Ferrari D, Chiozzi P, Falzoni S, Dal Susino M, Melchiorri L, Baricordi O R, Di Virgilio F
Institute of General Pathology, University of Ferrara, Italy.
J Immunol. 1997 Aug 1;159(3):1451-8.
Extracellular ATP (ATPe) is known to cause release of processed IL-1 beta from LPS-treated macrophages and microglial cells. IL-1 beta release is fast and thought to be associated with cell death. We have reinvestigated this process to identify 1) the purinergic receptor involved; 2) the relationship to cell death; and 3) pharmacologic agonists or antagonists able to modulate IL-1 beta release. Our data confirm that ATPe is a powerful stimulus for IL-1 beta release from LPS-treated human macrophages; however, we also show that IL-1 beta release is not necessarily associated with cell death, as it occurs at lower ATP concentrations and much earlier than leakage of cytoplasmic markers. The selective purinergic P2Z receptor agonist benzoylbenzoyl ATP was at least one order of magnitude more powerful than ATP, but also had a strong cytotoxic effect. 2-Methylthio-ATP was equipotent as ATPe at the optimal concentration of 1 mM, but markedly inhibitory at higher concentrations. The irreversible P2Z blocker-oxidized ATP completely inhibited ATPe-induced IL-1 beta release. IL-1 beta release also was inhibited by increasing the K+ concentration of the incubation medium. These data suggest that ATPe triggers IL-1 beta via the purinergic P2Z receptor recently shown to be expressed by human macrophages and identified as a new member of the P2X family (P2X7), and provide pharmacologic tools for the modulation of IL-1 beta release in vitro and, possibly, in vivo.
细胞外ATP(ATPe)已知可促使经脂多糖(LPS)处理的巨噬细胞和小胶质细胞释放加工后的白细胞介素-1β(IL-1β)。IL-1β的释放迅速,且被认为与细胞死亡有关。我们重新研究了这一过程,以确定:1)所涉及的嘌呤能受体;2)与细胞死亡的关系;3)能够调节IL-1β释放的药理学激动剂或拮抗剂。我们的数据证实,ATPe是促使经LPS处理的人巨噬细胞释放IL-1β的有力刺激物;然而,我们也表明,IL-1β的释放不一定与细胞死亡相关,因为它在较低的ATP浓度下发生,且比细胞质标志物泄漏早得多。选择性嘌呤能P2Z受体激动剂苯甲酰苯甲酰ATP的效力比ATP至少高一个数量级,但也有很强的细胞毒性作用。在1 mM的最佳浓度下,2-甲硫基-ATP与ATPe的效力相当,但在较高浓度下有明显的抑制作用。不可逆的P2Z阻滞剂氧化ATP完全抑制了ATPe诱导的IL-1β释放。提高孵育培养基的钾离子浓度也抑制了IL-1β的释放。这些数据表明,ATPe通过嘌呤能P2Z受体触发IL-1β释放,最近发现该受体由人巨噬细胞表达,并被鉴定为P2X家族(P2X7)的新成员,同时提供了在体外以及可能在体内调节IL-1β释放的药理学工具。