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

Karl E. Johansson, Anne Lie Ståhl, Ida Arvidsson, Sebastian Loos, Ashmita Tontanahal, Johan Rebetz, Milan Chromek, Ann Charlotte Kristoffersson, Ludger Johannes, Diana Karpman

Research output: Contribution to journalArticlepeer-review

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, 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.

Original languageEnglish
Article number14362
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 2019

Subject classification (UKÄ)

  • Cell and Molecular Biology

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