{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T12:11:29Z","timestamp":1790424689319,"version":"4.1.0"},"reference-count":36,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,7,28]],"date-time":"2020-07-28T00:00:00Z","timestamp":1595894400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,7,28]],"date-time":"2020-07-28T00:00:00Z","timestamp":1595894400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100006083","name":"Ministry of Defence","doi-asserted-by":"publisher","award":["FDM0961722"],"award-info":[{"award-number":["FDM0961722"]}],"id":[{"id":"10.13039\/501100006083","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Technology Development Program for Bio-industry","award":["1116043-1"],"award-info":[{"award-number":["1116043-1"]}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["BMC Vet Res"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>\n                      <jats:italic>Bacillus anthracis<\/jats:italic>\n                      is the causative agent of anthrax, a disease of both humans and various animal species, and can be used as a bioterror agent. Effective vaccines are available, but those could benefit from improvements, including increasing the immunity duration, reducing the shot frequency and adverse reactions. In addition, more sophisticated antigen delivery and potentiation systems are urgently required.\n                    <\/jats:p>\n                    <jats:p>\n                      The protective antigen (PA), one of three major virulence factors associated with anthrax was displayed on the surface of\n                      <jats:italic>Bacillus subtilis<\/jats:italic>\n                      spores, which is a vaccine production host and delivery vector with several advantages such as a low production cost, straightforward administration as it is safe for human consumption and the particulate adjuvanticity. Mice were immunized orally (PO), intranasally (IN), sublingually (SL) or intraperitoneally (IP) with the PA displaying probiotic spore vaccine. Clinical observation, serological analysis and challenge experiment were conducted to investigate the safety and efficacy of the vaccine.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>\n                      A\/J mice immunized with the PA spore vaccine via PO, IN, SL, and IP were observed to have increased levels of active antibody titer, isotype profiles and toxin neutralizing antibody in sera, and IgA in saliva. The immunized mice were demonstrated to raise protective immunity against the challenge with lethal\n                      <jats:italic>B. anthracis<\/jats:italic>\n                      spores.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>\n                      In this study, we developed a\n                      <jats:italic>B. subtilis<\/jats:italic>\n                      spore vaccine that displays the PA on its surface and showed that the PA-displaying spore vaccine was able to confer active immunity to a murine model based on the results of antibody isotype titration, mucosal antibody identification, and a lethal challenge experiment.\n                    <\/jats:p>\n                  <\/jats:sec>","DOI":"10.1186\/s12917-020-02468-3","type":"journal-article","created":{"date-parts":[[2020,7,28]],"date-time":"2020-07-28T09:04:34Z","timestamp":1595927074000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":28,"title":["Bacillus subtilis spore vaccines displaying protective antigen induce functional antibodies and protective potency"],"prefix":"10.1186","volume":"16","author":[{"given":"Yeonsu","family":"Oh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung Ae","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chang-Hwan","family":"Kim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Soo-Keun","family":"Choi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jae-Gu","family":"Pan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,7,28]]},"reference":[{"key":"2468_CR1","doi-asserted-by":"publisher","first-page":"647","DOI":"10.1146\/annurev.micro.55.1.647","volume":"55","author":"M Mock","year":"2001","unstructured":"Mock M, Fouet A. Anthrax. Annu Rev Microbiol. 