IFIBIO HOUSSAY   25014
INSTITUTO DE FISIOLOGIA Y BIOFISICA BERNARDO HOUSSAY
Unidad Ejecutora - UE
artículos
Título:
Comparative characterization of Shiga toxin type 2 and Subtilase cytotoxin effects on human renal epithelial and endothelial cells grown in monolayer and bilayer conditions
Autor/es:
ALVAREZ, ROMINA SOLEDAD; PATON ADRIENNE W.; AMARAL MARIA MARTA; JANCIC CAROLINA; IBARRA CRISTINA; JANCIC CAROLINA; IBARRA CRISTINA; SACERDOTI FLAVIA; PATON JAMES C.; SACERDOTI FLAVIA; PATON JAMES C.; ALVAREZ, ROMINA SOLEDAD; PATON ADRIENNE W.; AMARAL MARIA MARTA
Revista:
PLOS ONE
Editorial:
PUBLIC LIBRARY SCIENCE
Referencias:
Lugar: San Francisco; Año: 2016
ISSN:
1932-6203
Resumen:
Postdiarrheal hemolytic uremic syndrome (HUS) affects children under 5 years old and is21 responsible for the development of acute and chronic renal failure, particularly in22 Argentina. This pathology is a complication of Shiga toxin (Stx)-producing Escherichia coli23 infection and renal damage is attributed to Stx types 1 and 2 (Stx1, Stx2) produced by24 Escherichia coli O157:H7 and many other STEC serotypes. It has been reported the25 production of Subtilase cytotoxin (SubAB) by non-O157 STEC isolated from cases of26 childhood diarrhea. Therefore, it is proposed that SubAB may contribute to HUS27 pathogenesis.28 The human kidney is the most affected organ because very Stx-sensitive cells express29 high amounts of biologically active receptor. In this study, we investigated the effects of30 Stx2 and SubAB on primary cultures of human glomerular endothelial cells (HGEC) and on31 a human tubular epithelial cell line (HK-2) in monoculture and coculture conditions. We32 have established the coculture as a human renal proximal tubule model to study water33 absorption and cytotoxicity in the presence of Stx2 and SubAB.34 We obtained and characterized cocultures of HGEC and HK-2. Under basal conditions,35 HGEC monolayers exhibited the lowest electrical resistance (TEER) and the highest water36 permeability, while the HGEC/HK-2 bilayers showed the highest TEER and the lowest37 water permeability. In addition, at times as short as 20-30 minutes, Stx2 and SubAB38 caused the inhibition of water absorption across HK-2 and HGEC monolayers and this39 effect was not related to a decrease in cell viability. However, toxins did not have inhibitory40 effects on water movement across HGEC/HK-2 bilayers.41 After 72 h, Stx2 inhibited the cell viability of HGEC and HK-2 monolayers, but these effects42 were attenuated in HGEC/HK-2 bilayers. On the other hand, SubAB cytotoxicity shows a 343 tendency to be attenuated by the bilayers. Our data provide evidence about the different44 effects of these toxins on the bilayers respect to the monolayers. This in vitro model of45 communication between human renal microvascular endothelial cells and human proximal46 tubular epithelial cells is a representative model of the human proximal tubule to study the47 effects of Stx2 and SubAB related to the development of HUS.