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dc.contributor.authorHaag, Andreas
dc.contributor.authorLiljeroos, Lassi
dc.contributor.authorDonato, Paolo
dc.contributor.authorPozzi, Clarissa
dc.contributor.authorBrignoli, Tarcisio
dc.contributor.authorBottomley, Matthew J.
dc.contributor.authorBagnoli, Fabio
dc.contributor.authorDelany, Isabel
dc.date.accessioned2023-01-24T16:30:09Z
dc.date.available2023-01-24T16:30:09Z
dc.date.issued2023-01-23
dc.identifier.citationHaag , A , Liljeroos , L , Donato , P , Pozzi , C , Brignoli , T , Bottomley , M J , Bagnoli , F & Delany , I 2023 , ' In vivo  gene expression profiling of Staphylococcus aureus during infection informs design of stemless leukocidins LukE and -D as detoxified vaccine candidates ' , Microbiology Spectrum , vol. Ahead of Print , e02574-22 . https://doi.org/10.1128/spectrum.02574-22en
dc.identifier.issn2165-0497
dc.identifier.otherPURE: 283069759
dc.identifier.otherPURE UUID: 7958a227-30ce-474f-919b-bfc6f938b082
dc.identifier.otherRIS: urn:D1075C5A9F230F7DAAE48FABB1C9E4CE
dc.identifier.urihttp://hdl.handle.net/10023/26822
dc.descriptionA.F.H. and L.L. were funded by Marie Sklodowska-Curie Intra-European Fellowships (PIEF-GA-2012-328377 and PIEF-GA-2013-623168, respectively).en
dc.description.abstractStaphylococcus aureus is a clinically important bacterial pathogen that has become resistant to treatment with most routinely used antibiotics. Alternative strategies, such as vaccination and phage therapy, are therefore actively being investigated to prevent or combat staphylococcal infections. Vaccination requires that vaccine targets are expressed at sufficient quantities during infection so that they can be targeted by the host’s immune system. While our knowledge of in vitro expression levels of putative vaccine candidates is comprehensive, crucial in vivo expression data are scarce and promising vaccine candidates during in vitro assessment often prove ineffective in preventing S. aureus infection. Here, we show how a newly developed high-throughput quantitative reverse transcription-PCR (qRT-PCR) assay monitoring the expression of 84 staphylococcal genes encoding mostly virulence factors can inform the selection and design of effective vaccine candidates against staphylococcal infections. We show that this assay can accurately quantify mRNA expression levels of these genes in several host organs relying only on very limited amounts of bacterial mRNA in each sample. We selected two highly expressed genes, lukE and lukD, encoding pore-forming leukotoxins, to inform the design of detoxified recombinant proteins and showed that immunization with recombinant genetically detoxified LukED antigens conferred protection against staphylococcal skin infection in mice. Consequently, knowledge of in vivo-expressed virulence determinants can be successfully deployed to identify and select promising candidates for optimized design of effective vaccine antigens against S. aureus. Notably, this approach should be broadly applicable to numerous other pathogens.
dc.format.extent19
dc.language.isoeng
dc.relation.ispartofMicrobiology Spectrumen
dc.rightsCopyright © 2023 Haag et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.en
dc.subjectIn vivo gene expressionen
dc.subjectHigh-throughput qRT-PCRen
dc.subjectStaphylococcus aureusen
dc.subjectStaphylococcus aureus vaccineen
dc.subjectLeukocidinsen
dc.subjectQR Microbiologyen
dc.subjectRA0421 Public health. Hygiene. Preventive Medicineen
dc.subjectDASen
dc.subject.lccQRen
dc.subject.lccRA0421en
dc.titleIn vivo gene expression profiling of Staphylococcus aureus during infection informs design of stemless leukocidins LukE and -D as detoxified vaccine candidatesen
dc.typeJournal articleen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. School of Medicineen
dc.identifier.doihttps://doi.org/10.1128/spectrum.02574-22
dc.description.statusPeer revieweden


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