2001;55:647\u201371.","journal-title":"Annu Rev Microbiol"},{"issue":"2","key":"2468_CR2","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1586\/14787210.7.2.219","volume":"7","author":"JN Tournier","year":"2009","unstructured":"Tournier JN, Ulrich RG, Quesnel-Hellmann A, Mohamadzadeh M, Stiles BG. Anthrax, toxins and vaccines: a 125-year journey targeting Bacillus anthracis. Expert Rev Anti-Infect Ther. 2009;7(2):219\u201336.","journal-title":"Expert Rev Anti-Infect Ther"},{"issue":"2","key":"2468_CR3","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1097\/00001432-200004000-00004","volume":"13","author":"PCB Turnbull","year":"2000","unstructured":"Turnbull PCB. Current status of immunization against anthrax: old vaccines may be here to stay for a while. Curr Opin Infect Dis. 2000;13(2):113\u201320.","journal-title":"Curr Opin Infect Dis"},{"issue":"3","key":"2468_CR4","doi-asserted-by":"publisher","first-page":"148","DOI":"10.2174\/187220811797579097","volume":"5","author":"M Kaur","year":"2011","unstructured":"Kaur M, Bhatnagar R. Recent progress in the development of anthrax vaccines. Recent Pat Biotechnol. 2011;5(3):148\u201359.","journal-title":"Recent Pat Biotechnol"},{"key":"2468_CR5","doi-asserted-by":"publisher","first-page":"1079","DOI":"10.1021\/bi015860m","volume":"41","author":"J Mogridge","year":"2002","unstructured":"Mogridge J, Cunningham K, Collier RJ. Stoichiometry of anthrax toxin complexes. Biochemistry. 2002;41:1079\u201382.","journal-title":"Biochemistry."},{"key":"2468_CR6","doi-asserted-by":"publisher","first-page":"813","DOI":"10.1016\/j.femsre.2004.12.001","volume":"29","author":"HA Hong","year":"2005","unstructured":"Hong HA, Duc LH, Cutting SM. The use of bacterial spore formers as probiotics. FEMS Microbiol Rev. 2005;29:813\u201335.","journal-title":"FEMS Microbiol Rev"},{"key":"2468_CR7","doi-asserted-by":"publisher","first-page":"4671","DOI":"10.1016\/j.vaccine.2007.04.021","volume":"25","author":"JD Paccez","year":"2007","unstructured":"Paccez JD, Nguyen HD, Luiz WB, Ferreira RCC, Sbrogio-Almeida ME, Ferreira LCS. Evaluation of different promoter sequences and antigen sorting signals on the immunogenicity of Bacillus subtillis vaccine vehicles. Vaccine. 2007;25:4671\u201380.","journal-title":"Vaccine"},{"issue":"7","key":"2468_CR8","doi-asserted-by":"publisher","first-page":"979","DOI":"10.4161\/hv.20694","volume":"8","author":"H Amuguni","year":"2012","unstructured":"Amuguni H, Tzipori S. Bacillus subtilis: a temperature resistant and needle free delivery system of immunogens. Hum Vaccin Immunother. 2012;8(7):979\u201386.","journal-title":"Hum Vaccin Immunother"},{"issue":"12","key":"2468_CR9","doi-asserted-by":"publisher","first-page":"610","DOI":"10.1016\/j.tibtech.2012.09.005","volume":"30","author":"JG Pan","year":"2012","unstructured":"Pan JG, Kim EJ, Yun CH. Bacillus spore display. Trends Biotechnol. 2012;30(12):610\u20132.","journal-title":"Trends Biotechnol"},{"issue":"6","key":"2468_CR10","doi-asserted-by":"publisher","first-page":"1538","DOI":"10.1002\/eji.200636875","volume":"37","author":"AG Barnes","year":"2007","unstructured":"Barnes AG, Cerovic V, Hobson PS, Klavinskis LS. Bacillus subtilis spores: a novel microparticle adjuvant which can instruct a balanced Th1 and Th2 immune response to specific antigen. Eur J Immunol. 2007;37(6):1538\u201347.","journal-title":"Eur J Immunol"},{"issue":"11","key":"2468_CR11","doi-asserted-by":"publisher","first-page":"1975","DOI":"10.1038\/15058","volume":"17","author":"M Singh","year":"1999","unstructured":"Singh M, O\u2019Hagan D. Advances in vaccine adjuvants. Nat Biotechnol. 