In vitro modeling of polyclonal infection dynamics within the human airways by Haemophilus influenzae differential fluorescent labeling

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dc.contributor.authorRapún Araiz, Beatriz
dc.contributor.authorSorzabal-Bellido, Ioritz
dc.contributor.authorAsensio López, Javier
dc.contributor.authorLázaro-Díez, María
dc.contributor.authorAriz Galilea, Mikel
dc.contributor.authorSobejano de la Merced, Carlos
dc.contributor.authorEuba, Begoña
dc.contributor.authorFernández Calvet, Ariadna
dc.contributor.authorCortés Domínguez, Iván
dc.contributor.authorBurgui Erice, Saioa
dc.contributor.authorToledo Arana, Alejandro
dc.contributor.authorOrtiz de Solórzano, Carlos
dc.contributor.authorGarmendia García, Juncal
dc.contributor.departmentIngeniería Eléctrica, Electrónica y de Comunicaciónes_ES
dc.contributor.departmentIngeniaritza Elektrikoa, Elektronikoa eta Telekomunikazio Ingeniaritzaeu
dc.contributor.departmentInstitute of Smart Cities - ISCen
dc.date.accessioned2024-05-27T09:34:54Z
dc.date.available2024-05-27T09:34:54Z
dc.date.issued2023
dc.date.updated2024-05-27T09:16:58Z
dc.description.abstractStandardized clinical procedures for antibiotic administration rely on pathogen identification and antibiotic susceptibility testing, often performed on single-colony bacterial isolates. For respiratory pathogens, this could be questionable, as chronic patients may be persistently colonized by multiple clones or lineages from the same bacterial pathogen species. Indeed, multiple strains of nontypeable Haemophilus influenzae, with different antibiotic susceptibility profiles, can be co-isolated from cystic fibrosis and chronic obstructive pulmonary disease sputum specimens. Despite this clinical evidence, we lack information about the dynamics of H. influenzae polyclonal infections, which limits the optimization of therapeutics. Here, we present the engineering and validation of a plasmid toolkit (pTBH, toolbox for Haemophilus), with standardized modules consisting of six reporter genes for fluorescent or bioluminescent labeling of H. influenzae. This plasmid set was independently introduced in a panel of genomically and phenotypically different H. influenzae strains, and two of them were used as a proof of principle to analyze mixed biofilm growth architecture and antibiotic efficacy, and to visualize the dynamics of alveolar epithelial co-infection. The mixed biofilms showed a bilayer architecture, and antibiotic efficacy correlated with the antibiotic susceptibility of the respective single-species strains. Furthermore, differential kinetics of bacterial intracellular location within subcellular acidic compartments were quantified upon co-infection of cultured airway epithelial cells. Overall, we present a panel of novel plasmid tools and quantitative image analysis methods with the potential to be used in a whole range of bacterial host species, assay types, and¿or conditions and generate meaningful information for clinically relevant settings.en
dc.description.sponsorshipJ.A.-L. is funded by a PhD studentship from Regional Navarra Government, Spain, reference 0011-1408-2020-000007. C.S.M. is funded by a Formación de Profesorado Universitario PhD studentship from the Spanish Ministry of Science and Innovation (MCINN), Spain, reference FPU20/06252. This work has been funded by grants from Ministerio de Ciencia, Innovación y Universidades, Agencia Estatal de Investigación (MCIU/AEI/10.13039/50110011033) and FEDER funds EU, RTI2018-094494-BC22, PDI2021-122409OB-C22 (C.O.S.), and RTI2018-096369-B-I00, PID2021-125947OB-I00 (J.G.); from SEPAR, 875/2019 (J.G.); from Gobierno de Navarra, PC150 and PC136 (J.G.) and PC151 and PC137 (C.O.S.). CIBER is an initiative from Instituto de Salud Carlos III, Madrid, Spain. Experimental design: B.R.A., I.S.B., M.L.D., A.T.A., C.O.S., J.G.; experimental work: B.R.A., I.S.B., J.A.L., B.E., M.L.D., C.O.M., A.F.C.; data analysis: B.R.A., I.S.B., M.L.D., M.A.; writing of the manuscript: J.G., C.O.S.; correction of the manuscript: all authors; funding¿ I.C.D., S.B., C.O.S., J.G.; Funding text 2: J.A.