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dc.contributor.authorHerzog, Susann
dc.contributor.authorDach, Felix
dc.contributor.authorde Buhr, Nicole
dc.contributor.authorNiemann, Silke
dc.contributor.authorSchlagowski, Jannik
dc.contributor.authorChaves-Moreno, Diego
dc.contributor.authorNeumann, Claudia
dc.contributor.authorGoretzko, Jonas
dc.contributor.authorSchwierzeck, Vera
dc.contributor.authorMellmann, Alexander
dc.contributor.authorDübbers, Angelika
dc.contributor.authorKüster, Peter
dc.contributor.authorSchültingkemper, Holger
dc.contributor.authorRescher, Ursula
dc.contributor.authorPieper, Dietmar H
dc.contributor.authorVon Köckritz-Blickwede, Maren
dc.contributor.authorKahl, Barbara C
dc.date.accessioned2019-12-10T11:48:05Z
dc.date.available2019-12-10T11:48:05Z
dc.date.issued2019-01-01
dc.identifier.issn1664-3224
dc.identifier.pmid31772562
dc.identifier.doi10.3389/fimmu.2019.02552
dc.identifier.urihttp://hdl.handle.net/10033/622045
dc.description.abstractStaphylococcus aureus is one of the first and most prevalent pathogens cultured from the airways of cystic fibrosis (CF) patients, which can persist there for extended periods. Airway infections in CF patients are characterized by a strong inflammatory response of highly recruited neutrophils. One killing mechanism of neutrophils is the formation of neutrophil extracellular traps (NETs), which capture and eradicate bacteria by extracellular fibers of neutrophil chromatin decorated with antimicrobial granule proteins. S. aureus secretes nuclease, which can degrade NETs. We hypothesized, that S. aureus adapts to the airways of CF patients during persistent infection by escaping from NET-mediated killing via an increase of nuclease activity. Sputum samples of CF patients with chronic S. aureus infection were visualized by confocal microscopy after immuno-fluorescence staining for NET-specific markers, S. aureus bacteria and overall DNA structures. Nuclease activity was analyzed in sequential isogenic long persisting S. aureus isolates, as confirmed by whole genome sequencing, from an individual CF patient using a FRET-based nuclease activity assay. Additionally, some of these isolates were selected and analyzed by qRT-PCR to determine the expression of nuc1 and regulators of interest. NET-killing assays were performed with clinical S. aureus isolates to evaluate killing and bacterial survival depending on nuclease activity. To confirm the role of nuclease during NET-mediated killing, a clinical isolate with low nuclease activity was transformed with a nuclease expression vector (pCM28nuc). Furthermore, two sputa from an individual CF patient were subjected to RNA-sequence analysis to evaluate the activity of nuclease in vivo. In sputa, S. aureus was associated to extracellular DNA structures. Nuclease activity in clinical S. aureus isolates increased in a time-and phenotype-dependent manner. In the clinical isolates, the expression of nuc1 was inversely correlated to the activity of agr and was independent of saeS. NET-mediated killing was significantly higher in S. aureus isolates with low compared to isolates with high nuclease activity. Importantly, transformation of the clinical isolate with low nuclease activity with pCM28nuc conferred protection against NET-mediated killing confirming the beneficial role of nuclease for protection against NETs. Also, nuclease expression in in vivo sputa was high, which underlines the important role of nuclease within the highly inflamed CF airways. In conclusion, our data show that S. aureus adapts to the neutrophil-rich environment of CF airways with increasing nuclease expression most likely to avoid NET-killing during long-term persistence.en_US
dc.language.isoenen_US
dc.publisherFrontiersen_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectCystic fibrosisen_US
dc.subjectNET-mediated killingen_US
dc.subjectStaphylococcus aureusen_US
dc.subjectadaptationen_US
dc.subjectneutrophil extracellular trapsen_US
dc.subjectnucleaseen_US
dc.titleHigh Nuclease Activity of Long Persisting Isolates Within the Airways of Cystic Fibrosis Patients Protects Against NET-Mediated Killing.en_US
dc.typeArticleen_US
dc.identifier.journalFrontiers of Immunologyen_US
refterms.dateFOA2019-12-10T11:48:05Z
dc.source.journaltitleFrontiers in immunology


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