ArrayExpress Uploader025554FK506_anti-CD3/CD2844CsA_anti-CD3/CD2844CRACi_anti-CD3/CD2844anti-CD3/CD2844Untreated43FK506_anti-CD3/CD2833CsA_anti-CD3/CD2833CRACi_anti-CD3/CD2833anti-CD3/CD2833Untreated32FK506_anti-CD3/CD2822CRACi_anti-CD3/CD2822anti-CD3/CD2821FK506_anti-CD3/CD2811CsA_anti-CD3/CD2811CRACi_anti-CD3/CD2811anti-CD3/CD2811Untreated1arrayexpress_sid4Store operated calcium entry (SOCE) downstream of T cell receptor (TCR) activation in T lymphocytes has been shown to be mediated mainly...23357789E-GEOD-43210/profile/8773/arrayexpressuploader318cellcytokinelymphocyteDec.12, 2014Falsecharacterization-of-a-novel-crac-inhibitor-that-poE-GEOD-43210_A-AFFY-44Characterization of A Novel CRAC Inhibitor that Potently Blocks Human T cell Activation and Effector FunctionsJul.12, 2014Store operated calcium entry (SOCE) downstream of T cell receptor (TCR) activation in T lymphocytes has been shown to be mediated mainly through the Calcium Release Activated Calcium (CRAC) channel. Here, we compared the effects of a novel, potent and selective CRAC inhibitor, 2,6-Difluoro-N-{5-[4-methyl-1-(5-methyl-thiazol-2-yl)-1,2,5,6-tetrahydro-pyridin-3-yl]-pyrazin-2-yl}-benzamide (RO2959), on T cell effector functions with that of a previously reported CRAC channel inhibitor, YM-58483, and a calcineurin inhibitor Cyclosporin A (CsA). Using both electrophysiological and calcium-based fluorescence measurements, we showed that RO2959 is a potent SOCE inhibitor that blocked an IP3-dependent current in CRAC-expressing RBL-2H3 cells and CHO cells stably expressing human Orai1 and Stim1, as well as SOCE in human primary CD4+ T cells triggered by either TCR stimulation or thapsigargin treatment. Furthermore, we demonstrated that RO2959 completely inhibited cytokine production as well as T cell proliferation mediated by TCR stimulation or MLR (Mixed Lymphocyte Reaction). Lastly, we showed by gene expression array analysis that RO2959 potently blocked TCR triggered gene expression and T cell functional pathways similar to CsA and FK506. Thus, both from a functional and transcriptional level, our data provide evidence that RO2959 is a novel and selective CRAC inhibitor that potently inhibits human T cell functions. PBMC from healthy donors (n=4) were stimulated with anti-CD3/CD28 in the presence or absence of CRAC inhibitor, CsA or FK506 for 24 hrshttp://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-43210humanhttp://www.ebi.ac.uk/arrayexpress/experiments/E-GEOD-43210/samples/