002\/rmv.70004","author":"CJ Mbani","year":"2024","unstructured":"Mbani CJ, et al. Enterovirus antibodies: friends and foes. Rev Med Virol. 2024. https:\/\/doi.org\/10.1002\/rmv.70004.","journal-title":"Rev Med Virol"},{"key":"1229_CR11","doi-asserted-by":"publisher","first-page":"368","DOI":"10.1038\/s41579-018-0005-4","volume":"16","author":"J Baggen","year":"2018","unstructured":"Baggen J, Thibaut HJ, Strating JRPM, van Kuppeveld FJM. The life cycle of non-polio enteroviruses and how to target it. Nat Rev Microbiol. 2018;16:368\u201381. https:\/\/doi.org\/10.1038\/s41579-018-0005-4.","journal-title":"Nat Rev Microbiol"},{"key":"1229_CR12","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.00082-22","volume":"96","author":"JT Kelly","year":"2022","unstructured":"Kelly JT, et al. Membrane interactions and uncoating of Aichi virus, a picornavirus that lacks a VP4. J Virol. 2022;96:e00082-22. https:\/\/doi.org\/10.1128\/jvi.00082-22.","journal-title":"J Virol"},{"key":"1229_CR13","doi-asserted-by":"publisher","first-page":"448","DOI":"10.1016\/j.chom.2021.01.001","volume":"29","author":"L Xu","year":"2021","unstructured":"Xu L, et al. Cryo-EM structures reveal the molecular basis of receptor-initiated coxsackievirus uncoating. Cell Host Microbe. 2021;29:448-462.e5. https:\/\/doi.org\/10.1016\/j.chom.2021.01.001.","journal-title":"Cell Host Microbe"},{"key":"1229_CR14","doi-asserted-by":"publisher","DOI":"10.3389\/fcimb.2023.1216364","author":"V Lampinen","year":"2023","unstructured":"Lampinen V, et al. SpyTag\/SpyCatcher display of influenza M2e peptide on norovirus-like particle provides stronger immunization than direct genetic fusion. Front Cell Infect Microbiol. 2023. https:\/\/doi.org\/10.3389\/fcimb.2023.1216364.","journal-title":"Front Cell Infect Microbiol"},{"key":"1229_CR15","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-024-65316-6","volume":"14","author":"V Lampinen","year":"2024","unstructured":"Lampinen V, et al. Production of norovirus-, rotavirus-, and enterovirus-like particles in insect cells is simplified by plasmid-based expression. Sci Rep. 2024;14:14874. https:\/\/doi.org\/10.1038\/s41598-024-65316-6.","journal-title":"Sci Rep"},{"key":"1229_CR16","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/j.smim.2017.08.014","volume":"34","author":"MO Mohsen","year":"2017","unstructured":"Mohsen MO, Zha L, Cabral-Miranda G, Bachmann MF. Major findings and recent advances in virus-like particle (VLP)-based vaccines. Semin Immunol. 2017;34:123\u201332. https:\/\/doi.org\/10.1016\/j.smim.2017.08.014.","journal-title":"Semin Immunol"},{"key":"1229_CR17","doi-asserted-by":"publisher","DOI":"10.1186\/s12929-025-01183-1","volume":"32","author":"S Soppela","year":"2025","unstructured":"Soppela S, et al. Coxsackie B1 virus-like particle vaccine modified to exclude a highly conserved immunoreactive region from the capsid induces potent neutralizing antibodies and protects against infection in mice. J Biomed Sci. 2025;32:86. https:\/\/doi.org\/10.1186\/s12929-025-01183-1.","journal-title":"J Biomed Sci"},{"key":"1229_CR18","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-025-94656-0","volume":"15","author":"S Soppela","year":"2025","unstructured":"Soppela S, et al. Immunological and structural evaluation of the intranasally administrated CVB1 whole-virus and VLP vaccines. Sci Rep. 2025;15:10198. https:\/\/doi.org\/10.1038\/s41598-025-94656-0.","journal-title":"Sci Rep"},{"key":"1229_CR19","first-page":"572","volume":"104","author":"T Francis","year":"1960","unstructured":"Francis T. On the doctrine of original antigenic sin. Proc Am Philos Soc. 1960;104:572\u20138.