{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,2]],"date-time":"2026-10-02T21:05:33Z","timestamp":1790975133243,"version":"4.1.0"},"reference-count":45,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T00:00:00Z","timestamp":1776988800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"China Agriculture Research System of MOF and MARA","award":["CARS-35"],"award-info":[{"award-number":["CARS-35"]}]},{"name":"Major Special Project for the Development of Agricultural Microbial Industry in Hubei Province","award":["NYWSWZX2025-2027-03"],"award-info":[{"award-number":["NYWSWZX2025-2027-03"]}]},{"name":"Hubei Province Postdoctoral Unveiling and Tackling Key Problems Program","award":["707124931"],"award-info":[{"award-number":["707124931"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vaccines"],"abstract":"<jats:p>Background\/Objectives: Extraintestinal pathogenic Escherichia coli (ExPEC) is a major pathogen that causes septicemia, meningitis, and polyserositis in pigs. The increasing prevalence of antimicrobial resistance and the diverse serotypes of ExPEC highlight the urgent need for broadly protective vaccines. Methods and Results: In this study, an OmpC epitope vaccine based on the T4 phage display system was developed and evaluated. Two B-cell epitopes (OmpC-1 and OmpC-2) were identified by bioinformatic analysis and displayed on recombinant T4 phages. Immunization induced strong antigen-specific IgG responses, with the OmpC-1-T4 group showing significantly higher antibody titers than the OmpC protein group. In the O11 serotype PCN033 challenge model, survival rates reached 100% in the OmpC-1-T4 group, 60% in the OmpC-2-T4 group, and approximately 80% in the OmpC protein group. In the O18 serotype 2103 challenge model, both recombinant phage groups had survival rates of approximately 60%, whereas all the mice in the OmpC protein group died within three days. OmpC-1-T4 immunization also significantly reduced bacterial loads in lung and brain tissues after PCN033 infection and decreased TNF-\u03b1 and IL-6 expression in lung tissues, accompanied by reduced inflammatory infiltration and tissue damage. Conclusions: Overall, the T4 phage-displayed OmpC epitope vaccine induced strong humoral immunity and provided protection against different ExPEC serotypes. Among the candidates, OmpC-1-T4 showed superior immune protection, bacterial clearance, and inflammation control, supporting its potential as a vaccine candidate against porcine ExPEC infection.<\/jats:p>","DOI":"10.3390\/vaccines14050383","type":"journal-article","created":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T14:28:23Z","timestamp":1777040903000},"page":"383","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Immune Protection Effect of an OmpC-Recombinant T4 Bacteriophage Vaccine Against Infection Caused by Extraintestinal Pathogenic Escherichia coli in Mice"],"prefix":"10.3390","volume":"14","author":[{"given":"Xin","family":"Zong","sequence":"first","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiting","family":"Ni","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guosheng","family":"Chen","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaodan","family":"Li","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaqi","family":"Liu","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7418-854X","authenticated-orcid":false,"given":"Ze","family":"Tong","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhengnan","family":"Yuan","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shiyuan","family":"Jiang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Huanchun","family":"Chen","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Hubei Hongshan Laboratory, Wuhan 430070, China"},{"name":"Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, China"},{"name":"Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chen","family":"Tan","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"},{"name":"Hubei Hongshan Laboratory, Wuhan 430070, China"},{"name":"Frontiers Science Center for Animal Breeding and Sustainable Production, Wuhan 430070, China"},{"name":"Key Laboratory of Preventive Veterinary Medicine in Hubei Province, Wuhan 430070, China"},{"name":"The Cooperative Innovation Center for Sustainable Pig