{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T00:29:09Z","timestamp":1789000149057,"version":"build-2803163510"},"reference-count":43,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T00:00:00Z","timestamp":1768953600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100006606","name":"Natural Science Foundation of Tianjin","doi-asserted-by":"publisher","award":["24JCQNJC00460"],"award-info":[{"award-number":["24JCQNJC00460"]}],"id":[{"id":"10.13039\/501100006606","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Public Health"],"abstract":"<jats:sec>\n                    <jats:title>Introduction<\/jats:title>\n                    <jats:p>\n                      Influenza A and\n                      <jats:italic>Mycoplasma pneumoniae<\/jats:italic>\n                      (\n                      <jats:italic>M. pneumoniae<\/jats:italic>\n                      ) are common respiratory pathogens, causing severe co-infections in clinical diseases. Current vaccines have failed to provide comprehensive protection against both of the pathogens, highlighting the requirement of integrated solution.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      In this study, a novel dual-target multi-epitope vaccine was developed based on the immunoinformatics method. Based on the consistent sequences, we used the HA and NA proteins of influenza A virus, and the p1, p65, HMW1-3 proteins of\n                      <jats:italic>M. pneumoniae<\/jats:italic>\n                      to determine the immunodominant T- and B-cell epitopes.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results and Discussion<\/jats:title>\n                    <jats:p>\n                      The designed vaccine included 21 linear B-cell epitopes, 34 CTL epitopes and 19 HTL epitopes from influenza A virus and\n                      <jats:italic>M. pneumoniae<\/jats:italic>\n                      . The selection was based on antigenicity, immunogenicity, and alignment with previously validated epitopes. Structural and physicochemical assessments indicated that the vaccine had high stability, solubility, and minimal allergy risk. Molecular docking with TLR3 and TLR4 receptors revealed strong binding, suggesting robust immune activation. Population coverage analysis showed the vaccine could cover 97.07% of the global population, with high efficacy across key regions such as North America and East Asia. In this study, we provided an immunoinformatics approach for vaccine design.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.3389\/fpubh.2026.1671035","type":"journal-article","created":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T06:51:16Z","timestamp":1768978276000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":3,"title":["Designing a broad-spectrum multi-epitope vaccine against influenza A and Mycoplasma pneumoniae: an immunoinformatics approach"],"prefix":"10.3389","volume":"14","author":[{"given":"Zhengyu","family":"Yang","sequence":"first","affiliation":[{"name":"Intellectual Property Research Institute, Xiamen University","place":["Xiamen, Fujian, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Li","sequence":"additional","affiliation":[{"name":"National Engineering Research Center for Beijing Biochip Technology (CapitalBio)","place":["Beijing, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lingling","family":"Chen","sequence":"additional","affiliation":[{"name":"School of Basic Medical Sciences, Tianjin Medical University","place":["Tianjin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shulei","family":"Jia","sequence":"additional","affiliation":[{"name":"School of Basic Medical Sciences, Tianjin Medical University","place":["Tianjin, China"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2026,1,21]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","first-page":"91","DOI":"10.1016\/j.molimm.2023.07.011","article-title":"\u0394A146Ply-HA stem protein immunization protects mice against influenza a virus infection and co-infection with Streptococcus pneumoniae","volume":"161","author":"Li","year":"2023","journal-title":"Mol Immunol"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"e32245","DOI":"10.1097\/md.0000000000032245","article-title":"Clinical features and outcome of influenza pneumonia in critically-ill immunocompromised patients","volume":"101","author":"Raymond","year":"2022","journal-title":"Medicine"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"182","DOI":"10.1186\/s12931-025-03262-1","article-title":"Development and validation of an early diagnosis model for severe Mycoplasma pneumonia in children based on interpretable machine learning","volume":"26","author":"Xie","year":"2025","journal-title":"Respir Res"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"329","DOI":"10.1007\/s40121-023-00914-x","article-title":"Clinical characteristics of lung consolidation with Mycoplasma pneumoniae pneumonia and risk factors for Mycoplasma pneumoniae necrotizing pneumonia in children","volume":"13","author":"Zhang","year":"2024","journal-title":"Infect Dis Ther"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"1437","DOI":"10.1007\/s10096-024-04854-3","article-title":"Risk factors for pneumonia among children with coinfection of influenza a virus and Mycoplasma pneumoniae","volume":"43","author":"Fu","year":"2024","journal-title":"Eur J Clin Microbiol Infect Dis"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"585","DOI":"10.1186\/s12890-024-03380-4","article-title":"Significance of respiratory virus coinfection in children with Mycoplasma pneumoniae pneumonia","volume":"24","author":"Yu","year":"2024","journal-title":"BMC Pulm Med"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"212","DOI":"10.1186\/s12890-022-02005-y","article-title":"Clinical