Enterohemorrhagic E. Coli (EHEC)—Secreted serine protease EspP stimulates electrogenic ion transport in human colonoid monolayers

Chung Ming Tse, Julie G. In, Jianyi Yin, Mark Donowitz, Michele Doucet, Jennifer Foulke-Abel, Fernando Ruiz-Perez, James P. Nataro, Nicholas Zachos, James B. Kaper, Olga N Kovbasnjuk

Research output: Contribution to journalArticle

Abstract

One of the characteristic manifestations of Shiga-toxin-producing Escherichia coli (E. coli) infection in humans, including EHEC and Enteroaggregative E. coli O104:H4, is watery diarrhea. However, neither Shiga toxin nor numerous components of the type-3 secretion system have been found to independently elicit fluid secretion. We used the adult stem-cell-derived human colonoid monolayers (HCM) to test whether EHEC-secreted extracellular serine protease P (EspP), a member of the serine protease family broadly expressed by diarrheagenic E. coli can act as an enterotoxin. We applied the Ussing chamber/voltage clamp technique to determine whether EspP stimulates electrogenic ion transport indicated by a change in short-circuit current (Isc). EspP stimulates Isc in HCM. The EspP-stimulated Isc does not require protease activity, is not cystic fibrosis transmembrane conductance regulator (CFTR)-mediated, but is partially Ca2+-dependent. EspP neutralization with a specific antibody reduces its potency in stimulating Isc. Serine Protease A, secreted by Enteroaggregative E. coli, also stimulates Isc in HCM, but this current is CFTR-dependent. In conclusion, EspP stimulates colonic CFTR-independent active ion transport and may be involved in the pathophysiology of EHEC diarrhea. Serine protease toxins from E. coli pathogens appear to serve as enterotoxins, potentially significantly contributing to watery diarrhea.

Original languageEnglish (US)
Article number351
JournalToxins
Volume10
Issue number9
DOIs
StatePublished - Sep 1 2018

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Enterohemorrhagic Escherichia coli
Ion Transport
Serine Proteases
Escherichia coli
Monolayers
Ions
Short circuit currents
Cystic Fibrosis Transmembrane Conductance Regulator
Shiga Toxin
Diarrhea
Enterotoxins
Shiga-Toxigenic Escherichia coli
Fluids and Secretions
Escherichia coli Infections
Adult Stem Cells
Clamping devices
Patch-Clamp Techniques
Pathogens
Stem cells
Peptide Hydrolases

Keywords

  • CFTR
  • Diarrhea
  • EHEC
  • Human colonoid monolayers
  • Intracellular Ca2+
  • Serine protease EspP
  • Short circuit current
  • SPATEs

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Enterohemorrhagic E. Coli (EHEC)—Secreted serine protease EspP stimulates electrogenic ion transport in human colonoid monolayers. / Tse, Chung Ming; In, Julie G.; Yin, Jianyi; Donowitz, Mark; Doucet, Michele; Foulke-Abel, Jennifer; Ruiz-Perez, Fernando; Nataro, James P.; Zachos, Nicholas; Kaper, James B.; Kovbasnjuk, Olga N.

In: Toxins, Vol. 10, No. 9, 351, 01.09.2018.

Research output: Contribution to journalArticle

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AU - Tse, Chung Ming

AU - In, Julie G.

AU - Yin, Jianyi

AU - Donowitz, Mark

AU - Doucet, Michele

AU - Foulke-Abel, Jennifer

AU - Ruiz-Perez, Fernando

AU - Nataro, James P.

AU - Zachos, Nicholas

AU - Kaper, James B.

AU - Kovbasnjuk, Olga N

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N2 - One of the characteristic manifestations of Shiga-toxin-producing Escherichia coli (E. coli) infection in humans, including EHEC and Enteroaggregative E. coli O104:H4, is watery diarrhea. However, neither Shiga toxin nor numerous components of the type-3 secretion system have been found to independently elicit fluid secretion. We used the adult stem-cell-derived human colonoid monolayers (HCM) to test whether EHEC-secreted extracellular serine protease P (EspP), a member of the serine protease family broadly expressed by diarrheagenic E. coli can act as an enterotoxin. We applied the Ussing chamber/voltage clamp technique to determine whether EspP stimulates electrogenic ion transport indicated by a change in short-circuit current (Isc). EspP stimulates Isc in HCM. The EspP-stimulated Isc does not require protease activity, is not cystic fibrosis transmembrane conductance regulator (CFTR)-mediated, but is partially Ca2+-dependent. EspP neutralization with a specific antibody reduces its potency in stimulating Isc. Serine Protease A, secreted by Enteroaggregative E. coli, also stimulates Isc in HCM, but this current is CFTR-dependent. In conclusion, EspP stimulates colonic CFTR-independent active ion transport and may be involved in the pathophysiology of EHEC diarrhea. Serine protease toxins from E. coli pathogens appear to serve as enterotoxins, potentially significantly contributing to watery diarrhea.

AB - One of the characteristic manifestations of Shiga-toxin-producing Escherichia coli (E. coli) infection in humans, including EHEC and Enteroaggregative E. coli O104:H4, is watery diarrhea. However, neither Shiga toxin nor numerous components of the type-3 secretion system have been found to independently elicit fluid secretion. We used the adult stem-cell-derived human colonoid monolayers (HCM) to test whether EHEC-secreted extracellular serine protease P (EspP), a member of the serine protease family broadly expressed by diarrheagenic E. coli can act as an enterotoxin. We applied the Ussing chamber/voltage clamp technique to determine whether EspP stimulates electrogenic ion transport indicated by a change in short-circuit current (Isc). EspP stimulates Isc in HCM. The EspP-stimulated Isc does not require protease activity, is not cystic fibrosis transmembrane conductance regulator (CFTR)-mediated, but is partially Ca2+-dependent. EspP neutralization with a specific antibody reduces its potency in stimulating Isc. Serine Protease A, secreted by Enteroaggregative E. coli, also stimulates Isc in HCM, but this current is CFTR-dependent. In conclusion, EspP stimulates colonic CFTR-independent active ion transport and may be involved in the pathophysiology of EHEC diarrhea. Serine protease toxins from E. coli pathogens appear to serve as enterotoxins, potentially significantly contributing to watery diarrhea.

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