1999;17(11):1975\u201381.","journal-title":"Nat Biotechnol"},{"issue":"5","key":"2468_CR12","doi-asserted-by":"publisher","first-page":"2810","DOI":"10.1128\/IAI.71.5.2810-2818.2003","volume":"71","author":"H Duc le","year":"2003","unstructured":"Duc le H, Hong HA, Fairweather N, Ricca E, Cutting SM. Bacterial spores as vaccine vehicles. Infect Immun. 2003;71(5):2810\u20138.","journal-title":"Infect Immun"},{"issue":"2","key":"2468_CR13","doi-asserted-by":"publisher","first-page":"346","DOI":"10.1016\/j.vaccine.2006.07.037","volume":"25","author":"H Duc le","year":"2007","unstructured":"Duc le H, Hong HA, Atkins HS, Flick-Smith HC, Durrani Z, Rijpkema S, Titball RW, Cutting SM. Immunization against anthrax using Bacillus subtilis spores expressing the anthrax protective antigen. Vaccine. 2007;25(2):346\u201355.","journal-title":"Vaccine"},{"issue":"5","key":"2468_CR14","doi-asserted-by":"publisher","first-page":"788","DOI":"10.1016\/j.vaccine.2006.09.013","volume":"25","author":"EM Mauriello","year":"2007","unstructured":"Mauriello EM, Cangiano G, Maurano F, Saggese V, De Felice M, Rossi M, Ricca E. Germination-independent induction of cellular immune response by Bacillus subtilis spores displaying the C fragment of the tetanus toxin. Vaccine. 2007;25(5):788\u201393.","journal-title":"Vaccine"},{"issue":"2","key":"2468_CR15","doi-asserted-by":"publisher","first-page":"209","DOI":"10.1111\/1574-6968.12558","volume":"358","author":"JG Pan","year":"2014","unstructured":"Pan JG, Choi SK, Jung HC, Kim EJ. Display of native proteins on Bacillus subtilis spores. FEMS Microbiol Lett. 2014;358(2):209\u201317.","journal-title":"FEMS Microbiol Lett"},{"issue":"11","key":"2468_CR16","doi-asserted-by":"publisher","first-page":"1647","DOI":"10.1128\/CVI.00135-10","volume":"17","author":"S Lee","year":"2010","unstructured":"Lee S, Belitsky BR, Brinker JP, Kerstein KO, Brown DW, Clements JD, Keusch GT, Tzipori S, Sonenshein AL, Herrmann JE. Development of a Bacillus subtilis-based rotavirus vaccine. Clin Vaccine Immunol. 2010;17(11):1647\u201355.","journal-title":"Clin Vaccine Immunol"},{"key":"2468_CR17","volume-title":"Gram-positive bacteria. Vaccine vehicles for mucosal immunization","year":"1997","unstructured":"Pozzi G, Wells JM, editors. Gram-positive bacteria. Vaccine vehicles for mucosal immunization. Berlin: Springer-Verlag; 1997."},{"issue":"2","key":"2468_CR18","doi-asserted-by":"publisher","first-page":"129","DOI":"10.1016\/S0168-1656(02)00043-3","volume":"96","author":"P Samuelson","year":"2002","unstructured":"Samuelson P, Gunneriusson E, Nygren PA, St\u00e5hl S. Display of proteins on bacteria. J Biotechnol. 2002;96(2):129\u201354.","journal-title":"J Biotechnol"},{"issue":"1","key":"2468_CR19","doi-asserted-by":"publisher","first-page":"29","DOI":"10.1038\/nbt0197-29","volume":"15","author":"G Georgiou","year":"1997","unstructured":"Georgiou G, Stathopoulos C, Daugherty PS, Nayak AR, Iverson BL, Curtiss R 3rd. Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines. Nat Biotechnol. 1997;15(1):29\u201334.","journal-title":"Nat Biotechnol"},{"issue":"2","key":"2468_CR20","doi-asserted-by":"publisher","first-page":"79","DOI":"10.1016\/j.tibtech.2010.11.003","volume":"29","author":"E van Bloois","year":"2011","unstructured":"van Bloois E, Winter RT, Kolmar H, Fraaije MW. Decorating microbes: surface display of proteins on Escherichia coli. Trends Biotechnol. 2011;29(2):79\u201386.","journal-title":"Trends Biotechnol"},{"key":"2468_CR21","first-page":"115","volume":"13","author":"E Ricca","year":"2014","unstructured":"Ricca E, Baccigalupi L, Cangiano G, De Felice M, Isticato R. Mucosal vaccine delivery by non-recombinant spores of Bacillus subtilis. Microb Cell Factories. 