significance of respiratory virus coinfection in children with Mycoplasma pneumoniae pneumonia","volume":"22","author":"Choo","year":"2022","journal-title":"BMC Pulm Med"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1007\/s12275-023-00076-y","article-title":"NEDD4 regulated pyroptosis occurred from co-infection between influenza a virus and Streptococcus pneumoniae","volume":"61","author":"You","year":"2023","journal-title":"J Microbiol"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"e04489-22","DOI":"10.1128\/spectrum.04489-22","article-title":"Vimentin is an attachment receptor for Mycoplasma pneumoniae p1 protein","volume":"11","author":"Peng","year":"2023","journal-title":"Microbiol Spectr"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"105647","DOI":"10.1016\/j.micpath.2022.105647","article-title":"Cell damage and neutrophils promote the infection of Mycoplasma pneumoniae and inflammatory response","volume":"169","author":"Zhang","year":"2022","journal-title":"Microb Pathog"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"44","DOI":"10.1016\/j.jdsr.2023.12.006","article-title":"Respiratory tract barrier dysfunction in viral-bacterial co-infection cases","volume":"60","author":"Sumitomo","year":"2024","journal-title":"Japan Dental Sci Rev"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"244","DOI":"10.1186\/1743-422X-10-244","article-title":"Cross-reactive human B cell and T cell epitopes between influenza a and B viruses","volume":"10","author":"Terajima","year":"2013","journal-title":"Virol J"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"162","DOI":"10.1186\/s12985-024-02429-4","article-title":"Application of peripheral blood routine parameters in the diagnosis of influenza and Mycoplasma pneumoniae","volume":"21","author":"Chen","year":"2024","journal-title":"Virol J"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1186\/s13568-025-01829-1","article-title":"The value of common blood parameters for the differential diagnosis of respiratory tract infections in children","volume":"15","author":"Ma","year":"2025","journal-title":"AMB Express"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"105194","DOI":"10.1016\/j.compbiomed.2021.105194","article-title":"An immunoinformatics-based designed multi-epitope candidate vaccine (mpme-VAC\/STV-1) against Mycoplasma pneumoniae","volume":"142","author":"Vilela Rodrigues","year":"2022","journal-title":"Comput Biol Med"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"W449","DOI":"10.1093\/nar\/gkaa379","article-title":"NetMHCpan-4.1 and NetMHCIIpan-4.0: improved predictions of MHC antigen presentation by concurrent motif deconvolution and integration of MS MHC eluted ligand data","volume":"48","author":"Reynisson","year":"2020","journal-title":"Nucleic Acids Res"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"114599","DOI":"10.1016\/j.colsurfb.2025.114599","article-title":"Designing a T cell multi-epitope vaccine against hRSV with reverse vaccinology: an immunoinformatics approach","volume":"251","author":"Guan","year":"2025","journal-title":"Colloids Surf B Biointerfaces"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1186\/1471-2105-8-4","article-title":"VaxiJen: a server for prediction of protective antigens, tumour antigens and subunit vaccines","volume":"8","author":"Doytchinova","year":"2007","journal-title":"BMC Bioinform"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1007\/978-1-4939-2285-7_7","article-title":"Peptide toxicity prediction","volume":"1268","author":"Gupta","year":"2015","journal-title":"Methods Mol Biol"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"107388","DOI":"10.1016\/j.actatropica.2024.107388","article-title":"Rational design of a multi-epitope vaccine against heartland virus (HRTV) using immune-informatics, molecular docking and dynamics approaches","volume":"259","author":"Ahmed","year":"2024","journal-title":"Acta Trop"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"W271","DOI":"10.1093\/nar\/gku339","article-title":"iMODS: internal coordinates normal mode analysis server","volume":"42","author":"L\u00f3pez-Blanco","year":"2014","journal-title":"Nucleic Acids Res"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"1546254","DOI":"10.3389\/fimmu.2025.1546254","article-title":"Design of a peptide-based vaccine against human respiratory syncytial virus using a reverse vaccinology approach: evaluation of immunogenicity, antigenicity, allergenicity, and toxicity","volume":"16","author":"Alnajran","year":"2025","journal-title":"Front Immunol"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"871","DOI":"10.1016\/j.ijbiomac.2020.07.117","article-title":"Design of a multi-epitope vaccine against SARS-CoV-2 using immunoinformatics approach","volume":"164","author":"Sanami","year":"2020","journal-title":"Int J Biol Macromol"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"10976","DOI":"10.1073\/pnas.0505077102","article-title":"The molecular structure of the toll-like receptor 3 ligand-binding domain","volume":"102","author":"Bell","year":"2005","journal-title":"Proc Natl Acad Sci USA"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"1191","DOI":"10.1038\/nature07830","article-title":"The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex","volume":"458","author":"Park","year":"2009","journal-title":"Nature"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"885","DOI":"10.1016\/j.ijbiomac.2022.12.066","article-title":"Designing multi-epitope mRNA construct as a universal influenza vaccine candidate for future epidemic\/pandemic preparedness","volume":"226","author":"Rcheulishvili","year":"2023","journal-title":"Int J Biol