\u00a0https:\/\/www.jstor.org\/stable\/985534","journal-title":"Proc Am Philos Soc"},{"key":"1229_CR20","doi-asserted-by":"publisher","DOI":"10.1080\/21645515.2024.2384192","volume":"20","author":"M Maltseva","year":"2024","unstructured":"Maltseva M, Keeshan A, Cooper C, Langlois M-A. Immune imprinting: the persisting influence of the first antigenic encounter with rapidly evolving viruses. Hum Vaccin Immunother. 2024;20:2384192. https:\/\/doi.org\/10.1080\/21645515.2024.2384192.","journal-title":"Hum Vaccin Immunother"},{"key":"1229_CR21","doi-asserted-by":"publisher","unstructured":"Mishra N, et al. Antibodies to enteroviruses in cerebrospinal fluid of patients with acute flaccid myelitis. mBio. 2019;10. https:\/\/doi.org\/10.1128\/mbio.01903-19.","DOI":"10.1128\/mbio.01903-19"},{"key":"1229_CR22","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1111\/j.1365-2249.2011.04559.x","volume":"168","author":"D Hober","year":"2012","unstructured":"Hober D, et al. Immunology in the clinic review series; focus on type 1 diabetes and viruses: role of antibodies enhancing the infection with Coxsackievirus-B in the pathogenesis of type 1 diabetes. Clin Exp Immunol. 2012;168:47\u201351. https:\/\/doi.org\/10.1111\/j.1365-2249.2011.04559.x.","journal-title":"Clin Exp Immunol"},{"key":"1229_CR23","doi-asserted-by":"publisher","DOI":"10.1128\/mmbr.00240","volume":"0","author":"C Morvan","year":"2025","unstructured":"Morvan C, Nekoua MP, Debuysschere C, Alidjinou EK, Hober D. Enhancement of viral infection by antibodies and consequences. Microbiol Mol Biol Rev. 2025;0:e00240-25. https:\/\/doi.org\/10.1128\/mmbr.00240.","journal-title":"Microbiol Mol Biol Rev"},{"key":"1229_CR24","doi-asserted-by":"publisher","unstructured":"Samuelson A, Forsgren M, Johansson BO, Wahren B, Sallberg M. Molecular basis for serological cross-reactivity between enteroviruses. Clin Diagn Lab Immunol. 1994;1(3):336\u201341. https:\/\/doi.org\/10.1128\/cdli.1.3.336-341.1994","DOI":"10.1128\/cdli.1.3.336-341.1994"},{"key":"1229_CR25","doi-asserted-by":"publisher","first-page":"1919","DOI":"10.1099\/0022-1317-80-8-1919","volume":"80","author":"A Airaksinen","year":"1999","unstructured":"Airaksinen A, Roivainen M, Stanway G, Hovi T. Site-saturation mutagenesis of the PALTAVETG motif in coxsackievirus A9 capsid protein VP1 reveals evidence of conservation of a periodic hydrophobicity profile. J Gen Virol. 1999;80:1919\u201327. https:\/\/doi.org\/10.1099\/0022-1317-80-8-1919.","journal-title":"J Gen Virol"},{"key":"1229_CR26","doi-asserted-by":"publisher","first-page":"3028","DOI":"10.1128\/jcm.41.7.3028-3034.2003","volume":"41","author":"S-Y Shin","year":"2003","unstructured":"Shin S-Y, et al. Identification of enteroviruses by using monoclonal antibodies against a putative common epitope. J Clin Microbiol. 2003;41:3028\u201334. https:\/\/doi.org\/10.1128\/jcm.41.7.3028-3034.2003.","journal-title":"J Clin Microbiol"},{"key":"1229_CR27","doi-asserted-by":"publisher","first-page":"e1011223","DOI":"10.1371\/journal.ppat.1011223","volume":"19","author":"A Teo","year":"2023","unstructured":"Teo A, Tan HD, Loy T, Chia PY, Chua CLL. Understanding antibody-dependent enhancement in dengue: are afucosylated IgG1s a concern? PLoS Pathog. 