Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/101838
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Type: Journal article
Title: Comparative characterization of Shiga toxin type 2 and subtilase cytotoxin effects on human renal epithelial and endothelial cells grown in monolayer and bilayer conditions
Author: Álvarez, R.
Sacerdoti, F.
Jancic, C.
Paton, A.
Paton, J.
Ibarra, C.
Amaral, M.
Citation: PLoS One, 2016; 11(6):e0158180-1-e0158180-14
Publisher: Public Library of Science
Issue Date: 2016
ISSN: 1932-6203
1932-6203
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Responsibility: 
Romina S. Álvarez, Flavia Sacerdoti, Carolina Jancic, Adrienne W. Paton, James C. Paton, Cristina Ibarra, María M. Amaral
Abstract: Postdiarrheal hemolytic uremic syndrome (HUS) affects children under 5 years old and is responsible for the development of acute and chronic renal failure, particularly in Argentina. This pathology is a complication of Shiga toxin (Stx)-producing Escherichia coli infection and renal damage is attributed to Stx types 1 and 2 (Stx1, Stx2) produced by Escherichia coli O157:H7 and many other STEC serotypes. It has been reported the production of Subtilase cytotoxin (SubAB) by non-O157 STEC isolated from cases of childhood diarrhea. Therefore, it is proposed that SubAB may contribute to HUS pathogenesis. The human kidney is the most affected organ because very Stx-sensitive cells express high amounts of biologically active receptor. In this study, we investigated the effects of Stx2 and SubAB on primary cultures of human glomerular endothelial cells (HGEC) and on a human tubular epithelial cell line (HK-2) in monoculture and coculture conditions. We have established the coculture as a human renal proximal tubule model to study water absorption and cytotoxicity in the presence of Stx2 and SubAB. We obtained and characterized cocultures of HGEC and HK-2. Under basal conditions, HGEC monolayers exhibited the lowest electrical resistance (TEER) and the highest water permeability, while the HGEC/HK-2 bilayers showed the highest TEER and the lowest water permeability. In addition, at times as short as 20-30 minutes, Stx2 and SubAB caused the inhibition of water absorption across HK-2 and HGEC monolayers and this effect was not related to a decrease in cell viability. However, toxins did not have inhibitory effects on water movement across HGEC/HK-2 bilayers. After 72 h, Stx2 inhibited the cell viability of HGEC and HK-2 monolayers, but these effects were attenuated in HGEC/HK-2 bilayers. On the other hand, SubAB cytotoxicity shows a tendency to be attenuated by the bilayers. Our data provide evidence about the different effects of these toxins on the bilayers respect to the monolayers. This in vitro model of communication between human renal microvascular endothelial cells and human proximal tubular epithelial cells is a representative model of the human proximal tubule to study the effects of Stx2 and SubAB related to the development of HUS.
Keywords: Kidney Glomerulus; Kidney Tubules, Proximal; Cells, Cultured; Cell Line; Endothelial Cells; Epithelial Cells; Humans; Subtilisins; Escherichia coli Proteins; Cell Culture Techniques; Cell Survival; Biological Transport; Shiga Toxin 2
Description: Published: June 23, 2016
Rights: Copyright: © 2016 Álvarez et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
RMID: 0030051342
DOI: 10.1371/journal.pone.0158180
Appears in Collections:Molecular and Biomedical Science publications

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