2014;13:115.","journal-title":"Microb Cell Factories"},{"issue":"2","key":"2468_CR22","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1111\/j.1574-695X.2008.00415.x","volume":"53","author":"JM Huang","year":"2008","unstructured":"Huang JM, La Ragione RM, Nunez A, Cutting SM. Immunostimulatory activity of Bacillus spores. FEMS Immunol Med Microbiol. 2008;53(2):195\u2013203.","journal-title":"FEMS Immunol Med Microbiol"},{"issue":"22","key":"2468_CR23","doi-asserted-by":"publisher","first-page":"3266","DOI":"10.1016\/j.vaccine.2012.03.016","volume":"30","author":"M Song","year":"2012","unstructured":"Song M, Hong HA, Huang JM, Colenutt C, Khang DD, Nguyen TV, Park SM, Shim BS, Song HH, Cheon IS, Jang JE, Choi JA, Choi YK, Stadler K, Cutting SM. Killed Bacillus subtilis spores as a mucosal adjuvant for an H5N1 vaccine. Vaccine. 2012;30(22):3266\u201377.","journal-title":"Vaccine"},{"key":"2468_CR24","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1111\/j.1749-6632.1983.tb26868.x","volume":"409","author":"SJ Challacombe","year":"1983","unstructured":"Challacombe SJ. Salivary antibodies and systemic tolerance in mice after oral immunization with bacterial antigens. Ann N Y Acad Sci. 1983;409:177\u201392.","journal-title":"Ann N Y Acad Sci"},{"issue":"11","key":"2468_CR25","doi-asserted-by":"publisher","first-page":"6883","DOI":"10.4049\/jimmunol.0901466","volume":"183","author":"IM Belyakov","year":"2009","unstructured":"Belyakov IM, Ahlers JD. What role does the route of immunization play in the generation of protective immunity against mucosal pathogens? J Immunol. 2009;183(11):6883\u201392.","journal-title":"J Immunol"},{"key":"2468_CR26","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1016\/0022-1759(90)90151-K","volume":"126","author":"BJ Weiblen","year":"1990","unstructured":"Weiblen BJ, Schumacher RT, Hoff R. Detection of IgM and IgA HIV antibodies after removal of IgG with recombinant protein G. J Immunol Methods. 1990;126:199\u2013204.","journal-title":"J Immunol Methods"},{"issue":"8","key":"2468_CR27","doi-asserted-by":"publisher","first-page":"4020","DOI":"10.1128\/IAI.00070-07","volume":"75","author":"AU Bielinska","year":"2007","unstructured":"Bielinska AU, Janczak KW, Landers JJ, Makidon P, Sower LE, Peterson JW, Baker JR. Mucosal immunization with a novel nanoemulsion-based recombinant anthrax protective antigen vaccine protects against Bacillus anthracis spore challenge. Infect Immun. 2007;75(8):4020\u20139.","journal-title":"Infect Immun"},{"key":"2468_CR28","doi-asserted-by":"publisher","first-page":"6658","DOI":"10.1016\/j.vaccine.2010.08.016","volume":"28","author":"S Lee","year":"2010","unstructured":"Lee S, Belitsky BR, Brown DW, Brinker JP, Kerstein KO, Herrmann JE, Keusch GT, Sonenshein AL, Tzipori S. Efficacy, heat stability and safety of intranasally administered Bacillus subtilis spore or vegetative cell vaccines expressing tetanus toxin fragment C. Vaccine. 2010;28:6658\u201365.","journal-title":"Vaccine."},{"key":"2468_CR29","doi-asserted-by":"publisher","first-page":"653","DOI":"10.1038\/nbt0797-653","volume":"15","author":"K Robinson","year":"1997","unstructured":"Robinson K, Chamberlain LM, Schofield KM, Wells JM, Le Page RWF. Oral vaccination of mice against tetanus with recombinant Lactococcus lactis. Nat Biotech. 1997;15:653\u20137.","journal-title":"Nat Biotech"},{"key":"2468_CR30","doi-asserted-by":"publisher","first-page":"442","DOI":"10.1128\/JB.160.1.442-444.1984","volume":"160","author":"F Kawamura","year":"1984","unstructured":"Kawamura F, Doi RH. Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases. J Bacteriol. 1984;160:442\u20134.","journal-title":"J Bacteriol"},{"key":"2468_CR31","first-page":"e01380","volume":"7","author":"H Jeong","year":"2018","unstructured":"Jeong H, Jeong DE, Park SH, Kim SJ, Choi SK. Complete genome sequence of Bacillus subtilis strain WB800N, an extracellular protease-deficient derivative of strain 168. Microbiol Resourc Announc. 