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

Di Sabatino, Antonio, Rovedatti, Laura, Kaur, Rejbinder, Spencer, Jonathan P, Brown, Jon T, Morisset, Valerie D, Biancheri, Paolo, Leakey, Nicholas A B, Wilde, Jonathan I, Scott, Laurie, Corazza, Gino R, Lee, Kevin, Sengupta, Neel, Knowles, Charles H, Gunthorpe, Martin J, McLean, Peter G, MacDonald, Thomas T and Kruidenier, Laurens (2009) 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. Journal of Immunology, 183 (5). pp. 3454-3462. ISSN 0022-1767

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Abstract

Prolonged Ca(2+) entry through Ca(2+) release-activated Ca(2+) (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 Ca(2+) 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 microM). IFN-gamma, 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 Ca(2+) influx in Jurkat T cells. A concentration-dependent reduction in T-bet expression and production of IFN-gamma, 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.

Item Type: Article
Uncontrolled Keywords: adult,aged,animals,pharmacology,metabolism,cell line, tumor,antagonists & inhibitors,humans,immunology,immunology,jurkat cells,middle aged,organ culture techniques,patch-clamp techniques,rats,antagonists & inhibitors,immunology,young adult
Faculty \ School: Faculty of Medicine and Health Sciences > Norwich Medical School
University of East Anglia > Faculty of Medicine and Health Sciences > Research Groups > Gastroenterology and Gut Biology
Depositing User: LivePure Connector
Date Deposited: 07 Aug 2018 16:30
Last Modified: 07 Aug 2018 20:31
URI: https://ueaeprints.uea.ac.uk/id/eprint/67967
DOI: 10.4049/jimmunol.0802887

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