2023;19:e1011223.\u00a0https:\/\/doi.org\/10.1371\/journal.ppat.1011223","journal-title":"PLoS Pathog"},{"key":"1229_CR28","doi-asserted-by":"publisher","first-page":"175","DOI":"10.1126\/science.aal4365","volume":"356","author":"SV Bardina","year":"2017","unstructured":"Bardina SV, et al. Enhancement of Zika virus pathogenesis by preexisting antiflavivirus immunity. Science. 2017;356:175\u201380. https:\/\/doi.org\/10.1126\/science.aal4365.","journal-title":"Science"},{"key":"1229_CR29","doi-asserted-by":"publisher","DOI":"10.1017\/cts.2020.39","author":"J Wang","year":"2020","unstructured":"Wang J, Zand MS. The potential for antibody-dependent enhancement of SARS-CoV-2 infection: translational implications for vaccine development. J Clin Transl Sci. 2020. https:\/\/doi.org\/10.1017\/cts.2020.39.","journal-title":"J Clin Transl Sci"},{"key":"1229_CR30","doi-asserted-by":"publisher","first-page":"3910","DOI":"10.1016\/j.vaccine.2017.06.004","volume":"35","author":"RA Mart\u00ednez-Vega","year":"2017","unstructured":"Mart\u00ednez-Vega RA, Carrasquila G, Luna E, Ramos-Casta\u00f1eda J. ADE and dengue vaccination. Vaccine. 2017;35:3910\u20132. https:\/\/doi.org\/10.1016\/j.vaccine.2017.06.004.","journal-title":"Vaccine"},{"key":"1229_CR31","doi-asserted-by":"publisher","first-page":"13882","DOI":"10.1128\/JVI.79.22.13882-13891.2005","volume":"79","author":"W Chehadeh","year":"2005","unstructured":"Chehadeh W, et al. Viral protein VP4 is a target of human antibodies enhancing Coxsackievirus B4- and B3-induced synthesis of alpha interferon. J Virol. 2005;79:13882\u201391. https:\/\/doi.org\/10.1128\/JVI.79.22.13882-13891.2005.","journal-title":"J Virol"},{"key":"1229_CR32","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.29399","volume":"96","author":"C Morvan","year":"2024","unstructured":"Morvan C, Nekoua MP, Debuysschere C, Alidjinou EK, Hober D. Antibody-dependent enhancement and neutralization against CVB4 investigated in vitro and in silico through an agent-based model. J Med Virol. 2024;96:e29399. https:\/\/doi.org\/10.1002\/jmv.29399.","journal-title":"J Med Virol"},{"key":"1229_CR33","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1016\/j.virusres.2007.01.001","volume":"125","author":"P Sauter","year":"2007","unstructured":"Sauter P, et al. Role of the capsid protein VP4 in the plasma-dependent enhancement of the Coxsackievirus B4E2-infection of human peripheral blood cells. Virus Res. 2007;125:183\u201390. https:\/\/doi.org\/10.1016\/j.virusres.2007.01.001.","journal-title":"Virus Res"},{"key":"1229_CR34","first-page":"153","volume":"12","author":"A Goffard","year":"2011","unstructured":"Goffard A, Sane F, Soumillon M, Hober D. Specificity of the Coxsackievirus B4 VP4 capsid protein investigated in silico. Discov Med. 2011;12:153\u20138.\u00a0https:\/\/europepmc.org\/article\/MED\/21878192","journal-title":"Discov Med"},{"key":"1229_CR35","doi-asserted-by":"publisher","first-page":"866","DOI":"10.1002\/jmv.21171","volume":"80","author":"P Sauter","year":"2008","unstructured":"Sauter P, et al. A part of the VP4 capsid protein exhibited by coxsackievirus B4 E2 is the target of antibodies contained in plasma from patients with type 1 diabetes. J Med Virol. 