Schilling Tom, Gratopp Alexander, DeCoursey Thomas E, Eder Claudia
Institute of Physiology, Humboldt University, Tucholskystrasse 2, D 10117 Berlin, Germany.
J Physiol. 2002 Nov 15;545(1):93-105. doi: 10.1113/jphysiol.2002.028878.
Voltage-activated proton currents are reported for the first time in human peripheral blood T and B lymphocytes and in the human leukaemic T cell line Jurkat E6-1. The properties of H(+) currents studied using tight-seal voltage-clamp recording techniques were similar in all cells. Changing the pH gradient by one unit caused a 47 mV shift in the reversal potential, demonstrating high selectivity of the channels for protons. H(+) current activation upon membrane depolarisation had a sigmoidal time course that could be fitted by a single exponential function after a brief delay. Increasing pH(o) shifted the activation threshold to more negative potentials, and increased both the H(+) current amplitude and the rate of activation. In lymphocytes studied at pH(i) 6.0, the activation threshold was more negative and the H(+) current density was three times larger than at pH(i) 7.0. Increasing the intracellular Ca(2+) concentration to 1 microM did not change H(+) current amplitude or kinetics detectably. Extracellularly applied Zn(2+) and Cd(2+) inhibited proton currents, slowing activation and shifting the voltage-activation curve to more positive potentials. The H(+) current amplitude was 100 times larger in CD19+ B lymphocytes and in Jurkat E6-1 cells than in CD3+ T lymphocytes. Following stimulation with the phorbol ester PMA, the H(+) current density in peripheral blood T lymphocytes and Jurkat T cells increased. In contrast, the H(+) current density of phorbol ester (PMA)-stimulated B lymphocytes was reduced and activation became slower. The pattern of expression of H(+) channels in lymphocytes appears well suited to their proposed role of charge compensation during the respiratory burst.
首次在人外周血T淋巴细胞、B淋巴细胞以及人白血病T细胞系Jurkat E6 - 1中报道了电压激活质子电流。使用紧密封电压钳记录技术研究的H⁺电流特性在所有细胞中相似。将pH梯度改变一个单位会使反转电位发生47 mV的偏移,表明通道对质子具有高选择性。膜去极化时H⁺电流激活具有S形时间进程,在短暂延迟后可由单个指数函数拟合。增加细胞外pH值将激活阈值移向更负的电位,并增加了H⁺电流幅度和激活速率。在细胞内pH值为6.0时研究的淋巴细胞中,激活阈值更负,H⁺电流密度比细胞内pH值为7.0时大三倍。将细胞内Ca²⁺浓度增加到1 μM并未显著改变H⁺电流幅度或动力学。细胞外施加的Zn²⁺和Cd²⁺抑制质子电流,减慢激活并将电压激活曲线移向更正的电位。CD19⁺ B淋巴细胞和Jurkat E6 - 1细胞中的H⁺电流幅度比CD3⁺ T淋巴细胞中的大100倍。用佛波酯PMA刺激后,外周血T淋巴细胞和Jurkat T细胞中的H⁺电流密度增加。相反地,佛波酯(PMA)刺激的B淋巴细胞的H⁺电流密度降低,激活变得更慢。淋巴细胞中H⁺通道的表达模式似乎非常适合其在呼吸爆发期间电荷补偿的假定作用。