A voltage-operable current is involved in Ca2+ entry in human lymphocytes whereas I-CRAC has no apparent role

Presently, it is thought that a non-voltage-gated current is responsible for activation-induced Ca2+ entry in nonelectrically excitable cells such as lymphocytes. However, it has also been proposed that the pathway instead involves a second messenger-regulated Ca2+ channel that is voltage operable, where ''voltage operable'' is defined as an intrinsic property of the channel protein(s) rather than a requirement of normal gating. To evaluate the contribution of these currents to activation-induced Ca2+ influx, each was examined with respect to its ability to account for Ca2+ influx as reported by Ca2+-sensitive dyes. We identified a set of reagents, nordihydroguaiaretic acid and various calmodulin inhibitors, that inhibits Ca2+ entry and blocks the voltage-operable current but leaves the non-voltage-gated current unaltered. Furthermore, nordihydroguaiaretic acid inhibited Ca2+-dependent proliferation of mitogen-activated human peripheral blood mononuclear cells or Jurkat T cells and specifically blocked Ca2+-dependent interleukin 2 production by Jurkat T cells to a degree similar to the immunosuppressant drug cyclosporin A. We also identified compounds, amiloride and Mn2+, that block the non-voltage-gated current but have no effect on either the voltage-operable current or Ca2+ entry. Correspondingly, amiloride had no effect on Ca2+-dependent proliferation of Jurkat cells. These observations imply that blockade of the non-voltage-gated current does not block either Ca2+ entry or Ca2+-dependent lymphocyte proliferation, whereas blockade of the voltage-operable current does. The data suggest that the voltage-operable current may be a mediator of activation-induced Ca2+ entry in lymphocytes.