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Previous studies on the \neffects of channel blockers on T cell proliferation\nreported variable effectiveness due to differing experimental \nsystems. Therefore our aim was\nto investigate how the strength of the mitogenic stimulation \ninfluences the efficiency of cation channel\nblockers in inhibiting activation, cytokine secretion and \nproliferation of T cells under standardized\nconditions.\nHuman peripheral blood lymphocytes were activated via \nmonoclonal antibodies targeting the TCR-CD3\ncomplex and the co-stimulator CD28. We applied the blockers \nof Kv1.3 (Anuroctoxin), KCa3.1 (TRAM-\n34) and CRAC (2-Apb) channels of T cells either alone or in \ncombination with rapamycin, the inhibitor\nof the mammalian target of rapamycin (mTOR). Five days after \nthe stimulation ELISA and flow cytometric\nmeasurements were performed to determine IL-10 and IFN- \nsecretion, cellular viability and\nproliferation.\nOur results showed that ion channel blockers and rapamycin \ninhibit IL-10 and IFN- secretion and cell\ndivision in a dose-dependent manner. Simultaneous application \nof the blockers for each channel along\nwith rapamycin was the most effective, indicating synergy \namong the various activation pathways. Upon\nincreasing the extent of mitogenic stimulation the anti-\nproliferative effect of the ion channel blockers\ndiminished. This phenomenon may be important in understanding \nthe fine-tuning of T cell activation.\n© 2016 European Federation of Immunological Societies. \nPublished by Elsevier B.V. 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