High-molecular-weight kininogen fragments stimulate the secretion of cytokines and chemokines through uPAR, Mac-1, and gC1qR in monocytes
High-molecular-weight kininogen fragments stimulate the secretion of cytokines and chemokines through uPAR, Mac-1, and gC1qR in monocytes
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DOI：
10.1161/01.atv.0000240290.70852.c0
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发表时间：
2006-10-01
影响因子：
8.7
通讯作者：
Colman, Robert W.
中科院分区：
文献类型：
--
作者：
Khan, Mohammad M.;Bradford, Harlan N.;Colman, Robert W.
目的：炎症性疾病患者血浆大分子激肽原(HK)在激肽释放酶(BK)释放过程中被激肽释放酶切割为HKA。我们假设Hka与细胞因子/趋化因子的释放有直接联系。方法和结果-Hka而不是BK从分离的人单核细胞释放细胞因子肿瘤坏死因子(TNF)-α、白介素1-β(IL-1β)、白介素6(IL-6)以及趋化因子IL-8和MCP-1。激肽原结构域3(D3)、E7P(aaG255-Q292)、HK结构域5(D5)、重组多肽HG(Aa K420-D474)和HGK(Aa H475-S626)的谷胱甘肽-S-转移酶(GST)融合蛋白在600 nM浓度下刺激单核细胞分泌IL-1β。针对D5或D3的单抗(MAb)可阻断Hka释放IL-1β，支持这两个结构域的重要性。抗白细胞表面受体Mac-1、LFA-1、uPAR和C1qR的抗体对TKA诱导的IL-1β分泌的抑制作用分别为98%、89%、85%和62%。通过对单个核细胞的分离，确定了负责的细胞--血单核细胞。信号通路抑制剂NFkB、JNK和p38而不是细胞外信号调节激酶(ERK)可减少单个核细胞释放细胞因子。结论Hka结构域3和5可能通过uPAR、β2整合素和gC1qR启动的细胞内信号通路从人单核细胞释放IL-1β，从而参与炎症性疾病的发病。
Objective - Plasma high-molecular-weight kininogen (HK) is cleaved in inflammatory diseases by kallikrein to HKa with release of bradykinin (BK). We postulated a direct link between HKa and cytokine/chemokine release.Methods and Results - HKa, but not BK, releases cytokines tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, IL-6, and chemokines IL-8 and MCP-1 from isolated human mononuclear cells. At a concentration of 600 nM, glutathione-S-transferase (GST) fusion proteins of kininogen domain 3 (D3), a fragment of domain 3, E7P (aaG255-Q292), HK domain 5 (D5), the D5 recombinant peptides HG (aa K420-D474) and HGK ( aa H475-S626) stimulated secretion of IL-1 beta from mononuclear cells. Monoclonal antibodies (MAbs) specific for D5 or specific for D3 blocked release of IL-1 beta by HKa, supporting the importance of both domains. Antibodies to HK receptors on leukocytes including Mac-1, LFA-1, uPAR, and C1qR inhibited IL-1 beta secretion induced by tKa 98%, 89%, 85%, and 62%, respectively. Fractionation of mononuclear cells identified the responsible cell, a blood monocyte. Inhibitors of signaling pathways NFkB, JNK, and p38 but not extracellular signal-regulated kinase (ERK) decreased cytokine release from mononuclear cells. HKa increased the synthesis of IL-1 beta as deduced by an increase of IL-1 beta mRNA at 1 to 2 hours.Conclusions - HKa domains 3 and 5 may contribute to the pathogenesis of inflammatory diseases by releasing IL-1 beta from human monocytes using intracellular signaling pathways initiated by uPAR, beta 2 integrins and gC1qR.