Abstract
The cytotoxic effect of Shiga-like toxin (Stx; produced by certain Escherichia coli strains) plays a central role in typical hemolytic uremic syndrome (HUS). It damages the renal endothelium by inhibiting the cellular protein synthesis. Also, the monocyte has a specific receptor for Stx but is not sensitive for the cytotoxic effect. In this work, monocytes were studied as a potential transporter for Stx to the renal endothelium. Coincubation of isolated human monocytes loaded with Stx and target cells (vero cells and human umbilical vascular endothelial cells) were performed. Transfer was determined by measuring the protein synthesis of target cells and by flow cytometry. Furthermore, the effect of a temperature shift on loaded monocytes was investigated. Stx-loaded monocytes reduced the protein synthesis of target cells. After adding an antibody against Stx, incomplete recovery occurred. Also, adding only the supernatant of coincubation was followed by protein synthesis inhibition. Stx detached from its receptor on the monocyte after a change in temperature, and no release was detected without this temperature shift. Although the monocyte plays an important role in the pathogenesis of HUS, it has no role in the transfer of Stx.
Highlights
The cytotoxic effect of Shiga-like toxin (Stx; produced by certain Escherichia coli strains) plays a central role in typical hemolytic uremic syndrome (HUS)
Stx was loaded to isolated monocytes from healthy donors and coincubated with target cells [vero cells and human umbilical cord venous endothelial cells (HUVEC)]
To investigate whether monocytes can transfer Stx2 to target cells, Stx2-loaded monocytes were added to a monolayer of these cells
Summary
The cytotoxic effect of Shiga-like toxin (Stx; produced by certain Escherichia coli strains) plays a central role in typical hemolytic uremic syndrome (HUS). It damages the renal endothelium by inhibiting the cellular protein synthesis. As the monocyte has a specific receptor, it might function as a carrier to transport Stx to the renal endothelium To investigate this hypothesis, Stx was loaded to isolated monocytes from healthy donors and coincubated with target cells [vero cells and human umbilical cord venous endothelial cells (HUVEC)]. The level of transfer was determined by measuring the protein synthesis of these target cells and the transfer of fluorescein isothiocyanate (FITC)-labeled B subunit of Stx with flow cytometry
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