Freedman B D, Price M A, Deutsch C J
Department of Physiology, University of Pennsylvania, Philadelphia 19104-6085.
J Immunol. 1992 Dec 15;149(12):3784-94.
The membrane potential of human PBMC was modulated in culture by isotonic high extracellular K+ (K+e), or the K+ channel blocker, charybdotoxin (ChTX), to determine the effect of depolarization on stimulated proliferation, IL-2 elaboration, and gene expression. In serum-free cultures, ChTX and high [K+]e induced a specific dose-dependent decrease in IL-2 production. ChTX inhibited proliferation of PBMC and purified T cells, decreased IL-2 elaboration 15 h after stimulation by 78.4 +/- 5.3% (n = 5), and decreased IL-2 mRNA steady-state levels by 80% between 8 and 10 h after stimulation. The IC50 for ChTX-inhibition of IL-2 elaboration and IL-2 mRNA were both 1 nM. Similarly, high [K+]e inhibited proliferation with an IC50 of 38.9 +/- 1.1 mM (n = 13), decreased IL-2 elaboration with an IC50 of 21.3 +/- 1.2 mM (n = 6), and decreased IL-2 mRNA steady-state levels with an IC50 of 18 mM. The sensitivities of both IL-2 production and proliferation to depolarization were substantially reduced by calcium, serum, and exogenous rIL-2. From these findings we conclude that membrane potential may contribute to the control of immune responsiveness in vivo.
通过等渗高细胞外钾离子(K+e)或钾离子通道阻滞剂查卡毒素(ChTX)在培养中调节人外周血单个核细胞(PBMC)的膜电位,以确定去极化对刺激增殖、白细胞介素-2(IL-2)分泌及基因表达的影响。在无血清培养中,ChTX和高[K+]e诱导IL-2产生呈特定剂量依赖性降低。ChTX抑制PBMC和纯化T细胞的增殖,刺激后15小时使IL-2分泌减少78.4±5.3%(n = 5),刺激后8至10小时使IL-2 mRNA稳态水平降低80%。ChTX抑制IL-2分泌和IL-2 mRNA的半数抑制浓度(IC50)均为1 nM。同样,高[K+]e抑制增殖的IC50为38.9±1.1 mM(n = 13),抑制IL-2分泌的IC50为21.3±1.2 mM(n = 6),降低IL-2 mRNA稳态水平的IC50为18 mM。钙、血清和外源性重组IL-2显著降低了IL-2产生和增殖对去极化的敏感性。从这些发现我们得出结论,膜电位可能在体内免疫反应性的控制中起作用。