-L. is funded by a PhD studentship from Regional Navarra Govern, Spain, reference 0011-1408-2020-000007. C.S.M. is funded by a Formación de Profesorado Universitario PhD studentship from the Spanish Ministry of Science and Innovation (MCINN), Spain, reference FPU20/06252. This work has been funded by grants from Ministerio de Ciencia, Innovación y Universidades, Agencia Estatal de Investigación (MCIU/AEI/10.13039/50110011033) and FEDER funds EU, RTI2018-094494-BC22, PDI2021-122409OB-C22 (C.O.S.), and RTI2018-096369-B-I00, PID2021-125947OB-I00 (J.G.); from SEPAR, 875/2019 (J.G.); from Gobierno de Navarra, PC150 and PC136 (J.G.) and PC151 and PC137 (C.O.S.). CIBER is an initiative from Instituto de Salud Carlos III, Madrid, Spain.en
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dc.identifier.citationRapún-Araiz, B., Sorzabal-Bellido, I., Asensio-López, J., Lázaro-Díez, M., Ariz, M., Sobejano de la Merced, C., Euba, B., Fernández-Calvet, A., Cortés-Domínguez, I., Burgui, S., Toledo-Arana, A., Ortiz-de-Solórzano, C., Garmendia, J. (2023) In vitro modeling of polyclonal infection dynamics within the human airways by Haemophilus influenzae differential fluorescent labeling. Microbiology Spectrum, 11(6), 1-20. https://doi.org/10.1128/spectrum.00993-23.es_ES
dc.identifier.doi10.1128/spectrum.00993-23
dc.identifier.issn2165-0497
dc.identifier.urihttps://academica-e.unavarra.es/handle/2454/48193
dc.language.isoengen
dc.publisherAmerican Society for Microbiologyen
dc.relation.ispartofMicrobiology Spectrum (2023), vol. 11(6)es_ES
dc.relation.projectIDinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-094494-B-C22/ES/
dc.relation.projectIDinfo:eu-repo/grantAgreement///PDI2021-122409OB-C22/
dc.relation.projectIDinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-096369-B-I00/ES/
dc.relation.projectIDinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2021-2023/PID2021-125947OB-I00/ES/
dc.relation.projectIDinfo:eu-repo/grantAgreement/Gobierno de Navarra//PC150/
dc.relation.projectIDinfo:eu-repo/grantAgreement/Gobierno de Navarra//PC136/
dc.relation.projectIDinfo:eu-repo/grantAgreement/Gobierno de Navarra//PC151/
dc.relation.projectIDinfo:eu-repo/grantAgreement/Gobierno de Navarra//PC137/
dc.relation.publisherversionhttps://doi.org/10.1128/spectrum.00993-23
dc.rights© 2023 Rapún-Araiz et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.en
dc.rights.accessRightsinfo:eu-repo/semantics/openAccess
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectFluorescent labelingen
dc.subjectHaemophilus influenzaeen
dc.subjectMixed biofilmsen
dc.subjectMixed epithelial infectionen
dc.subjectPlasmid toolboxen
dc.subjectPolyclonality assessmenten
dc.subjectSelective antibiotic efficacyen
dc.titleIn vitro modeling of polyclonal infection dynamics within the human airways by Haemophilus influenzae differential fluorescent labelingen
dc.typeinfo:eu-repo/semantics/article
dc.type.versioninfo:eu-repo/semantics/acceptedVersion
dspace.entity.typePublication
relation.isAuthorOfPublication45523631-0bc0-4010-b999-22ab4f7ef380
relation.isAuthorOfPublication9d2b9c4c-9ede-4367-9ad9-8987addfbff6
relation.isAuthorOfPublicationeab5adde-437b-46f6-9496-2fa2b16b308f
relation.isAuthorOfPublication37fcbeeb-a960-4b98-9677-3d84608f2be1
relation.isAuthorOfPublication777af947-83d1-4cb7-b485-b0430d3e28be
relation.isAuthorOfPublication.latestForDiscovery45523631-0bc0-4010-b999-22ab4f7ef380

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