Integrins and cytokines activate nuclear transcription factor-κB in human neutrophils
Integrins and cytokines activate nuclear transcription factor-κB in human neutrophils
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DOI：
10.1074/jbc.m309778200
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发表时间：
2004-01-23
影响因子：
4.8
通讯作者：
Luft, FC
中科院分区：
文献类型：
--
作者：
Kettritz, R;Choi, M;Luft, FC
中性粒细胞粘附到细胞外基质对于有效的炎症反应是必要的。粘附可能通过影响细胞内信号通路来加速中性粒细胞活化。核转录因子 kappaB (NF-kappaB) 控制多种细胞功能，包括炎症、增殖和细胞存活。我们探讨了粘附在人中性粒细胞中 NF-κB 激活中的作用。用肿瘤坏死因子-α (TNF-α)、粒细胞巨噬细胞集落刺激因子 (GM-CSF)、白介素-8 (IL-8) 和甲酰基-甲硫氨酰-亮氨酰-苯丙氨酸 (fMLP) 刺激细胞。所有四种都引发中性粒细胞粘附于纤连蛋白并在纤连蛋白上扩散。 IkappaB 激酶活性测定、IkappaBalpha 降解、电迁移率测定和定量逆转录酶 PCR 表明，GM-CSF 和 IL-8 不会激活悬浮中性粒细胞中的 NF-kappaB，但会在基质贴壁条件下快速激活 NF-kappaB。相反，TNF-α 在悬浮细胞和贴壁细胞中均激活 NF-κB。 fMLP 不会激活悬浮细胞或贴壁细胞中的 NF-κB。特异性β(2)整合素阻断可防止纤连蛋白上的GM-CSF和IL-8激活NF-kappaB。与激活抗体共同刺激 CD18 和 CD11b 导致悬浮细胞中 GM-CSF 和 IL-8 激活 NF-κB。我们用细胞松弛素抑制肌动蛋白聚合，并用白皮杉醇阻断非受体激酶 Syk。两种操作均阻止了 β(2) 整合素的共刺激 NF-κB 激活信号。因此，除了β(2)整联蛋白配体结合之外，NF-κB的激活还取决于受体相关的细胞内粘着斑复合物的形成。我们得出的结论是，β(2) 整联蛋白可能提供共刺激信号，允许某些可溶性介质激活 NF-κB 通路，即使它们在悬浮液中无法激活。当人类中性粒细胞在炎症期间离开循环血液并迁移通过细胞外基质时，这种效应可能变得重要。
Neutrophil adhesion to extracellular matrix is necessary for an effective inflammatory response. Adhesion may accelerate neutrophil activation by affecting intracellular signaling pathways. The nuclear transcription factor kappaB (NF-kappaB) controls several cellular functions, including inflammation, proliferation, and cell survival. We explored the role of adhesion in NF-kappaB activation in human neutrophils. Cells were stimulated with tumor necrosis factor-alpha (TNF-alpha), granulocyte macrophage-colony-stimulating factor (GM-CSF), interleukin-8 (IL-8), and formyl-methionyl-leucyl-phenylalanine (fMLP). All four initiated neutrophil adherence to and spreading on fibronectin. GM-CSF and IL-8 did not activate NF-kappaB in suspended neutrophils but rapidly activated NF-kappaB under adherent conditions on matrix, as shown by IkappaB kinase activity assay, IkappaBalpha degradation, electromobility shift assay, and quantitative reverse transcriptase-PCR. In contrast, TNF-alpha activated NF-kappaB both in suspended cells and adherent cells. fMLP did not activate NF-kappaB in either suspended or adherent cells. Specific beta(2) integrin blockade prevented NF-kappaB activation by GM-CSF and IL-8 on fibronectin. Co-stimulating CD18 and CD11b with activating antibodies resulted in NF-kappaB activation by GM-CSF and IL-8 in suspended cells. We inhibited actin polymerization with cytochalasin and blocked the non-receptor kinase Syk with piceatannol. Both maneuvers prevented the co-stimulatory NF-kappaB-activating signal by beta(2) integrins. Thus, in addition to beta(2) integrin ligand binding, NF-kappaB activation depended on the formation of the receptor-associated intracellular focal adhesion complex. We conclude that beta(2) integrins may provide co-stimulatory signals allowing some soluble mediators to activate the NF-kappaB pathway even when they are not capable of doing so in suspension. This effect may become important when human neutrophils leave the circulating blood and migrate through extracellular matrix during inflammation.