Production, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-0761-8511","authenticated-orcid":false,"given":"Chenchen","family":"Wang","sequence":"additional","affiliation":[{"name":"National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Hussain, A., Najeeb, A., and Ali, S.A. (2025). Antimicrobial Resistance: A Modern Plague. AI-Driven Breakthroughs in Antimicrobial Resistance, IGI Global Scientific Publishing.","DOI":"10.4018\/979-8-3693-7550-1.ch002"},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Salam, M.A., Al-Amin, M.Y., Salam, M.T., Pawar, J.S., Akhter, N., Rabaan, A.A., and Alqumber, M.A. (2023). Antimicrobial resistance: A growing serious threat for global public health. Healthcare, 13.","DOI":"10.20944\/preprints202305.0555.v1"},{"key":"ref_3","first-page":"159","article-title":"Bacterial pathogens of pigs with particular reference to Escherichia coli: A systematic review and meta-analysis","volume":"9","author":"Abubakar","year":"2017","journal-title":"J. Vet. Med. Anim. Health"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Sora, V.M., Meroni, G., Martino, P.A., Soggiu, A., Bonizzi, L., and Zecconi, A. (2021). Extraintestinal pathogenic Escherichia coli: Virulence factors and antibiotic resistance. Pathogens, 10.","DOI":"10.3390\/pathogens10111355"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1186\/s44149-025-00158-x","article-title":"Unraveling the microbial contamination characteristics of pork farms and disinfection efficacy assessment via high-throughput sequencing","volume":"5","author":"Liu","year":"2025","journal-title":"Anim. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5811","DOI":"10.1038\/s41467-024-50268-2","article-title":"Population structure and antibiotic resistance of swine extraintestinal pathogenic Escherichia coli from China","volume":"15","author":"Li","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e00135-18","DOI":"10.1128\/CMR.00135-18","article-title":"Global extraintestinal pathogenic Escherichia coli (ExPEC) lineages","volume":"32","author":"Manges","year":"2019","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Hao, L., Huang, W., Guo, Y., Liu, X., Wu, J., Zhu, L., Pan, C., and Wang, H. (2025). A bioconjugate vaccine against extra-intestinal pathogenic Escherichia coli (ExPEC). Vaccines, 13.","DOI":"10.3390\/vaccines13040362"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1038\/nm.4465","article-title":"The role of vaccines in preventing bacterial antimicrobial resistance","volume":"24","author":"Jansen","year":"2018","journal-title":"Nat. Med."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"388","DOI":"10.1093\/femsre\/fuy011","article-title":"Potential targets for next generation antimicrobial glycoconjugate vaccines","volume":"42","author":"Micoli","year":"2018","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1038\/s41541-017-0006-8","article-title":"Escherichia coli-derived virus-like particles in vaccine development","volume":"2","author":"Huang","year":"2017","journal-title":"NPJ Vaccines"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"421","DOI":"10.1007\/s10719-021-09985-9","article-title":"The development and characterization of an E. coli O25B bioconjugate vaccine","volume":"38","author":"Kowarik","year":"2021","journal-title":"Glycoconj. J."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2359691","DOI":"10.1080\/19490976.2024.2359691","article-title":"Vaccines against extraintestinal pathogenic Escherichia coli (ExPEC): Progress and challenges","volume":"16","author":"Qiu","year":"2024","journal-title":"Gut Microbes"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Zhou, G., Wang, Q., Wang, Y., Wen, X., Peng, H., Peng, R., Shi, Q., Xie, X., and Li, L. (2023). Outer membrane porins contribute to antimicrobial resistance in gram-negative bacteria. Microorganisms, 11.","DOI":"10.3390\/microorganisms11071690"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1016\/S1286-4579(02)01545-9","article-title":"Outer membrane proteins: Key players for bacterial adaptation in host niches","volume":"4","author":"Lin","year":"2002","journal-title":"Microbes Infect."},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Masi, M., Winterhalter, M., and Pag\u00e8s, J.