{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T14:22:18Z","timestamp":1782915738823,"version":"3.54.5"},"reference-count":53,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2021,5,18]],"date-time":"2021-05-18T00:00:00Z","timestamp":1621296000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000060","name":"National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["U01AI124289"],"award-info":[{"award-number":["U01AI124289"]}],"id":[{"id":"10.13039\/100000060","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathogens"],"abstract":"<jats:p>Non-typhoidal salmonellosis, caused by Salmonella enterica serovar Typhimurium is a common fecal-oral disease characterized by mild gastrointestinal distress resulting in diarrhea, chills, fever, abdominal cramps, head and body aches, nausea, and vomiting. Increasing incidences of antibiotic resistant invasive non-typhoidal Salmonella infections makes this a global threat requiring novel treatment strategies including next-generation vaccines. The goal of the current study was to formulate a novel vaccine platform against Salmonella infection that could be delivered orally. To accomplish this, we created a Salmonella-specific vaccine adjuvanted with Burkholderia pseudomallei outer membrane vesicles (OMVs). We show that adding OMVs to a heat-killed oral Salmonella vaccine (HKST + OMVs) protects against a lethal, oral challenge with Salmonella. Further, we show that opsonizing anti-Salmonella antibodies are induced in response to immunization and that CD4 T cells and B cells can be induced when OMVs are used as the oral adjuvant. This study represents a novel oral vaccine approach to combatting the increasing problem of invasive Salmonella infections.<\/jats:p>","DOI":"10.3390\/pathogens10050616","type":"journal-article","created":{"date-parts":[[2021,5,18]],"date-time":"2021-05-18T06:01:33Z","timestamp":1621317693000},"page":"616","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["An Outer Membrane Vesicle-Adjuvanted Oral Vaccine Protects Against Lethal, Oral Salmonella Infection"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2194-3978","authenticated-orcid":false,"given":"Jaikin E.","family":"Harrell","sequence":"first","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan R.","family":"Kurtz","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David L.","family":"Bauer","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Timothy","family":"Prior","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick S.","family":"Gellings","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4799-4941","authenticated-orcid":false,"given":"Lisa A.","family":"Morici","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James B.","family":"McLachlan","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Tulane University School of Medicine, New Orleans, LA 70112, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2021,5,18]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"20130433","DOI":"10.1098\/rstb.2013.0433","article-title":"The contribution of vaccination to global health: Past, present and future","volume":"369","author":"Greenwood","year":"2014","journal-title":"Philos. Trans. R. Soc. B Biol. Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1016\/j.immuni.2010.10.002","article-title":"Vaccine Adjuvants: Putting Innate Immunity to Work","volume":"33","author":"Coffman","year":"2010","journal-title":"Immunity"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"148","DOI":"10.3390\/vaccines3010148","article-title":"Choice and Design of Adjuvants for Parenteral and Mucosal Vaccines","volume":"3","author":"Savelkoul","year":"2015","journal-title":"Vaccines"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.2217\/nnm-2017-0015","article-title":"Recent developments in nanocarrier-aided mucosal vaccination","volume":"12","author":"Kammona","year":"2017","journal-title":"Nanomedicine"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1038\/nri1777","article-title":"Mucosal vaccines: The promise and the challenge","volume":"6","author":"Neutra","year":"2006","journal-title":"Nat. Rev. Immunol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1016\/j.addr.2015.03.007","article-title":"Delivery strategies to enhance oral vaccination against enteric infections","volume":"91","author":"Davitt","year":"2015","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"2645","DOI":"10.1101\/gad.1299905","article-title":"Bacterial outer membrane vesicles and the host-pathogen interaction","volume":"19","author":"Kuehn","year":"2005","journal-title":"Genes Dev."