Original Article

Single Channel Properties and Regulated Expression of Ca²⁺ Release-Activated Ca²⁺ (CRAC) Channels in Human T Cells

Correspondence to: Michael D. Cahalan, Department of Physiology and Biophysics, University of California Irvine, Irvine, CA 92697-4561. Tel:(949) 824-7776 Fax:(949) 824-3143

Although the crucial role of Ca²⁺ influx in lymphocyte activation has been well documented, little is known about the properties or expression levels of Ca²⁺ channels in normal human T lymphocytes. The use of Na⁺ as the permeant ion in divalent-free solution permitted Ca²⁺ release-activated Ca²⁺ (CRAC) channel activation, kinetic properties, and functional expression levels to be investigated with single channel resolution in resting and phytohemagglutinin (PHA)-activated human T cells. Passive Ca²⁺ store depletion resulted in the opening of 41-pS CRAC channels characterized by high open probabilities, voltage-dependent block by extracellular Ca²⁺ in the micromolar range, selective Ca²⁺ permeation in the millimolar range, and inactivation that depended upon intracellular Mg²⁺ ions. The number of CRAC channels per cell increased greatly from 15 in resting T cells to 140 in activated T cells. Treatment with the phorbol ester PMA also increased CRAC channel expression to 60 channels per cell, whereas the immunosuppressive drug cyclosporin A (1 µM) suppressed the PHA-induced increase in functional channel expression. Capacitative Ca²⁺ influx induced by thapsigargin was also significantly enhanced in activated T cells. We conclude that a surprisingly low number of CRAC channels are sufficient to mediate Ca²⁺ influx in human resting T cells, and that the expression of CRAC channels increases 10-fold during activation, resulting in enhanced Ca²⁺ signaling.

Key Words: T lymphocyte, Ca²⁺ channel, CRAC channel, T cell activation, Ca²⁺ signaling

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