-M. (2019). Outer membrane porins. Bacterial Cell Walls and Membranes, Springer.","DOI":"10.1007\/978-3-030-18768-2_4"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2449722","DOI":"10.1080\/21505594.2025.2449722","article-title":"Klebsiella pneumoniae derived outer membrane vesicles mediated bacterial virulence, antibiotic resistance, host immune responses and clinical applications","volume":"16","author":"Li","year":"2025","journal-title":"Virulence"},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Elshobary, M.E., Badawy, N.K., Ashraf, Y., Zatioun, A.A., Masriya, H.H., Ammar, M.M., Mohamed, N.A., Mourad, S., and Assy, A.M. (2025). Combating antibiotic resistance: Mechanisms, multidrug-resistant pathogens, and novel therapeutic approaches: An updated review. Pharmaceuticals, 18.","DOI":"10.3390\/ph18030402"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"6819","DOI":"10.1007\/s00253-021-11511-8","article-title":"Identification of a specific surface epitope of OmpC for Escherichia coli O157: H7 with protein topology facilitated affinity mass spectrometry","volume":"105","author":"Wang","year":"2021","journal-title":"Appl. Microbiol. Biot."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Igomu, E.E., Mamman, P.H., Adamu, J., Muhammad, M., Woziri, A.O., Sugun, M.Y., Benshak, J.A., Anyika, K.C., Sam-Gyang, R., and Ehizibolo, D.O. (2025). Immunoinformatics design of a novel multiepitope vaccine candidate against non-typhoidal salmonellosis caused by Salmonella Kentucky using outer membrane proteins A, C, and F. PLoS ONE, 20.","DOI":"10.1371\/journal.pone.0306200"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kwang, J., and He, F. (2013). Microbes and livestock. Microbial Biotechnology: Principles and Applications, World Scientific.","DOI":"10.1142\/9789814366830_0014"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1048","DOI":"10.1073\/pnas.81.4.1048","article-title":"Amino acid sequence homology among the major outer membrane proteins of Escherichia coli","volume":"81","author":"Nikaido","year":"1984","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1194","DOI":"10.1016\/j.procbio.2015.05.001","article-title":"Paving the way to construct a new vaccine against Escherichia coli from its recombinant outer membrane protein C via a murine model","volume":"50","author":"Wang","year":"2015","journal-title":"Process Biochem."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"849","DOI":"10.1016\/j.biotechadv.2010.07.004","article-title":"Phage display: Concept, innovations, applications and future","volume":"28","author":"Pande","year":"2010","journal-title":"Biotechnol. Adv."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"eesp\u20130004\u20132025","DOI":"10.1128\/ecosalplus.esp-0004-2025","article-title":"Bacteriophage T4 genome packaging: Mechanism and application","volume":"13","author":"Rao","year":"2025","journal-title":"EcoSal Plus"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1007\/s00203-014-0989-8","article-title":"T4 bacteriophage as a phage display platform","volume":"196","author":"Gamkrelidze","year":"2014","journal-title":"Arch. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"12551","DOI":"10.1128\/JVI.02043-14","article-title":"Immunogenicity studies of proteins forming the T4 phage head surface","volume":"88","author":"Miernikiewicz","year":"2014","journal-title":"J. Virol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"104","DOI":"10.1111\/1574-6968.12013","article-title":"Immunogenic characterization of outer membrane porins OmpC and OmpF of porcine extraintestinal pathogenic Escherichia coli","volume":"337","author":"Liu","year":"2012","journal-title":"FEMS Microbiol. Lett."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"312","DOI":"10.2174\/1874609813666200929113341","article-title":"Designing an outer membrane protein (Omp-W) based vaccine for immunization against Vibrio and Salmonella: An in silico approach","volume":"14","author":"Tehrani","year":"2020","journal-title":"Recent Pat. Biotechnol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"721","DOI":"10.2217\/fmb-2020-0232","article-title":"Evaluation of the immunoprotective effects of IF-2 GTPase and SSU05-1022 as a candidate for a Streptococcus suis subunit vaccine","volume":"16","author":"Chen","year":"2021","journal-title":"Future Microbiol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e3573","DOI":"10.21769\/BioProtoc.3573","article-title":"Preparation of a Bacteriophage T4-based Prokaryotic-eukaryotic Hybrid Viral Vector for Delivery of Large Cargos of Genes and Proteins into Human Cells","volume":"10","author":"Zhu","year":"2020","journal-title":"Bio Protoc."