The P2X7 Receptor and Pannexin-1 Are Both Required for the Promotion of Multinucleated Macrophages by the Inflammatory Cytokine GM-CSF
The P2X7 Receptor and Pannexin-1 Are Both Required for the Promotion of Multinucleated Macrophages by the Inflammatory Cytokine GM-CSF
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DOI：
10.4049/jimmunol.1002780
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发表时间：
2011-10-01
影响因子：
4.4
通讯作者：
Di Virgilio, Francesco
中科院分区：
文献类型：
--
作者：
Lemaire, Irma;Falzoni, Simonetta;Di Virgilio, Francesco
P2 X（7）受体（P2 X（7）R）是一种ATP门控离子通道，参与细胞间融合形成多核巨噬细胞（MA）的过程，但其在生理/病理刺激下的MA融合中的作用尚不清楚。基于以下几方面的证据，我们证明P2 X（7）R在炎症细胞因子GM-CSF诱导多核MA中是关键的：1）用氧化ATP（oATP）、KN-62和选择性拮抗剂A740003药理学抑制P2 X（7）R，可消除GM-CSF对大鼠肺泡MA和小鼠腹膜MA的作用; 2）针对P2 X（7）R功能缺陷而选择的小鼠J774 P2 X（7）低MA克隆无应答; 3）与野生型相反，来自缺乏P2 X（7）R的小鼠的MA对GM-CSF无应答。在J774 P2 X（7）高MA和大鼠肺泡MA中，GM-CSF也刺激ATP诱导的膜透化，这种作用在P2 X（7）低MA克隆中不存在，并被P2 X（7）阻断剂oATP和KN-62抑制。值得注意的是，GM-CSF对孔形成和MA融合的刺激作用均被阻断功能性Pannexin-1（Panx-1）抑制，并且GM-CSF未能刺激来自Panx-1敲除小鼠的细胞中的MA融合。我们提供了进一步的证据表明，腹膜MA的细胞外ATP释放依赖于P2 X（7），而不是Panx-1的表达，其代谢为腺苷介导P2 X（7）依赖性MA融合。这些数据表明P2 X（7）和Panx-1都是GM-CSF促进MA融合所需的，但可能通过不同的信号传导途径独立起作用。免疫学杂志，2011，187：3878-3887。
The P2X(7) receptor (P2X(7)R), an ATP-gated ion channel, has been implicated in the process of cell-to-cell fusion into multinucleated macrophages (MA), but its contribution to MA fusion driven by physiological/pathological stimuli is not clearly established. Based on several lines of evidence, we demonstrate that P2X(7)R is critical for the induction of multinucleated MA by the inflammatory cytokine GM-CSF: 1) pharmacological inhibition of P2X(7)R with oxidized ATP (oATP), KN-62, and the selective antagonist A740003 abrogated GM-CSF action on rat alveolar MA and murine peritoneal MA; 2) a murine J774 P2X(7) low MA clone, selected for defective P2X(7)R function, was unresponsive; 3) MA from mice lacking P2X(7)R failed to respond to GM-CSF, in contrast to wild-type. GM-CSF also stimulated ATP-induced membrane permeabilization in J774 P2X(7) high MA and rat alveolar MA, an effect absent in the P2X(7) low MA clone and inhibited by the P2X(7) blockers oATP and KN-62. Notably, the stimulatory effects of GM-CSF on pore formation and MA fusion were both inhibited by blocking functional Pannexin-1 (Panx-1), and GM-CSF failed to stimulate MA fusion in cells from Panx-1 knockout mice. We provide further evidence that extracellular ATP release from peritoneal MA is dependent on P2X(7) but not on Panx-1 expression and that its metabolism to adenosine mediates P2X(7)-dependent MA fusion. These data demonstrate that both P2X(7) and Panx-1 are required for GM-CSF promotion of MA fusion but likely act independently through different signaling pathway(s). The Journal of Immunology, 2011, 187: 3878-3887.