Targeting gut T cell Ca2+ release-activated Ca2+ channels inhibits T cell cytokine production and T-box transcription factor T-bet in inflammatory bowel disease

Antonio Di Sabatino, Laura Rovedatti, Rejbinder Kaur, Jonathan P. Spencer, Jon T. Brown, Valerie D. Morisset, Paolo Biancheri, Nicholas A B Leakey, Jonathan I. Wilde, Laurie Scott, Gino R. Corazza, Kevin Lee, Neel Sengupta, Charles H. Knowles, Martin J. Gunthorpe, Peter G. McLean, Thomas T. MacDonald, Laurens Kruidenier

Research output: Contribution to journalArticlepeer-review

Abstract

Prolonged Ca2+ entry through Ca2+ release-activated Ca2+ (CRAC) channels is crucial in activating the Ca 2+-sensitive transcription factor NFAT, which is responsible for directing T cell proliferation and cytokine gene expression. To establish whether targeting CRAC might counteract intestinal inflammation, we evaluated the in vitro effect of a selective CRAC inhibitor on T cell cytokine production and T-bet expression by lamina propria mononuclear cells (LPMC) and biopsy specimens from inflammatory bowel disease (IBD) patients. The inhibitory activity of the CRAC blocker was investigated through patch-clamp experiments on rat basophilic leukemia cells and fluorometric imaging plate reader intracellular Ca2+ assays using thapsigargin-stimulated Jurkat T cells and its detailed selectivity profile defined using a range of in vitro radioligand binding and functional assays. Anti-CD3/CD28-stimulated LPMC and biopsy specimens from 51 patients with IBD were cultured with a range of CRAC inhibitor concentrations (0.01-10 μM). IFN-γ, IL-2, IL-8, and IL-17 were analyzed by ELISA. T-bet was determined by immunoblotting. We found that the CRAC blocker concentration-dependently inhibited CRAC current in rat basophilic leukemia cells and thapsigargin-induced Ca2+ influx in Jurkat T cells. A concentration-dependent reduction in T-bet expression and production of IFN-γ, IL-2, IL-17, but not IL-8, was observed in IBD LPMC and biopsy specimens treated with the CRAC inhibitor. In conclusion, we provide evidence that the suppression of CRAC channel function may dampen the increased T cell response in the inflamed gut, thus suggesting a promising role for CRAC inhibitor drugs in the therapeutic management of patients with IBD.

Original languageEnglish
Pages (from-to)3454-3462
Number of pages9
JournalJournal of Immunology
Volume183
Issue number5
DOIs
Publication statusPublished - Sep 1 2009

ASJC Scopus subject areas

  • Immunology
  • Medicine(all)

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