Interaction of Shiga toxin with primary human intestinal and renal epithelial cells: glycovesicles as novel toxin inhibitors
Final Report Abstract
Enterohemorrhagic Escherichia coli (EHEC) are the human pathogenic subset of Shiga toxin (Stx)-producing E. coli (STEC). EHEC are responsible for severe colon infections associated with life-threatening extraintestinal complications such as the hemolytic-uremic syndrome (HUS) and neurological disturbances. Endothelial cells in various human organs are renowned targets of Stx, whereas the role of epithelial cells of colon and kidneys in the infection process are currently discussed intensively. The Stx-binding glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer) and their various lipoforms have been scrutinized in this project for primary human kidney and colon epithelial cells. The mass spectra obtained by electrospray ionization mass spectrometry of Gb3Cer, Gb4Cer, and Gb5Cer of kidney epithelial cells revealed lipoforms carrying sphingosine (d18:1) in the ceramide lipid anchor linked with C16:0, C22:0, or C24:1/C24:0 fatty acyl chains.The dominant Gb3Cer and Gb4Cer lipoforms of colon epithelial cells harbor sphingosine (d18:1) linked to C16:0, C22:1/C22:0, or C24:1/C24:2 fatty acyl chains, whereas Gb3Cer and Gb4Cer lipoforms with C24:0 fatty acyl chains were missing. Collectively, renal epithelial cells exhibit a more complex profile with a higher extent of lipoform variability regarding the Stx receptors Gb3Cer and Gb4Cer and exhibit as a unique feature Gb5Cer, which is undetectable in colonic epithelial cells. The preferential distribution of Gb3Cer and Gb4Cer in lipid raft-analog membrane preparations indicated the association of the Stx receptor GSLs with membrane microdomains. The cytotoxic activity of swine-pathogenic Stx2e subtype towards porcine renal epithelial cells could be reduced using glycovesicles spiked with Gb3- and Gb4-neoglycolipids (neoGLs) carrying a phosphatidylethanolamine (PE) lipid anchor, and the Stx1a- and Stx2a-mediated cytotoxic action against Vero cells could be mitigated using pectin-derived neoGLs of (α1-4)Galn-PE type, which have been produced by us. Our results gave evidence for a large resilience of primary human colon epithelial cells against Stx1a and Stx2a most probably due to their low content of the high-affinity Stx-receptor Gb3Cer. On the other hand, kidney epithelial cells were highly sensitive towards Stx1a and Stx2a. This data suggest that kidney epithelial cells are putative targets in the onset of the life-threatening HUS and could be involved in the clinical picture of the disease, whereas colon epithelial cells can be excluded as major targets for Stx1a and Stx2a.
Publications
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Real- time interaction analysis of Shiga toxins and membrane microdomains of primary human brain microvascular endothelial cells. Glycobiology.
Detzner, Johanna; Steil, Daniel; Pohlentz, Gottfried; Legros, Nadine; Humpf, Hans-Ulrich; Mellmann, Alexander; Karch, Helge & Müthing, Johannes
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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. Microorganisms, 7(11), 582.
Detzner, Johanna; Gloerfeld, Caroline; Pohlentz, Gottfried; Legros, Nadine; Humpf, Hans-Ulrich; Mellmann, Alexander; Karch, Helge & Müthing, Johannes
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Valid Presumption of Shiga Toxin-Mediated Damage of Developing Erythrocytes in EHEC-Associated Hemolytic Uremic Syndrome. Toxins, 12(6), 373.
Detzner, Johanna; Pohlentz, Gottfried & Müthing, Johannes
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Primary Human Colon Epithelial Cells (pHCoEpiCs) Do Express the Shiga Toxin (Stx) Receptor Glycosphingolipids Gb3Cer and Gb4Cer and Are Largely Refractory but Not Resistant towards Stx. International Journal of Molecular Sciences, 22(18), 10002.
Detzner, Johanna; Püttmann, Charlotte; Pohlentz, Gottfried; Humpf, Hans-Ulrich; Mellmann, Alexander; Karch, Helge & Müthing, Johannes
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Primary Human Renal Proximal Tubular Epithelial Cells (pHRPTEpiCs): Shiga Toxin (Stx) Glycosphingolipid Receptors, Stx Susceptibility, and Interaction with Membrane Microdomains. Toxins, 13(8), 529.
Detzner, Johanna; Klein, Anna-Lena; Pohlentz, Gottfried; Krojnewski, Elisabeth; Humpf, Hans-Ulrich; Mellmann, Alexander; Karch, Helge & Müthing, Johannes
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Enterohemorrhagic Escherichia coli and a Fresh View on Shiga Toxin-Binding Glycosphingolipids of Primary Human Kidney and Colon Epithelial Cells and Their Toxin Susceptibility. International Journal of Molecular Sciences, 23(13), 6884.
Detzner, Johanna; Pohlentz, Gottfried & Müthing, Johannes
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Ingenious Action of Vibrio cholerae Neuraminidase Recruiting Additional GM1 Cholera Toxin Receptors for Primary Human Colon Epithelial Cells. Microorganisms, 10(6), 1255.
Detzner, Johanna; Püttmann, Charlotte; Pohlentz, Gottfried & Müthing, Johannes
