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Krsek, Daniel, Yara, Daniel Alejandro, Hrbáčková, Hana, Daniel, Ondřej, Mančíková, Andrea, Schüller, Stephanie 
ORCID: https://orcid.org/0000-0003-3260-9112 and Bielaszewska, Martina
(2023)
Translocation of outer membrane vesicles from enterohemorrhagic Escherichia coli O157 across the intestinal epithelial barrier.
Frontiers in Microbiology, 14.
ISSN 1664-302X
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Abstract
Outer membrane vesicles (OMVs) carrying virulence factors of enterohemorrhagic Escherichia coli (EHEC) are assumed to play a role in the pathogenesis of life-threatening hemolytic uremic syndrome (HUS). However, it is unknown if and how OMVs, which are produced in the intestinal lumen, cross the intestinal epithelial barrier (IEB) to reach the renal glomerular endothelium, the major target in HUS. We investigated the ability of EHEC O157 OMVs to translocate across the IEB using a model of polarized Caco-2 cells grown on Transwell inserts and characterized important aspects of this process. Using unlabeled or fluorescently labeled OMVs, tests of the intestinal barrier integrity, inhibitors of endocytosis, cell viability assay, and microscopic techniques, we demonstrated that EHEC O157 OMVs translocated across the IEB. OMV translocation involved both paracellular and transcellular pathways and was significantly increased under simulated inflammatory conditions. In addition, translocation was not dependent on OMV-associated virulence factors and did not affect viability of intestinal epithelial cells. Importantly, translocation of EHEC O157 OMVs was confirmed in human colonoids thereby supporting physiological relevance of OMVs in the pathogenesis of HUS.
| Item Type: | Article |
|---|---|
| Additional Information: | Funding information: This study was supported by the Grant Agency of the Czech Republic (GACR), grant number: 21-06792S (to MB) and the UKRI Biotechnology and Biological Sciences Research Council (BBSRC) (Norwich Research Park Biosciences Doctoral Training studentship BB/M011216/1 to DY). Data availability statement: The original contributions presented in this study are included in the article/Supplementary material, further inquiries can be directed to the corresponding author. |
| Faculty \ School: | Faculty of Medicine and Health Sciences > Norwich Medical School |
| UEA Research Groups: | Faculty of Medicine and Health Sciences > Research Groups > Gastroenterology and Gut Biology Faculty of Medicine and Health Sciences > Research Centres > Metabolic Health Faculty of Medicine and Health Sciences > Research Groups > Pathogen Biology Group |
| Depositing User: | LivePure Connector |
| Date Deposited: | 07 Jun 2023 00:05 |
| Last Modified: | 12 Nov 2024 00:49 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/92300 |
| DOI: | 10.3389/fmicb.2023.1198945 |
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