Valle-Reyes Salvador, Valencia-Cruz Georgina, Liñan-Rico Liliana, Pottosin Igor, Dobrovinskaya Oxana
Centro Universitario de Investigaciones Biomédicas, Universidad de Colima, Colima, Mexico.
Front Physiol. 2018 May 9;9:499. doi: 10.3389/fphys.2018.00499. eCollection 2018.
Activation of resting T cells relies on sustained Ca influx across the plasma membrane, which in turn depends on the functional expression of potassium channels, whose activity repolarizes the membrane potential. Depending on the T-cells subset, upon activation the expression of Ca- or voltage-activated K channels, KCa or Kv, is up-regulated. In this study, by means of patch-clamp technique in the whole cell mode, we have studied in detail the characteristics of Kv and KCa currents in resting and activated human T cells, the only well explored human T-leukemic cell line Jurkat, and two additional human leukemic T cell lines, CEM and MOLT-3. Voltage dependence of activation and inactivation of Kv1.3 current were shifted up to by 15 mV to more negative potentials upon a prolonged incubation in the whole cell mode and displayed little difference at a stable state in all cell lines but CEM, where the activation curve was biphasic, with a high and low potential components. In Jurkat, KCa currents were dominated by apamine-sensitive KCa2.2 channels, whereas only KCa3.1 current was detected in healthy T and leukemic CEM and MOLT-3 cells. Despite a high proliferation potential of Jurkat cells, Kv and KCa currents were unexpectedly small, more than 10-fold lesser as compared to activated healthy human T cells, CEM and MOLT-3, which displayed characteristic Kv1.3:KCa3.1 phenotype. Our results suggest that Jurkat cells represent perhaps a singular case and call for more extensive studies on primary leukemic T cell lines as well as a verification of the therapeutic potential of specific KCa3.1 blockers to combat acute lymphoblastic T leukemias.
静息T细胞的激活依赖于跨质膜的持续钙内流,而这又取决于钾通道的功能表达,钾通道的活动可使膜电位复极化。根据T细胞亚群的不同,激活后钙激活或电压激活的钾通道(KCa或Kv)的表达会上调。在本研究中,我们采用全细胞模式的膜片钳技术,详细研究了静息和激活状态下的人T细胞、唯一经过充分研究的人T白血病细胞系Jurkat以及另外两个人白血病T细胞系CEM和MOLT-3中Kv和KCa电流的特征。在全细胞模式下长时间孵育后,Kv1.3电流的激活和失活的电压依赖性向上移动了15 mV,达到更负的电位,并且在除CEM以外的所有细胞系的稳定状态下几乎没有差异,在CEM中激活曲线是双相的,有高电位和低电位成分。在Jurkat细胞中,KCa电流主要由蜂毒明肽敏感的KCa2.2通道主导,而在健康T细胞、白血病CEM和MOLT-3细胞中仅检测到KCa3.1电流。尽管Jurkat细胞具有很高的增殖潜力,但其Kv和KCa电流却出乎意料地小,与激活的健康人T细胞、CEM和MOLT-3相比小10倍以上,后者表现出特征性的Kv1.3:KCa3.1表型。我们的结果表明,Jurkat细胞可能代表了一个独特的案例,需要对原发性白血病T细胞系进行更广泛的研究,并验证特定KCa3.1阻滞剂对抗急性淋巴细胞性T白血病的治疗潜力。