dc.contributor.author
Butkevych, Eduard
dc.contributor.author
Lobo de Sá, Fábia Daniela
dc.contributor.author
Nattramilarasu, Praveen Kumar
dc.contributor.author
Bücker, Roland
dc.date.accessioned
2020-04-20T07:44:07Z
dc.date.available
2020-04-20T07:44:07Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/27123
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-26883
dc.description.abstract
Campylobacter jejuni is a widespread zoonotic pathogen and the leading bacterial cause of foodborne gastroenteritis in humans. Previous infection studies showed disruption of intercellular contacts, induction of epithelial apoptosis, and immune activation, all three contributing to intestinal barrier dysfunction leading to diarrhea. The present study aims to determine the impact of subepithelial immune cells on intestinal barrier dysfunction during Campylobacter jejuni infection and the underlying pathological mechanisms. Infection was performed in a co-culture of confluent monolayers of the human colon cell line HT-29/B6-GR/MR and THP-1 immune cells. Twenty-two hours after infection, transepithelial electrical resistance (TER) was decreased by 58 ± 6% compared to controls. The infection resulted in an increase in permeability for fluorescein (332 Da; 4.5-fold) and for FITC-dextran (4 kDa; 3.5-fold), respectively. In contrast, incubation of the co-culture with the pan-caspase inhibitor Q-VD-OPh during the infection resulted in a complete recovery of the decrease in TER and a normalization of flux values. Fluorescence microscopy showed apoptotic fragmentation in infected cell monolayers resulting in a 5-fold increase of the apoptotic ratio, accompanied by an increased caspase-3 cleavage and caspase-3/7 activity, which both were not present after Q-VD-OPh treatment. Western blot analysis revealed increased claudin-1 and claudin-2 protein expression. Inhibition of apoptosis induction did not normalize these tight junction changes. TNFα concentration was increased during the infection in the co-culture. In conclusion, Campylobacter jejuni infection and the consequent subepithelial immune activation cause intestinal barrier dysfunction mainly through caspase-3-dependent epithelial apoptosis. Concomitant tight junction changes were caspase-independent. Anti-apoptotic and immune-modulatory substances appear to be promising agents for treatment of campylobacteriosis.
en
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
epithelial barrier
en
dc.subject
tumor necrosis factor alpha
en
dc.subject
Campylobacter
en
dc.subject
epithelial cell
en
dc.subject
immune cell co-culture
en
dc.subject
tight junction
en
dc.subject.ddc
600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::610 Medizin und Gesundheit
dc.title
Contribution of Epithelial Apoptosis and Subepithelial Immune Responses in Campylobacter jejuni-Induced Barrier Disruption
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.articlenumber
344
dcterms.bibliographicCitation.doi
10.3389/fmicb.2020.00344
dcterms.bibliographicCitation.journaltitle
Frontiers in Microbiology
dcterms.bibliographicCitation.originalpublishername
Frontiers Media S.A.
dcterms.bibliographicCitation.volume
11
refubium.affiliation
Charité - Universitätsmedizin Berlin
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.bibliographicCitation.pmid
32210941
dcterms.isPartOf.eissn
1664-302X