2008;80:866\u201378. https:\/\/doi.org\/10.1002\/jmv.21171.","journal-title":"J Med Virol"},{"key":"1229_CR36","doi-asserted-by":"publisher","first-page":"11955","DOI":"10.1128\/JVI.01809-14","volume":"88","author":"T Wilton","year":"2014","unstructured":"Wilton T, Dunn G, Eastwood D, Minor PD, Martin J. Effect of formaldehyde inactivation on poliovirus. J Virol. 2014;88:11955\u201364. https:\/\/doi.org\/10.1128\/JVI.01809-14.","journal-title":"J Virol"},{"key":"1229_CR37","doi-asserted-by":"publisher","first-page":"1279","DOI":"10.1016\/j.chom.2022.08.001","volume":"30","author":"Q Zheng","year":"2022","unstructured":"Zheng Q, et al. Structural basis for the synergistic neutralization of coxsackievirus B1 by a triple-antibody cocktail. Cell Host Microbe. 2022;30:1279-1294.e6. https:\/\/doi.org\/10.1016\/j.chom.2022.08.001.","journal-title":"Cell Host Microbe"},{"key":"1229_CR38","doi-asserted-by":"publisher","first-page":"1412","DOI":"10.1002\/jmv.23903","volume":"86","author":"S H\u00e4m\u00e4l\u00e4inen","year":"2014","unstructured":"H\u00e4m\u00e4l\u00e4inen S, et al. Coxsackievirus B1 reveals strain specific differences in plasmacytoid dendritic cell mediated immunogenicity. J Med Virol. 2014;86:1412\u201320. https:\/\/doi.org\/10.1002\/jmv.23903.","journal-title":"J Med Virol"},{"key":"1229_CR39","doi-asserted-by":"publisher","first-page":"5962","DOI":"10.1016\/j.vaccine.2019.08.037","volume":"37","author":"MM Hankaniemi","year":"2019","unstructured":"Hankaniemi MM, et al. A comparative study of the effect of UV and formalin inactivation on the stability and immunogenicity of a Coxsackievirus B1 vaccine. Vaccine. 2019;37:5962\u201371. https:\/\/doi.org\/10.1016\/j.vaccine.2019.08.037.","journal-title":"Vaccine"},{"key":"1229_CR40","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.aaz2433","author":"VM Stone","year":"2020","unstructured":"Stone VM, et al. A hexavalent Coxsackievirus B vaccine is highly immunogenic and has a strong protective capacity in mice and nonhuman primates. Sci Adv. 2020. https:\/\/doi.org\/10.1126\/sciadv.aaz2433.","journal-title":"Sci Adv"},{"key":"1229_CR41","doi-asserted-by":"publisher","DOI":"10.1016\/j.antiviral.2019.104595","volume":"171","author":"MM Hankaniemi","year":"2019","unstructured":"Hankaniemi MM, et al. Formalin treatment increases the stability and immunogenicity of coxsackievirus B1 VLP vaccine. Antivir Res. 2019;171:104595. https:\/\/doi.org\/10.1016\/j.antiviral.2019.104595.","journal-title":"Antivir Res"},{"key":"1229_CR42","doi-asserted-by":"publisher","first-page":"290","DOI":"10.1038\/nmeth.4169","volume":"14","author":"A Punjani","year":"2017","unstructured":"Punjani A, Rubinstein JL, Fleet DJ, Brubaker MA. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat Methods. 2017;14:290\u20136. https:\/\/doi.org\/10.1038\/nmeth.4169.","journal-title":"Nat Methods"},{"key":"1229_CR43","doi-asserted-by":"publisher","first-page":"721","DOI":"10.1016\/j.jmb.2003.07.013","volume":"333","author":"PB Rosenthal","year":"2003","unstructured":"Rosenthal PB, Henderson R. Optimal determination of particle orientation, absolute hand, and contrast loss in single-particle electron cryomicroscopy. J Mol Biol. 2003;333:721\u201345. https:\/\/doi.org\/10.1016\/j.jmb.2003.07.013.","journal-title":"J Mol Biol"},{"key":"1229_CR44","doi-asserted-by":"publisher","DOI":"10.1002\/pro.4792","volume":"32","author":"EC Meng","year":"2023","unstructured":"Meng EC, et al. UCSF