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Volume 169, Issue 6

PF-431396 hydrate inhibition of kinase phosphorylation during adherent-invasive infection inhibits intra-macrophage replication and inflammatory cytokine release Open Access

Abstract

Adherent-invasive (AIEC) have been implicated in the aetiology of Crohn’s disease (CD). They are characterized by an ability to adhere to and invade intestinal epithelial cells, and to replicate intracellularly in macrophages resulting in inflammation. Proline-rich tyrosine kinase 2 (PYK2) has previously been identified as a risk locus for inflammatory bowel disease and a regulator of intestinal inflammation. It is overexpressed in patients with colorectal cancer, a major long-term complication of CD. Here we show that Pyk2 levels are significantly increased during AIEC infection of murine macrophages while the inhibitor PF-431396 hydrate, which blocks Pyk2 activation, significantly decreased intramacrophage AIEC numbers. Imaging flow cytometry indicated that Pyk2 inhibition blocked intramacrophage replication of AIEC with no change in the overall number of infected cells, but a significant reduction in bacterial burden per cell. This reduction in intracellular bacteria resulted in a 20-fold decrease in tumour necrosis factor α secretion by cells post-AIEC infection. These data demonstrate a key role for Pyk2 in modulating AIEC intracellular replication and associated inflammation and may provide a new avenue for future therapeutic intervention in CD.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): AIEC , intracellular replication , Pyk2 and TNFα
Funding
This study was supported by the:
  • College of Science and Health Professions, King Saud bin Abdulaziz University for Health Sciences, Jeddah, Saudi Arabia (Award PhD studentship)
    • Principle Award Recipient: GhaithFallata
  • Biotechnology and Biological Sciences Research Council (Award BB/P003281/1)
    • Principle Award Recipient: DanielM Wall
  • Biotechnology and Biological Sciences Research Council (Award BB/K008005/1)
    • Principle Award Recipient: DanielM Wall
© 2023 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License. This article was made open access via a Publish and Read agreement between the Microbiology Society and the corresponding author’s institution.
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2023-06-13
2024-05-04
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PF-431396 hydrate inhibition of kinase phosphorylation during adherent-invasive Escherichia coli infection inhibits intra-macrophage replication and inflammatory cytokine release
Microbiology 169, 001337 (2023); https://doi.org/10.1099/mic.0.001337
/content/journal/micro/10.1099/mic.0.001337
/content/journal/micro/10.1099/mic.0.001337
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