PMID- 27336788
OWN - NLM
STAT- MEDLINE
DCOM- 20170724
LR  - 20181113
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 6
DP  - 2016
TI  - Comparative Characterization of Shiga Toxin Type 2 and Subtilase Cytotoxin 
      Effects on Human Renal Epithelial and Endothelial Cells Grown in Monolayer and 
      Bilayer Conditions.
PG  - e0158180
LID - 10.1371/journal.pone.0158180 [doi]
LID - e0158180
AB  - 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.
FAU - Alvarez, Romina S
AU  - Alvarez RS
AD  - Laboratorio de Fisiopatogenia, Departamento de Fisiologia, Facultad de Medicina, 
      Universidad de Buenos Aires, Buenos Aires, Argentina.
FAU - Sacerdoti, Flavia
AU  - Sacerdoti F
AD  - Laboratorio de Fisiopatogenia, Departamento de Fisiologia, Facultad de Medicina, 
      Universidad de Buenos Aires, Buenos Aires, Argentina.
FAU - Jancic, Carolina
AU  - Jancic C
AD  - Laboratorio de Inmunidad Innata, Instituto de Medicina Experimental 
      (IMEX-CONICET), Academia Nacional de Medicina, Buenos Aires, Argentina.
FAU - Paton, Adrienne W
AU  - Paton AW
AD  - Research Centre for Infectious Diseases, Department of Molecular and Cellular 
      Biology, University of Adelaide, Adelaide, Australia.
FAU - Paton, James C
AU  - Paton JC
AD  - Research Centre for Infectious Diseases, Department of Molecular and Cellular 
      Biology, University of Adelaide, Adelaide, Australia.
FAU - Ibarra, Cristina
AU  - Ibarra C
AD  - Laboratorio de Fisiopatogenia, Departamento de Fisiologia, Facultad de Medicina, 
      Universidad de Buenos Aires, Buenos Aires, Argentina.
FAU - Amaral, Maria M
AU  - Amaral MM
AD  - Laboratorio de Fisiopatogenia, Departamento de Fisiologia, Facultad de Medicina, 
      Universidad de Buenos Aires, Buenos Aires, Argentina.
LA  - eng
PT  - Journal Article
DEP - 20160623
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Escherichia coli Proteins)
RN  - 0 (Shiga Toxin 2)
RN  - EC 3.4.21.- (Subtilisins)
RN  - EC 3.4.21.- (subtilase cytotoxin, E coli)
SB  - IM
MH  - Biological Transport/drug effects
MH  - Cell Culture Techniques
MH  - Cell Line
MH  - Cell Survival/drug effects
MH  - Cells, Cultured
MH  - Endothelial Cells/*drug effects/metabolism
MH  - Epithelial Cells/*drug effects/metabolism
MH  - Escherichia coli Proteins/*toxicity
MH  - Humans
MH  - Kidney Glomerulus/cytology/drug effects
MH  - Kidney Tubules, Proximal/cytology/drug effects
MH  - Shiga Toxin 2/*toxicity
MH  - Subtilisins/*toxicity
PMC - PMC4918929
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2016/06/24 06:00
MHDA- 2017/07/25 06:00
PMCR- 2016/06/23
CRDT- 2016/06/24 06:00
PHST- 2016/03/07 00:00 [received]
PHST- 2016/06/10 00:00 [accepted]
PHST- 2016/06/24 06:00 [entrez]
PHST- 2016/06/24 06:00 [pubmed]
PHST- 2017/07/25 06:00 [medline]
PHST- 2016/06/23 00:00 [pmc-release]
AID - PONE-D-16-09735 [pii]
AID - 10.1371/journal.pone.0158180 [doi]
PST - epublish
SO  - PLoS One. 2016 Jun 23;11(6):e0158180. doi: 10.1371/journal.pone.0158180. 
      eCollection 2016.