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1007\/978-1-0716-1884-4_10","article-title":"CRISPR Engineering of Bacteriophage T4 to Design Vaccines Against SARS-CoV-2 and Emerging Pathogens","volume":"2410","author":"Zhu","year":"2022","journal-title":"Methods Mol. Biol. (Clifton N.J.)"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"eabh1547","DOI":"10.1126\/sciadv.abh1547","article-title":"A universal bacteriophage T4 nanoparticle platform to design multiplex SARS-CoV-2 vaccine candidates by CRISPR engineering","volume":"7","author":"Zhu","year":"2021","journal-title":"Sci. Adv."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2639","DOI":"10.1021\/acssynbio.1c00251","article-title":"Engineering T4 Bacteriophage for In Vivo Display by Type V CRISPR-Cas Genome Editing","volume":"10","author":"Dong","year":"2021","journal-title":"ACS Synth. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Tao, P., Li, Q., Shivachandra, S.B., and Rao, V.B. (2017). Bacteriophage T4 as a nanoparticle platform to display and deliver pathogen antigens: Construction of an effective anthrax vaccine. Recombinant Virus Vaccines: Methods and Protocols, Springer.","DOI":"10.1007\/978-1-4939-6869-5_15"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"33134","DOI":"10.1021\/acsnano.5c04881","article-title":"Self-Assembled Nanoparticle Vaccines Elicit Robust Protective Immune Responses against Type O Foot-and-Mouth Disease Virus Infection","volume":"19","author":"Cao","year":"2025","journal-title":"Acs Nano"},{"key":"ref_37","first-page":"162","article-title":"Evaluation of recombinant outer membrane protein based vaccine against Salmonella Typhimurium in birds","volume":"41","author":"Prejit","year":"2013","journal-title":"Biol. J. Int. Assoc. Biol. Stand."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1080\/07391102.2000.10506664","article-title":"Homology model of surface antigen OmpC from Salmonella typhi and its functional implications","volume":"18","author":"Arockiasamy","year":"2000","journal-title":"J. Biomol. Struct. Dyn."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s44149-024-00112-3","article-title":"Characterization and genomics analysis of phage PGX1 against multidrug-resistant enterotoxigenic E. coli with in vivo and in vitro efficacy assessment","volume":"4","author":"Hu","year":"2024","journal-title":"Anim. Dis."},{"key":"ref_40","doi-asserted-by":"crossref","unstructured":"Liu, S., Lin, M., and Zhou, X. (2025). T4 Phage Displaying Dual Antigen Clusters Against H3N2 Influenza Virus Infection. Vaccines, 13.","DOI":"10.3390\/vaccines13010070"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1146\/annurev-virology-111821-111145","article-title":"Bacteriophage T4 as a Protein-Based, Adjuvant- and Needle-Free, Mucosal Pandemic Vaccine Design Platform","volume":"11","author":"Zhu","year":"2024","journal-title":"Annu. Rev. Virol."},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Pokharel, P., Dhakal, S., and Dozois, C.M. (2023). The Diversity of Escherichia coli Pathotypes and Vaccination Strategies against This Versatile Bacterial Pathogen. Microorganisms, 11.","DOI":"10.3390\/microorganisms11020344"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1089\/fpd.2007.0087","article-title":"Extraintestinal pathogenic Escherichia coli","volume":"4","author":"Smith","year":"2007","journal-title":"Foodborne Pathog. Dis."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2255","DOI":"10.1056\/NEJMra2026131","article-title":"Cytokine Storm","volume":"383","year":"2020","journal-title":"N. Engl. J. Med."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"401","DOI":"10.1086\/589862","article-title":"Correlates of Vaccine-Induced Immunity","volume":"47","author":"Plotkin","year":"2008","journal-title":"Clin. Infect. Dis."}],"container-title":["Vaccines"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/5\/383\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,24]],"date-time":"2026-04-24T14:42:30Z","timestamp":1777041750000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-393X\/14\/5\/383"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,24]]},"references-count":45,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2026,5]]}},"alternative-id":["vaccines14050383"],"URL":"https:\/\/doi.org\/10.3390\/vaccines14050383","relation":{},"ISSN":["2076-393X"],"issn-type":[{"value":"2076-393X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,24]]}}}