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1053","DOI":"10.3389\/fmicb.2017.01053","article-title":"Outer Membrane Vesicles (OMVs) of Gram-negative Bacteria: A Perspective Update","volume":"8","author":"Jan","year":"2017","journal-title":"Front. Microbiol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"B3","DOI":"10.1016\/j.vaccine.2009.04.071","article-title":"Properties and clinical performance of vaccines containing outer membrane vesicles from Neisseria meningitidis","volume":"27","author":"Holst","year":"2009","journal-title":"Vaccine"},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Baker, S.M., Davitt, C.J.H., Motyka, N., Kikendall, N.L., Russell-Lodrigue, K., Roy, C.J., and Morici, L.A. (2017). A Burkholderia pseudomallei Outer Membrane Vesicle Vaccine Provides Cross Protection against Inhalational Glanders in Mice and Non-Human Primates. Vaccines, 5.","DOI":"10.3390\/vaccines5040049"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"8381","DOI":"10.1016\/j.vaccine.2011.08.058","article-title":"A naturally derived outer-membrane vesicle vaccine protects against lethal pulmonary Burkholderia pseudomallei infection","volume":"29","author":"Nieves","year":"2011","journal-title":"Vaccine"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1128\/CVI.00119-14","article-title":"A Burkholderia pseudomallei Outer Membrane Vesicle Vaccine Provides Protection against Lethal Sepsis","volume":"21","author":"Nieves","year":"2014","journal-title":"Clin. Vaccine Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1016\/j.provac.2014.07.007","article-title":"Evaluation of a Burkholderia Pseudomallei Outer Membrane Vesicle Vaccine in Nonhuman Primates","volume":"8","author":"Petersen","year":"2014","journal-title":"Procedia Vaccinol."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Prior, J., Davitt, C., Kurtz, J., Gellings, P., McLachlan, J., and Morici, L. (2021). Bacterial-Derived Outer Membrane Vesicles Are Potent Adjuvants That Drive Humoral and Cellular Immune Responses. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13020131"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"4554","DOI":"10.1128\/IAI.00532-08","article-title":"Immunization with Vibrio cholerae Outer Membrane Vesicles Induces Protective Immunity in Mice","volume":"76","author":"Schild","year":"2008","journal-title":"Infect. Immun."},{"key":"ref_16","first-page":"391","article-title":"Same species, different diseases: How and why typhoidal and non-typhoidal Salmonella enterica serovars differ","volume":"5","author":"Boyle","year":"2014","journal-title":"Front. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2209","DOI":"10.1056\/NEJMoa1905047","article-title":"Phase 3 Efficacy Analysis of a Typhoid Conjugate Vaccine Trial in Nepal","volume":"381","author":"Shakya","year":"2019","journal-title":"N. Engl. J. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1509","DOI":"10.1586\/14760584.2015.1081067","article-title":"Effect of vaccine administration modality on immunogenicity and efficacy","volume":"14","author":"Zhang","year":"2015","journal-title":"Expert Rev. Vaccines"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"986","DOI":"10.4049\/jimmunol.164.2.986","article-title":"Characterization of CD4+T Cell Responses During Natural Infection withSalmonella typhimurium","volume":"164","author":"McSorley","year":"2000","journal-title":"J. Immunol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2424","DOI":"10.1128\/IAI.00052-14","article-title":"Vaccination with a Single CD4 T Cell Peptide Epitope from a Salmonella Type III-Secreted Effector Protein Provides Protection against Lethal Infection","volume":"82","author":"Kurtz","year":"2014","journal-title":"Infect. Immun."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Kurtz, J.R., Nieves, W., Bauer, D.L., Israel, K.E., Adcox, H.E., Gunn, J.S., Morici, L.A., and McLachlan, J.B. (2020). Salmonella Persistence and Host Immunity Are Dictated by the Anatomical Microenvironment. Infect. Immun., 88.","DOI":"10.1128\/IAI.00026-20"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1128\/MMBR.00007-17","article-title":"Vaccination against Salmonella Infection: The Mucosal Way","volume":"81","author":"Gayet","year":"2017","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"624622","DOI":"10.3389\/fcimb.2020.624622","article-title":"Salmonella Biofilm Formation, Chronic Infection, and Immunity Within the Intestine and Hepatobiliary