Di Virgilio F, Bronte V, Collavo D, Zanovello P
CNR Unit, Institute of General Pathology, Padova, Italy.
J Immunol. 1989 Sep 15;143(6):1955-60.
The effects of extracellular ATP on plasma membrane permeability in mouse lymphocytes were studied with plasma membrane depolarization, uptake of ethidium bromide, and release of lactate dehydrogenase as indicators of increased permeability. Extracellular ATP induced sustained depolarization of plasma membrane potential as well as uptake of low m.w. fluorescent markers in mouse lymphocytes derived from thymus and spleen, and in two lymphoma lines YAC-1 and MBL-2. The fully ionized form ATP4- rather than MgATP2- mediated the increased permeability of the plasma membrane. Although prolonged exposure to exogenous ATP ultimately lysed the lymphocytes, two CTL populations (CHM-14 clone and CTLL-2 line) and IL-2-treated spleen lymphocytes with unrestricted killing activity were highly resistant to the permeabilizing action of extracellular ATP at all concentrations tested. In addition, CTL derived from primary immune peritoneal exudate and enriched by in vitro culture for 5 days in the presence of specific stimulator cells were also resistant to this permeabilizing effect. These findings show that exogenous ATP has a lytic effect on mouse lymphocytes but not on CTL, and suggest a role for ATP in cell-mediated cytotoxicity.
以细胞膜去极化、溴化乙锭摄取及乳酸脱氢酶释放作为通透性增加的指标,研究了细胞外ATP对小鼠淋巴细胞质膜通透性的影响。细胞外ATP可诱导来自胸腺和脾脏的小鼠淋巴细胞以及两种淋巴瘤细胞系YAC-1和MBL-2的质膜电位持续去极化,并摄取低分子量荧光标记物。完全电离形式的ATP4-而非MgATP2-介导了质膜通透性的增加。尽管长时间暴露于外源性ATP最终会使淋巴细胞裂解,但两个CTL群体(CHM-14克隆和CTLL-2细胞系)以及具有不受限制杀伤活性的经白细胞介素-2处理的脾脏淋巴细胞在所有测试浓度下对外源性ATP的通透作用均具有高度抗性。此外,源自原发性免疫腹膜渗出物并在特异性刺激细胞存在下体外培养5天富集的CTL也对这种通透作用具有抗性。这些发现表明,外源性ATP对小鼠淋巴细胞具有裂解作用,但对CTL没有作用,并提示ATP在细胞介导的细胞毒性中发挥作用。