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Shiga toxin signals via ATP and its effect is blocked by purinergic receptor antagonism

Johansson, Karl E. LU ; Ståhl, Anne Lie LU ; Arvidsson, Ida LU ; Loos, Sebastian LU ; Tontanahal, Ashmita LU ; Rebetz, Johan LU orcid ; Chromek, Milan LU ; Kristoffersson, Ann Charlotte LU ; Johannes, Ludger and Karpman, Diana LU orcid (2019) In Scientific Reports 9(1).
Abstract

Shiga toxin (Stx) is the main virulence factor of enterohemorrhagic Escherichia coli (EHEC), that cause gastrointestinal infection leading to hemolytic uremic syndrome. The aim of this study was to investigate if Stx signals via ATP and if blockade of purinergic receptors could be protective. Stx induced ATP release from HeLa cells and in a mouse model. Toxin induced rapid calcium influx into HeLa cells, as well as platelets, and a P2X1 receptor antagonist, NF449, abolished this effect. Likewise, the P2X antagonist suramin blocked calcium influx in Hela cells. NF449 did not affect toxin intracellular retrograde transport, however, cells pre-treated with NF449 exhibited significantly higher viability after exposure to Stx for 24 hours,... (More)

Shiga toxin (Stx) is the main virulence factor of enterohemorrhagic Escherichia coli (EHEC), that cause gastrointestinal infection leading to hemolytic uremic syndrome. The aim of this study was to investigate if Stx signals via ATP and if blockade of purinergic receptors could be protective. Stx induced ATP release from HeLa cells and in a mouse model. Toxin induced rapid calcium influx into HeLa cells, as well as platelets, and a P2X1 receptor antagonist, NF449, abolished this effect. Likewise, the P2X antagonist suramin blocked calcium influx in Hela cells. NF449 did not affect toxin intracellular retrograde transport, however, cells pre-treated with NF449 exhibited significantly higher viability after exposure to Stx for 24 hours, compared to untreated cells. NF449 protected HeLa cells from protein synthesis inhibition and from Stx-induced apoptosis, assayed by caspase 3/7 activity. The latter effect was confirmed by P2X1 receptor silencing. Stx induced the release of toxin-positive HeLa cell- and platelet-derived microvesicles, detected by flow cytometry, an effect significantly reduced by NF449 or suramin. Suramin decreased microvesicle levels in mice injected with Stx or inoculated with Stx-producing EHEC. Taken together, we describe a novel mechanism of Stx-mediated cellular injury associated with ATP signaling and inhibited by P2X receptor blockade.

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author
; ; ; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Scientific Reports
volume
9
issue
1
article number
14362
publisher
Nature Publishing Group
external identifiers
  • scopus:85073079411
  • pmid:31591425
ISSN
2045-2322
DOI
10.1038/s41598-019-50692-1
language
English
LU publication?
yes
id
9c405239-be69-4c65-ae63-10162725666a
date added to LUP
2019-10-22 09:52:09
date last changed
2024-04-16 21:54:11
@article{9c405239-be69-4c65-ae63-10162725666a,
  abstract     = {{<p>Shiga toxin (Stx) is the main virulence factor of enterohemorrhagic Escherichia coli (EHEC), that cause gastrointestinal infection leading to hemolytic uremic syndrome. The aim of this study was to investigate if Stx signals via ATP and if blockade of purinergic receptors could be protective. Stx induced ATP release from HeLa cells and in a mouse model. Toxin induced rapid calcium influx into HeLa cells, as well as platelets, and a P2X1 receptor antagonist, NF449, abolished this effect. Likewise, the P2X antagonist suramin blocked calcium influx in Hela cells. NF449 did not affect toxin intracellular retrograde transport, however, cells pre-treated with NF449 exhibited significantly higher viability after exposure to Stx for 24 hours, compared to untreated cells. NF449 protected HeLa cells from protein synthesis inhibition and from Stx-induced apoptosis, assayed by caspase 3/7 activity. The latter effect was confirmed by P2X1 receptor silencing. Stx induced the release of toxin-positive HeLa cell- and platelet-derived microvesicles, detected by flow cytometry, an effect significantly reduced by NF449 or suramin. Suramin decreased microvesicle levels in mice injected with Stx or inoculated with Stx-producing EHEC. Taken together, we describe a novel mechanism of Stx-mediated cellular injury associated with ATP signaling and inhibited by P2X receptor blockade.</p>}},
  author       = {{Johansson, Karl E. and Ståhl, Anne Lie and Arvidsson, Ida and Loos, Sebastian and Tontanahal, Ashmita and Rebetz, Johan and Chromek, Milan and Kristoffersson, Ann Charlotte and Johannes, Ludger and Karpman, Diana}},
  issn         = {{2045-2322}},
  language     = {{eng}},
  number       = {{1}},
  publisher    = {{Nature Publishing Group}},
  series       = {{Scientific Reports}},
  title        = {{Shiga toxin signals via ATP and its effect is blocked by purinergic receptor antagonism}},
  url          = {{http://dx.doi.org/10.1038/s41598-019-50692-1}},
  doi          = {{10.1038/s41598-019-50692-1}},
  volume       = {{9}},
  year         = {{2019}},
}