ChimeraX: tools for structure building and analysis. Protein Sci. 2023;32:e4792. https:\/\/doi.org\/10.1002\/pro.4792.","journal-title":"Protein Sci"},{"key":"1229_CR45","doi-asserted-by":"publisher","first-page":"14","DOI":"10.1002\/pro.3235","volume":"27","author":"TD Goddard","year":"2018","unstructured":"Goddard TD, et al. UCSF ChimeraX: meeting modern challenges in visualization and analysis. Protein Sci. 2018;27:14\u201325. https:\/\/doi.org\/10.1002\/pro.3235.","journal-title":"Protein Sci"},{"key":"1229_CR46","doi-asserted-by":"publisher","first-page":"70","DOI":"10.1002\/pro.3943","volume":"30","author":"EF Pettersen","year":"2021","unstructured":"Pettersen EF, et al. UCSF ChimeraX: structure visualization for researchers, educators, and developers. Protein Sci. 2021;30:70\u201382. https:\/\/doi.org\/10.1002\/pro.3943.","journal-title":"Protein Sci"},{"key":"1229_CR47","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1107\/S2059798318002425","volume":"74","author":"TI Croll","year":"2018","unstructured":"Croll TI. ISOLDE: a physically realistic environment for model building into low-resolution electron-density maps. Acta Crystallogr Sect D Struct Biol. 2018;74:519\u201330. https:\/\/doi.org\/10.1107\/S2059798318002425.","journal-title":"Acta Crystallogr Sect D Struct Biol"},{"key":"1229_CR48","doi-asserted-by":"publisher","first-page":"861","DOI":"10.1107\/S2059798319011471","volume":"75","author":"D Liebschner","year":"2019","unstructured":"Liebschner D, et al. Macromolecular structure determination using X-rays, neutrons and electrons: recent developments in Phenix. Acta Crystallogr Sect Struct Biol. 2019;75:861\u201377. https:\/\/doi.org\/10.1107\/S2059798319011471.","journal-title":"Acta Crystallogr Sect Struct Biol"},{"key":"1229_CR49","doi-asserted-by":"publisher","DOI":"10.3390\/microorganisms8091287","volume":"8","author":"MM Hankaniemi","year":"2020","unstructured":"Hankaniemi MM, et al. Structural insight into CVB3-VLP non-adjuvanted vaccine. Microorganisms. 2020;8:10.3390\/microorganisms8091287. https:\/\/doi.org\/10.3390\/microorganisms8091287.","journal-title":"Microorganisms"},{"key":"1229_CR50","doi-asserted-by":"publisher","first-page":"W521","DOI":"10.1093\/nar\/gkae241","volume":"52","author":"F Madeira","year":"2024","unstructured":"Madeira F, et al. The EMBL-EBI job dispatcher sequence analysis tools framework in 2024. Nucleic Acids Res. 2024;52:W521\u20135. https:\/\/doi.org\/10.1093\/nar\/gkae241.","journal-title":"Nucleic Acids Res"},{"key":"1229_CR51","doi-asserted-by":"publisher","first-page":"34","DOI":"10.1002\/jps.22267","volume":"100","author":"LA Brito","year":"2011","unstructured":"Brito LA, Singh M. Acceptable levels of endotoxin in vaccine formulations during preclinical research. J Pharm Sci. 2011;100:34\u20137. https:\/\/doi.org\/10.1002\/jps.22267.","journal-title":"J Pharm Sci"},{"key":"1229_CR52","unstructured":"CBER. Guidance for industry, characterization and qualification of cell substrates and other biological materials used in the production of viral vaccines for infectious disease indications. 