2018;7:e01380\u201318.","journal-title":"Microbiol Resourc Announc"},{"key":"2468_CR32","doi-asserted-by":"publisher","first-page":"50","DOI":"10.1016\/j.jbiotec.2017.12.007","volume":"266","author":"DE Jeong","year":"2018","unstructured":"Jeong DE, So Y, Park SY, Park SH, Choi SK. Random knock-in expression system for high yield production of heterologous protein in Bacillus subtilis. J Biotechnol. 2018;266:50\u20138.","journal-title":"J Biotechnol"},{"issue":"3","key":"2468_CR33","doi-asserted-by":"crossref","first-page":"981","DOI":"10.4049\/jimmunol.149.3.981","volume":"149","author":"S Tamura","year":"1992","unstructured":"Tamura S, Ito Y, Asanuma H, Hirabayashi Y, Suzuki Y, Nagamine T, Aizawa C, Kurata T. Cross-protection against influenza virus infection afforded by trivalent inactivated vaccines inoculated intranasally with cholera toxin B subunit. J Immunol. 1992;149(3):981\u20138.","journal-title":"J Immunol"},{"issue":"5","key":"2468_CR34","doi-asserted-by":"publisher","first-page":"1644","DOI":"10.1073\/pnas.0708684105","volume":"105","author":"JH Song","year":"2008","unstructured":"Song JH, Nguyen HH, Cuburu N, Horimoto T, Ko SY, Park SH, Czerkinsky C, Kweon MN. Sublingual vaccination with influenza virus protects mice against lethal viral infection. PNAS. 2008;105(5):1644\u20139.","journal-title":"PNAS."},{"key":"2468_CR35","doi-asserted-by":"crossref","first-page":"3616","DOI":"10.4049\/jimmunol.161.7.3616","volume":"161","author":"AL Mora","year":"1998","unstructured":"Mora AL, Tam JP. Controlled lipidation and encapsulation of peptides as a useful approach to mucosal immunizations. J Immunol. 1998;161:3616\u201323.","journal-title":"J Immunol"},{"key":"2468_CR36","doi-asserted-by":"publisher","first-page":"17","DOI":"10.1016\/j.biologicals.2003.09.003","volume":"32","author":"D Hering","year":"2004","unstructured":"Hering D, Thompson W, Hewetson J, Little S, Norris S, Pace-Templeton J. Validation of the anthrax lethal toxin neutralization assay. Biologicals. 2004;32:17\u201327.","journal-title":"Biologicals."}],"container-title":["BMC Veterinary Research"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12917-020-02468-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s12917-020-02468-3\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s12917-020-02468-3.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,4]],"date-time":"2023-10-04T19:24:10Z","timestamp":1696447450000},"score":1,"resource":{"primary":{"URL":"https:\/\/bmcvetres.biomedcentral.com\/articles\/10.1186\/s12917-020-02468-3"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,28]]},"references-count":36,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["2468"],"URL":"https:\/\/doi.org\/10.1186\/s12917-020-02468-3","relation":{"has-preprint":[{"id-type":"doi","id":"10.21203\/rs.3.rs-28219\/v1","asserted-by":"object"},{"id-type":"doi","id":"10.21203\/rs.3.rs-28219\/v2","asserted-by":"object"},{"id-type":"doi","id":"10.21203\/rs.3.rs-28219\/v3","asserted-by":"object"}]},"ISSN":["1746-6148"],"issn-type":[{"value":"1746-6148","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,28]]},"assertion":[{"value":"23 October 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 July 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"28 July 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"All experiments were performed in accordance with the Institutional Guidelines for the Care and Use of Laboratory Animals in Research and the approval of the Animal Care and Use Committee of the Korean Research Institute of Bioscience and Biotechnology (KRIBB-AEC-13069).","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"Not applicable.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"No conflict of interest declared.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"259"}}