Tract","volume":"10","author":"Harrell","year":"2021","journal-title":"Front. Cell. Infect. Microbiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/nri3802","article-title":"Memory B cells","volume":"15","author":"Kurosaki","year":"2015","journal-title":"Nat. Rev. Immunol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"450","DOI":"10.1111\/j.1365-2567.2005.02140.x","article-title":"Tracking the dynamics of T-cell activation in response to Salmonella infection","volume":"114","author":"Ravindran","year":"2005","journal-title":"Immunology"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"11241","DOI":"10.1073\/pnas.0902015106","article-title":"Positive selection optimizes the number and function of MHCII-restricted CD4+ T cell clones in the naive polyclonal repertoire","volume":"106","author":"Chu","year":"2009","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"623","DOI":"10.4049\/jimmunol.1601686","article-title":"Cutting Edge: Adenosine A2a Receptor Signals Inhibit Germinal Center T Follicular Helper Cell Differentiation during the Primary Response to Vaccination","volume":"198","author":"Schmiel","year":"2016","journal-title":"J. Immunol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"5503","DOI":"10.4049\/jimmunol.1201413","article-title":"Cutting Edge: B Cells Are Essential for Protective Immunity againstSalmonellaIndependent of Antibody Secretion","volume":"189","author":"Nanton","year":"2012","journal-title":"J. Immunol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"230","DOI":"10.1111\/imr.12400","article-title":"Peyer\u2019s patches: Organizing B-cell responses at the intestinal frontier","volume":"271","author":"Reboldi","year":"2016","journal-title":"Immunol. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1016\/j.cell.2010.10.032","article-title":"Germinal Center Dynamics Revealed by Multiphoton Microscopy with a Photoactivatable Fluorescent Reporter","volume":"143","author":"Victora","year":"2010","journal-title":"Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1016\/S1473-3099(19)30418-9","article-title":"The global burden of non-typhoidal salmonella invasive disease: A systematic analysis for the Global Burden of Disease Study 2017","volume":"19","author":"Stanaway","year":"2019","journal-title":"Lancet Infect. Dis."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"901","DOI":"10.1128\/CMR.00002-15","article-title":"Epidemiology, Clinical Presentation, Laboratory Diagnosis, Antimicrobial Resistance, and Antimicrobial Management of Invasive Salmonella Infections","volume":"28","author":"Crump","year":"2015","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"941","DOI":"10.3201\/eid2106.140999","article-title":"Global Burden of Invasive NontyphoidalSalmonellaDisease, 20101","volume":"21","author":"Ao","year":"2015","journal-title":"Emerg. Infect. Dis."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2489","DOI":"10.1016\/S0140-6736(11)61752-2","article-title":"Invasive non-typhoidal salmonella disease: An emerging and neglected tropical disease in Africa","volume":"379","author":"Feasey","year":"2012","journal-title":"Lancet"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1016\/j.jconrel.2016.02.014","article-title":"Mucosal vaccine delivery: Current state and a pediatric perspective","volume":"240","author":"Shakya","year":"2016","journal-title":"J. Control. Release"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"116","DOI":"10.1016\/j.addr.2017.04.008","article-title":"Current state and challenges in developing oral vaccines","volume":"114","author":"Ramirez","year":"2017","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1038\/s41541-021-00281-z","article-title":"Burkholderia pseudomallei OMVs derived from infection mimicking conditions elicit similar protection to a live-attenuated vaccine","volume":"6","author":"Baker","year":"2021","journal-title":"NPJ Vaccines"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"S22","DOI":"10.1016\/S0264-410X(99)00231-5","article-title":"Duration of efficacy of Ty21a, attenuated Salmonella typhi live oral vaccine","volume":"17","author":"Levine","year":"1999","journal-title":"Vaccine"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"e01421-15","DOI":"10.1128\/mBio.01421-15","article-title":"A Highly Effective Component Vaccine against Nontyphoidal Salmonella enterica Infections","volume":"6","author":"Ferreira","year":"2015","journal-title":"MBio"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"4175","DOI":"10.1128\/IAI.05278-11","article-title":"Engineering and Preclinical Evaluation of Attenuated Nontyphoidal Salmonella Strains Serving as Live Oral Vaccines and as Reagent Strains","volume":"79","author":"Tennant","year":"2011","journal-title":"Infect. Immun."