2010. https:\/\/www.federalregister.gov\/documents\/2010\/03\/04\/2010-4553\/guidance-for-industry-characterization-and-qualification-of-cell-substrates-and-other-biological"},{"key":"1229_CR53","doi-asserted-by":"publisher","first-page":"17144","DOI":"10.1021\/acs.jmedchem.4c00701","volume":"67","author":"C Tammaro","year":"2024","unstructured":"Tammaro C, et al. SAR analysis of novel Coxsackie virus A9 capsid binders. J Med Chem. 2024;67:17144\u201361. https:\/\/doi.org\/10.1021\/acs.jmedchem.4c00701.","journal-title":"J Med Chem"},{"key":"1229_CR54","doi-asserted-by":"publisher","DOI":"10.1128\/jvi.01367-22","volume":"96","author":"A Domanska","year":"2022","unstructured":"Domanska A, et al. Structural studies reveal that endosomal cations promote formation of infectious Coxsackievirus A9 A-particles, facilitating RNA and VP4 release. J Virol. 2022;96:e0136722. https:\/\/doi.org\/10.1128\/jvi.01367-22.","journal-title":"J Virol"},{"key":"1229_CR55","doi-asserted-by":"publisher","first-page":"7509","DOI":"10.1016\/j.vaccine.2019.09.072","volume":"37","author":"S Heinim\u00e4ki","year":"2019","unstructured":"Heinim\u00e4ki S, et al. Combination of three virus-derived nanoparticles as a vaccine against enteric pathogens; enterovirus, norovirus and rotavirus. Vaccine. 2019;37:7509\u201318. https:\/\/doi.org\/10.1016\/j.vaccine.2019.09.072.","journal-title":"Vaccine"},{"key":"1229_CR56","doi-asserted-by":"publisher","first-page":"527","DOI":"10.1080\/21505594.2016.1152442","volume":"7","author":"F Elmastour","year":"2016","unstructured":"Elmastour F, et al. Immunoglobulin G-dependent enhancement of the infection with Coxsackievirus B4 in a murine system. Virulence. 2016;7:527\u201335. https:\/\/doi.org\/10.1080\/21505594.2016.1152442.","journal-title":"Virulence"},{"key":"1229_CR57","doi-asserted-by":"publisher","first-page":"156","DOI":"10.1177\/014107680609900320","volume":"99","author":"M Fitzpatrick","year":"2006","unstructured":"Fitzpatrick M. The cutter incident: how America\u2019s first polio vaccine led to a growing vaccine crisis. J R Soc Med. 2006;99:156.\u00a0https:\/\/pmc.ncbi.nlm.nih.gov\/articles\/PMC1410842\/","journal-title":"J R Soc Med"},{"key":"1229_CR58","doi-asserted-by":"publisher","unstructured":"Wang X, et al. Structure, immunogenicity, and protective mechanism of an engineered enterovirus 71-like particle vaccine mimicking 80s empty capsid. J Virol. 2017;92. https:\/\/doi.org\/10.1128\/jvi.01330-17.","DOI":"10.1128\/jvi.01330-17"},{"key":"1229_CR59","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1007\/978-1-4939-3292-4_15","volume":"1387","author":"T Wilton","year":"2016","unstructured":"Wilton T. Methods for the quality control of inactivated poliovirus vaccines. Methods Mol Biol. 2016;1387:279\u201397. https:\/\/doi.org\/10.1007\/978-1-4939-3292-4_15.","journal-title":"Methods Mol Biol"},{"key":"1229_CR60","doi-asserted-by":"publisher","DOI":"10.3390\/v16040573","volume":"16","author":"A Yang","year":"2024","unstructured":"Yang A, et al. Quantitation of enterovirus A71 empty and full particles by sedimentation velocity analytical ultracentrifugation. Viruses. 2024;16:573. https:\/\/doi.org\/10.3390\/v16040573.","journal-title":"Viruses"},{"key":"1229_CR61","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines13040404","volume":"13","author":"X Zhang","year":"2025","unstructured":"Zhang X, et al. Development of in vitro potency methods to replace in vivo tests for enterovirus 71 inactivated vaccine (human diploid cell-based\/vero cell-based). Vaccines. 