Structural Insights into Escherichia coli Shiga Toxin (Stx) Glycosphingolipid Receptors of Porcine Renal Epithelial Cells and Inhibition of Stx-Mediated Cellular Injury Using Neoglycolipid-Spiked Glycovesicles

Shiga toxin (Stx) producing Escherichia coli (STEC) cause the edema disease in pigs by releasing the swine-pathogenic Stx2e subtype as the key virulence factor. Stx2e targets endothelial cells of animal organs including the kidney harboring the Stx receptor glycosphingolipids (GSLs) globotriaosylcer...

Authors: Detzner, Johanna
Gloerfeld, Caroline
Pohlentz, Gottfried
Legros, Nadine
Humpf, Hans-Ulrich
Mellmann, Alexander
Karch, Helge
Müthing, Johannes
Division/Institute:FB 05: Medizinische Fakultät
Document types:Article
Media types:Text
Publication date:2019
Date of publication on miami:06.12.2019
Modification date:27.01.2020
Edition statement:[Electronic ed.]
Source:Microorganisms 7 (2019) 11, 582, 1-25
Subjects:edema disease; epithelial cells; Gb3Cer; Gb4Cer; glycosphingolipids; LLC-PK1; neoglycolipids; PK-15; porcine kidney; Stx2e
DDC Subject:610: Medizin und Gesundheit
License:CC BY 4.0
Language:English
Funding:Finanziert durch den Open-Access-Publikationsfonds der Westfälischen Wilhelms-Universität Münster (WWU Münster).
This research was supported by grants from the German Research Foundation (DFG), grant number MU845/7-1 with reference number 404813761 (J.M.) and project B04 of the SFB 1009 (A.M. and H.K.), and the German Federal Ministry of Education and Research (BMBF), conducted under the umbrella of the German Center for Infection Research (DZIF, TTU 06.801) with assistance of InfectControl 2020 (TFP-TV8-AS12, ref. no. 03ZZ0802H) and InfectControl 2020 IRMRESS (ref. no. 03ZZ0805B).
Förderer: Deutsche Forschungsgemeinschaft / Projektnummer: 404813761
Format:PDF document
URN:urn:nbn:de:hbz:6-42199522061
Permalink:http://nbn-resolving.de/urn:nbn:de:hbz:6-42199522061
Other Identifiers:DOI: 10.3390/microorganisms7110582
Digital documents:artikel_muething_2019.pdf
zusatzmaterial_muething_2019.pdf
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Shiga toxin (Stx) producing Escherichia coli (STEC) cause the edema disease in pigs by releasing the swine-pathogenic Stx2e subtype as the key virulence factor. Stx2e targets endothelial cells of animal organs including the kidney harboring the Stx receptor glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer, Galα1-4Galβ1-4Glcβ1-1Cer) and globotetraosylceramide (Gb4Cer, GalNAcβ1-3Galα1-4Galβ1-4Glcβ1-1Cer). Since the involvement of renal epithelial cells in the edema disease is unknown, in this study, we analyzed the porcine kidney epithelial cell lines, LLC-PK1 and PK-15, regarding the presence of Stx-binding GSLs, their sensitivity towards Stx2e, and the inhibitory potential of Gb3- and Gb4-neoglycolipids, carrying phosphatidylethanolamine (PE) as the lipid anchor, towards Stx2e. Immunochemical and mass spectrometric analysis revealed various Gb3Cer and Gb4Cer lipoforms as the dominant Stx-binding GSLs in both LLC-PK1 and PK-15 cells. A dihexosylceramide with proposed Galα1-4Gal-sequence (Gal2Cer) was detected in PK-15 cells, whereas LLC-PK1 cells lacked this compound. Both cell lines were susceptible towards Stx2e with LLC-PK1 representing an extremely Stx2e-sensitive cell line. Gb3-PE and Gb4-PE applied as glycovesicles significantly reduced the cytotoxic activity of Stx2e towards LLC-PK1 cells, whereas only Gb4-PE exhibited some protection against Stx2e for PK-15 cells. This is the first report identifying Stx2e receptors of porcine kidney epithelial cells and providing first data on their Stx2e-mediated damage suggesting possible involvement in the edema disease.