Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells
Hemolytic uremic syndrome (HUS) is a rare disease primarily characterized by hemolytic anemia, thrombocytopenia, and acute renal failure. Endothelial damage is the hallmark of the pathogenesis of HUS with an infection with the Shiga toxin (Stx) producing <i>Escherichia coli</i> (STEC-HUS...
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MDPI AG
2020-07-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/12/8/483 |
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| author | Wouter J. C. Feitz Nicole C. A. J. van de Kar Ian Cheong Thea J. A. M. van der Velden Carolina G. Ortiz-Sandoval Dorothea Orth-Höller Lambert P. J. W. van den Heuvel Christoph Licht |
| author_facet | Wouter J. C. Feitz Nicole C. A. J. van de Kar Ian Cheong Thea J. A. M. van der Velden Carolina G. Ortiz-Sandoval Dorothea Orth-Höller Lambert P. J. W. van den Heuvel Christoph Licht |
| author_sort | Wouter J. C. Feitz |
| collection | DOAJ |
| description | Hemolytic uremic syndrome (HUS) is a rare disease primarily characterized by hemolytic anemia, thrombocytopenia, and acute renal failure. Endothelial damage is the hallmark of the pathogenesis of HUS with an infection with the Shiga toxin (Stx) producing <i>Escherichia coli</i> (STEC-HUS) as the main underlying cause in childhood. In this study, blood outgrowth endothelial cells (BOECs) were isolated from healthy donors serving as controls and patients recovered from STEC-HUS. We hypothesized that Stx is more cytotoxic for STEC-HUS BOECs compared to healthy donor control BOECs explained via a higher amount of Stx bound to the cell surface. Binding of Shiga toxin-2a (Stx2a) was investigated and the effect on cytotoxicity, protein synthesis, wound healing, and cell proliferation was studied in static conditions. Results show that BOECs are highly susceptible for Stx2a. Stx2a is able to bind to the cell surface of BOECs with cytotoxicity in a dose-dependent manner as a result. Pre-treatment with tumor necrosis factor alpha (TNF-α) results in enhanced Stx binding with 20–30% increased lactate dehydrogenase (LDH) release. Endothelial wound healing is delayed in a Stx2a-rich environment; however, this is not caused by an effect on the proliferation rate of BOECs. No significant differences were found between control BOECs and BOECs from recovered STEC-HUS patients in terms of Stx2a binding and inhibition of protein synthesis. |
| first_indexed | 2024-03-10T18:07:25Z |
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| institution | Directory Open Access Journal |
| issn | 2072-6651 |
| language | English |
| last_indexed | 2024-03-10T18:07:25Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj.art-5eb0a7aba7d740c988b3f38747293aad2023-11-20T08:23:16ZengMDPI AGToxins2072-66512020-07-0112848310.3390/toxins12080483Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial CellsWouter J. C. Feitz0Nicole C. A. J. van de Kar1Ian Cheong2Thea J. A. M. van der Velden3Carolina G. Ortiz-Sandoval4Dorothea Orth-Höller5Lambert P. J. W. van den Heuvel6Christoph Licht7Department of Pediatric Nephrology, Amalia Children’s Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The NetherlandsDepartment of Pediatric Nephrology, Amalia Children’s Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The NetherlandsCell Biology Program, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 1X8, CanadaDepartment of Pediatric Nephrology, Amalia Children’s Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The NetherlandsCell Biology Program, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 1X8, CanadaDivision of Hygiene and Medical Microbiology, Medical University of Innsbruck, 6020 Innsbruck, AustriaDepartment of Pediatric Nephrology, Amalia Children’s Hospital, Radboud Institute for Molecular Life Sciences, Radboudumc, 6525 GA Nijmegen, The NetherlandsCell Biology Program, Research Institute, The Hospital for Sick Children, Toronto, ON M5G 1X8, CanadaHemolytic uremic syndrome (HUS) is a rare disease primarily characterized by hemolytic anemia, thrombocytopenia, and acute renal failure. Endothelial damage is the hallmark of the pathogenesis of HUS with an infection with the Shiga toxin (Stx) producing <i>Escherichia coli</i> (STEC-HUS) as the main underlying cause in childhood. In this study, blood outgrowth endothelial cells (BOECs) were isolated from healthy donors serving as controls and patients recovered from STEC-HUS. We hypothesized that Stx is more cytotoxic for STEC-HUS BOECs compared to healthy donor control BOECs explained via a higher amount of Stx bound to the cell surface. Binding of Shiga toxin-2a (Stx2a) was investigated and the effect on cytotoxicity, protein synthesis, wound healing, and cell proliferation was studied in static conditions. Results show that BOECs are highly susceptible for Stx2a. Stx2a is able to bind to the cell surface of BOECs with cytotoxicity in a dose-dependent manner as a result. Pre-treatment with tumor necrosis factor alpha (TNF-α) results in enhanced Stx binding with 20–30% increased lactate dehydrogenase (LDH) release. Endothelial wound healing is delayed in a Stx2a-rich environment; however, this is not caused by an effect on the proliferation rate of BOECs. No significant differences were found between control BOECs and BOECs from recovered STEC-HUS patients in terms of Stx2a binding and inhibition of protein synthesis.https://www.mdpi.com/2072-6651/12/8/483hemolytic uremic syndromeshiga toxinblood outgrowth endothelial cells |
| spellingShingle | Wouter J. C. Feitz Nicole C. A. J. van de Kar Ian Cheong Thea J. A. M. van der Velden Carolina G. Ortiz-Sandoval Dorothea Orth-Höller Lambert P. J. W. van den Heuvel Christoph Licht Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells Toxins hemolytic uremic syndrome shiga toxin blood outgrowth endothelial cells |
| title | Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells |
| title_full | Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells |
| title_fullStr | Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells |
| title_full_unstemmed | Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells |
| title_short | Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells |
| title_sort | primary human derived blood outgrowth endothelial cells an appropriate in vitro model to study shiga toxin mediated damage of endothelial cells |
| topic | hemolytic uremic syndrome shiga toxin blood outgrowth endothelial cells |
| url | https://www.mdpi.com/2072-6651/12/8/483 |
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