Macromol"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"3883","DOI":"10.1016\/j.vaccine.2024.04.098","article-title":"Design of a novel multiepitope vaccine with CTLA-4 extracellular domain against Mycoplasma pneumoniae: a vaccine-immunoinformatics approach","volume":"42","author":"Pan","year":"2024","journal-title":"Vaccine"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"105355","DOI":"10.1016\/j.meegid.2022.105355","article-title":"Insight into the first multi-epitope-based peptide subunit vaccine against avian influenza a virus (H5N6): an immunoinformatics approach","volume":"104","author":"Mia","year":"2022","journal-title":"Infect Genet Evol"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"67","DOI":"10.1186\/s12985-024-02337-7","article-title":"A computational approach to design a multiepitope vaccine against H5N1 virus","volume":"21","author":"Dashti","year":"2024","journal-title":"Virol J"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1186\/s12859-023-05374-1","article-title":"Beyond the state of the art of reverse vaccinology: predicting vaccine efficacy with the universal immune system simulator for influenza","volume":"24","author":"Russo","year":"2023","journal-title":"BMC Bioinform"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"1047306","DOI":"10.3389\/fcimb.2022.1047306","article-title":"Evaluation of the consistence between the results of immunoinformatics predictions and real-world animal experiments of a new tuberculosis vaccine MP3RT","volume":"12","author":"Cheng","year":"2022","journal-title":"Front Cell Infect Microbiol"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1186\/s43141-023-00574-x","article-title":"Computational design and evaluation of mRNA-and protein-based conjugate vaccines for influenza a and SARS-CoV-2 viruses","volume":"21","author":"Elalouf","year":"2023","journal-title":"J Genet Eng Biotechnol"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"1249718","DOI":"10.3389\/fmicb.2023.1249718","article-title":"TLR agonists as vaccine adjuvants in the prevention of viral infections: an overview","volume":"14","author":"Kayesh","year":"2023","journal-title":"Front Microbiol"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"339","DOI":"10.1016\/j.coi.2009.05.003","article-title":"Immunology of TLR-independent vaccine adjuvants","volume":"21","author":"De Gregorio","year":"2009","journal-title":"Curr Opin Immunol"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"1494017","DOI":"10.3389\/fimmu.2024.1494017","article-title":"Toll-like receptor 4 pathway evolutionary trajectory and functional emergence","volume":"15","author":"Verma","year":"2025","journal-title":"Front Immunol"},{"key":"ref36","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1093\/protein\/4.2.155","article-title":"Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence","volume":"4","author":"Guruprasad","year":"1990","journal-title":"Protein Eng"},{"key":"ref37","doi-asserted-by":"publisher","first-page":"104795","DOI":"10.1016\/j.meegid.2021.104795","article-title":"Rational design of multimeric based subunit vaccine against Mycoplasma pneumonia: subtractive proteomics with immunoinformatics framework","volume":"91","author":"Mahmood","year":"2021","journal-title":"Infect Genet Evol"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"6059","DOI":"10.1007\/s11030-025-11124-7","article-title":"Designing a multi-epitope vaccine candidate against pandemic influenza a virus: an immunoinformatics and structural vaccinology approach","volume":"29","author":"Samantaray","year":"2025","journal-title":"Mol Divers"},{"key":"ref39","doi-asserted-by":"publisher","first-page":"103283","DOI":"10.1016\/j.sjbs.2022.103283","article-title":"In silico epitope-based vaccine design against influenza a neuraminidase protein: computational analysis established on B- and T-cell epitope predictions","volume":"29","author":"Almalki","year":"2022","journal-title":"Saudi J Biol Sci"},{"key":"ref40","doi-asserted-by":"publisher","first-page":"e70021","DOI":"10.1002\/iid3.70021","article-title":"Research of recombinant influenza a virus as a vector for Mycoplasma pneumoniae P1a and P30a","volume":"12","author":"Yu","year":"2024","journal-title":"Immun Inflamm Dis"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"64","DOI":"10.1038\/s41541-024-00857-5","article-title":"An intranasal combination vaccine induces systemic and mucosal immunity against COVID-19 and influenza","volume":"9","author":"Xing","year":"2024","journal-title":"NPJ Vaccines"},{"key":"ref42","doi-asserted-by":"publisher","first-page":"e19371","DOI":"10.2196\/19371","article-title":"Structural basis for designing multiepitope vaccines against COVID-19 infection: in silico vaccine design and validation","volume":"1","author":"Srivastava","year":"2020","journal-title":"JMIR Bioinform Biotechnol"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1016\/j.molimm.2025.03.019","article-title":"HLA class I and II allelic diversity among ecuadorian transplant candidates: a comprehensive retrospective analysis","volume":"182","author":"Velastegu\u00ed","year":"2025","journal-title":"Mol Immunol"}],"container-title":["Frontiers in Public Health"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpubh.2026.1671035\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,2,24]],"date-time":"2026-02-24T07:40:50Z","timestamp":1771918850000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fpubh.2026.1671035\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,21]]},"references-count":43,"alternative-id":["10.3389\/fpubh.2026.1671035"],"URL":"https:\/\/doi.org\/10.3389\/fpubh.2026.1671035","relation":{},"ISSN":["2296-2565"],"issn-type":[{"value":"2296-2565","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,21]]},"article-number":"1671035"}}