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Outer Membrane Vesicles From Brucella melitensis Modulate Immune Response and Induce Cytoskeleton Rearrangement in Peripheral Blood Mononuclear Cells

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dc.contributor.authorAvila-Calderon, Eric Danielen
dc.contributor.authorMedina-Chavez, Olinen
dc.contributor.authorFlores-Romo, Leopoldoen
dc.contributor.authorHernandez-Hernandez, Jose Manuelen
dc.contributor.authorDonis-Maturano, Luisen
dc.contributor.authorLopez-Merino, Ahidéen
dc.contributor.authorArellano-Reynoso, Beatrizen
dc.contributor.authorAguilera-Arreola, Ma. Guadalupeen
dc.contributor.authorRuiz, Enrico A.en
dc.contributor.authorGomez-Lunar, Zulemaen
dc.contributor.authorWitonsky, Sharon G.en
dc.contributor.authorContreras-Rodriguez, Aracelien
dc.contributor.departmentLarge Animal Clinical Sciencesen
dc.date.accessioned2021-01-29T15:24:05Zen
dc.date.available2021-01-29T15:24:05Zen
dc.date.issued2020-10-19en
dc.description.abstractSimilar to what has been described in other Gram-negative bacteria, Brucella melitensis releases outer membrane vesicles (OMVs). OMVs from B. melitensis 16M and the rough-mutant B. melitensis VTRM1 were able to induce a protective immune response against virulent B. melitensis in mice models. The presence of some proteins which had previously been reported to induce protection against Brucella were found in the proteome of OMVs from B. melitensis 16M. However, the proteome of OMVs from B. melitensis VTRM1 had not previously been determined. In order to be better understand the role of OMVs in host-cell interactions, the aim of this work was to compare the proteomes of OMVs from B. melitensis 16M and the derived rough-mutant B. melitensis VTRM1, as well as to characterize the immune response induced by vesicles on host cells. Additionally, the effect of SDS and proteinase K on the stability of OMVs was analyzed. OMVs from B. melitensis 16M (smooth strain) and the B. melitensis VTRM1 rough mutant (lacking the O-polysaccharide side chain) were analyzed through liquid chromatography-mass spectrometry (LC-MS/MS). OMVs were treated with proteinase K, sodium deoxycholate, and SDS, and then their protein profile was determined using SDS-PAGE. Furthermore, PBMCs were treated with OMVs in order to measure their effect on cytoskeleton, surface molecules, apoptosis, DNA damage, proliferation, and cytokine-induction. A total of 131 proteins were identified in OMVs from B. melitensis16M, and 43 in OMVs from B. melitensis VTRM1. Proteome comparison showed that 22 orthologous proteins were common in vesicles from both strains, and their core proteome contained Omp31, Omp25, GroL, and Omp16. After a subsequent detergent and enzyme treatment, OMVs from B. melitensis VTRM1 exhibited higher sensitive compared to OMVs from the B. melitensis 16M strain. Neither OMVs induced IL-17, proliferation, apoptosis or DNA damage. Nonetheless, OMVs from the smooth and rough strains induced overproduction of TNF alpha and IL-6, as well as actin and tubulin rearrangements in the cytoskeleton. Moreover, OMVs from both strains inhibited PD-L1 expression in T-cells. These data revealed significant differences in OMVs derived from the rough and smooth Brucella strains, among which, the presence or absence of complete LPS appeared to be crucial to protect proteins contained within vesicles and to drive the immune response.en
dc.description.notesThis work was funded by CONACYT 61529, SIP-IPN 20182152, 20195737, 20200594, SAGARPA-CONACYT 2017-02-291311. EA-C was supported by CONACYT and PIFI-IPN scholarships. MA-A, ER, ZG-L, and AC-R were supported by fellowships from COFAA-IPN, SIP-EDI, and SNI-CONACYT.en
dc.description.sponsorshipCONACYTConsejo Nacional de Ciencia y Tecnologia (CONACyT) [61529]; SIP-IPN [20182152, 20195737, 20200594]; SAGARPA-CONACYTConsejo Nacional de Ciencia y Tecnologia (CONACyT) [2017-02-291311]; CONACYT scholarshipConsejo Nacional de Ciencia y Tecnologia (CONACyT); PIFI-IPN scholarship; COFAA-IPN; SIP-EDI; SNI-CONACYTConsejo Nacional de Ciencia y Tecnologia (CONACyT)en
dc.format.mimetypeapplication/pdfen
dc.identifier.doihttps://doi.org/10.3389/fmicb.2020.556795en
dc.identifier.issn1664-302Xen
dc.identifier.other556795en
dc.identifier.pmid33193138en
dc.identifier.urihttp://hdl.handle.net/10919/102127en
dc.identifier.volume11en
dc.language.isoenen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectouter membrane vesiclesen
dc.subjectBrucellaen
dc.subjectproteomicsen
dc.subjectOMVsen
dc.subjectbacterial vesiclesen
dc.subjectextracellular vesicleen
dc.titleOuter Membrane Vesicles From Brucella melitensis Modulate Immune Response and Induce Cytoskeleton Rearrangement in Peripheral Blood Mononuclear Cellsen
dc.title.serialFrontiers in Microbiologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
dc.type.dcmitypeStillImageen

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