2025;13:404. https:\/\/doi.org\/10.3390\/vaccines13040404.","journal-title":"Vaccines"},{"key":"1229_CR62","doi-asserted-by":"publisher","DOI":"10.1177\/2515135519888998","volume":"7","author":"HX Lim","year":"2019","unstructured":"Lim HX, Poh CL. Insights into innate and adaptive immune responses in vaccine development against EV-A71. Ther Adv Vaccines Immunother. 2019;7:2515135519888998. https:\/\/doi.org\/10.1177\/2515135519888998.","journal-title":"Ther Adv Vaccines Immunother"},{"key":"1229_CR63","doi-asserted-by":"publisher","first-page":"466","DOI":"10.1203\/01.pdr.0000238247.86041.19","volume":"60","author":"L-Y Chang","year":"2006","unstructured":"Chang L-Y, et al. Status of cellular rather than humoral immunity is correlated with clinical outcome of enterovirus 71. Pediatr Res. 2006;60:466\u201371. https:\/\/doi.org\/10.1203\/01.pdr.0000238247.86041.19.","journal-title":"Pediatr Res"},{"key":"1229_CR64","doi-asserted-by":"publisher","first-page":"5988","DOI":"10.1128\/JVI.79.10.5988-5995.2005","volume":"79","author":"R Wahid","year":"2005","unstructured":"Wahid R, Cannon MJ, Chow M. Virus-specific CD4 + and CD8 + cytotoxic T-cell responses and long-term T-cell memory in individuals vaccinated against polio. J Virol. 2005;79:5988\u201395. https:\/\/doi.org\/10.1128\/JVI.79.10.5988-5995.2005.","journal-title":"J Virol"}],"container-title":["Journal of Biomedical Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12929-026-01229-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12929-026-01229-y","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12929-026-01229-y.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T08:30:34Z","timestamp":1774513834000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1186\/s12929-026-01229-y"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,3,26]]},"references-count":64,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2026,12]]}},"alternative-id":["1229"],"URL":"https:\/\/doi.org\/10.1186\/s12929-026-01229-y","relation":{},"ISSN":["1423-0127"],"issn-type":[{"value":"1423-0127","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,3,26]]},"assertion":[{"value":"9 December 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"17 February 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"26 March 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"All animal experiments in this study were carried out following the Finnish Act on the Protection of Animals Used for Scientific or Educational Purposes (497\/2013) or in compliance with Swedish national laws and the National Institutes of Health guidelines for laboratory animal care. Procedures were approved by the Regional State Administrative Agency, Pirkanmaa, Finland (decision number ESAVI\/1408\/2021) or by the Stockholm Southern Animal Ethics Board. All animal reporting was conducted in accordance with ARRIVE guidelines. All efforts were made to minimize animal suffering and to reduce the number of animals used. The welfare of the animals was monitored throughout the experiments.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"SS, VPH and MMH are inventors of the patent application FI 20253217 belonging to Innofend Oy, titled Engineered Enterovirus-Like particles and uses thereof. The rights to the invention have been transferred to the company, and thus, no competing interests exist. SS and MMH are shareholders of Innofend Oy. The other authors declare that they have no conflicts of interest.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"34"}}