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1016\/j.ijmm.2016.08.004","article-title":"Outer membrane vesicles derived from Salmonella Typhimurium mutants with truncated LPS induce cross-protective immune responses against infection of Salmonella enterica serovars in the mouse model","volume":"306","author":"Liu","year":"2016","journal-title":"Int. J. Med. Microbiol."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1128\/mr.47.1.46-83.1983","article-title":"Bactericidal and Bacteriolytic Activity of Serum against Gram-Negative Bacteria","volume":"47","author":"Taylor","year":"1983","journal-title":"Microbiol. Rev."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"102","DOI":"10.1016\/S0966-842X(98)01208-6","article-title":"Antibody-mediated protection against intracellular pathogens","volume":"6","author":"Casadevall","year":"1998","journal-title":"Trends Microbiol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"5879","DOI":"10.1016\/j.vaccine.2013.09.041","article-title":"Safety and tolerability of a live oral Salmonella typhimurium vaccine candidate in SIV-infected nonhuman primates","volume":"31","author":"Ault","year":"2013","journal-title":"Vaccine"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1553","DOI":"10.1172\/JCI33998","article-title":"The neglected role of antibody in protection against bacteremia caused by nontyphoidal strains of Salmonella in African children","volume":"118","author":"MacLennan","year":"2008","journal-title":"J. Clin. Investig."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1016\/0016-5085(94)90214-3","article-title":"Monoclonal immunoglobulin a prevents adherence and invasion of polarized epithelial cell monolayers by Salmonella typhimurium","volume":"107","author":"Michetti","year":"1994","journal-title":"Gastroenterology"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1016\/0882-4010(87)90019-2","article-title":"Role of monoclonal O-antigen antibody epitope specificity and isotype in protection against experimental mouse typhoid","volume":"2","author":"Carlin","year":"1987","journal-title":"Microb. Pathog."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"S537","DOI":"10.1093\/clinids\/9.Supplement_5.S537","article-title":"The Function of Antibody and Complement in the Lysis of Bacteria","volume":"9","author":"Frank","year":"1987","journal-title":"Clin. Infect. Dis."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"436","DOI":"10.1093\/infdis\/150.3.436","article-title":"Reexamination of the Protective Role of the Capsular Polysaccharide (Vi antigen) of Salmonella typhi","volume":"150","author":"Robbins","year":"1984","journal-title":"J. Infect. Dis."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"120","DOI":"10.1016\/j.immuni.2015.06.013","article-title":"Salmonella Infection Drives Promiscuous B Cell Activation Followed by Extrafollicular Affinity Maturation","volume":"43","author":"Lee","year":"2015","journal-title":"Immunity"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"3136","DOI":"10.1128\/IAI.01313-09","article-title":"A Burkholderia pseudomallei \u0394purM Mutant Is Avirulent in Immunocompetent and Immunodeficient Animals: Candidate Strain for Exclusion from Select-Agent Lists","volume":"78","author":"Propst","year":"2010","journal-title":"Infect. Immun."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"565","DOI":"10.1038\/nprot.2009.9","article-title":"Tracking epitope-specific T cells","volume":"4","author":"Moon","year":"2009","journal-title":"Nat. Protoc."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"2065","DOI":"10.1084\/jem.20112272","article-title":"Deletion and anergy of polyclonal B cells specific for ubiquitous membrane-bound self-antigen","volume":"209","author":"Taylor","year":"2012","journal-title":"J. Exp. Med."}],"container-title":["Pathogens"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/5\/616\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T06:03:02Z","timestamp":1760162582000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2076-0817\/10\/5\/616"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,5,18]]},"references-count":53,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["pathogens10050616"],"URL":"https:\/\/doi.org\/10.3390\/pathogens10050616","relation":{},"ISSN":["2076-0817"],"issn-type":[{"value":"2076-0817","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,18]]}}}