Primary Human Derived Blood Outgrowth Endothelial Cells: An Appropriate In Vitro Model to Study Shiga Toxin Mediated Damage of Endothelial Cells

• Published in: Toxins 2020-07, Vol.12 (8), p.483
• Main Authors: Feitz, Wouter J C, van de Kar, Nicole C A J, Cheong, Ian, van der Velden, Thea J A M, Ortiz-Sandoval, Carolina G, Orth-Höller, Dorothea, van den Heuvel, Lambert P J W, Licht, Christoph
• Format: Article
• Language: English
• Subjects: Anemia; Animals; Binding; Blood; blood outgrowth endothelial cells; Cell adhesion & migration; Cell growth; Cell Proliferation; Cell surface; Cell Survival - drug effects; Cells, Cultured; Children; Chlorocebus aethiops; Cytokines; Cytotoxicity; Damage; E coli; Endothelial cells; Endothelial Cells - drug effects; Endothelium; Flow cytometry; Hemolytic anemia; Hemolytic uremic syndrome; Humans; Hypotheses; L-Lactate dehydrogenase; Lactate dehydrogenase; Lactic acid; Models, Biological; Morphology; Mortality; Pathogenesis; Pretreatment; Protein biosynthesis; Protein synthesis; Proteins; Rare diseases; Renal failure; Shiga toxin; Shiga Toxin - toxicity; Shiga-Toxigenic Escherichia coli; Studies; Thrombocytopenia; Toxicity; Toxins; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Vero Cells; Wound healing; Wound Healing - drug effects
• ISSN: 2072-6651; 2072-6651
• DOI: 10.3390/toxins12080483

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 (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.