Recruitment of CCR6-expressing Th17 cells by CCL20 secreted from plasmin-stimulated macrophages
Recruitment of CCR6-expressing Th17 cells by CCL20 secreted from plasmin-stimulated macrophages
复制标题
由纤溶酶刺激的巨噬细胞分泌的 CCL20 招募表达 CCR6 的 Th17 细胞。
DOI：
10.1093/abbs/gmt049
复制
发表时间：
2013-07-01
影响因子：
3.7
通讯作者：
Simmet, Thomas
中科院分区：
文献类型：
--
作者：
Li, Qun;Laumonnier, Yves;Simmet, Thomas
在本研究中，单核细胞来源的人巨噬细胞与棕黄色大衣区分开来。用抗CD4磁珠和AutoMACS分离系统从外周血单个核细胞中富集原始CD4T细胞，在辅助性T细胞(Th17)促进条件下极化6d，获得Th17细胞。用流式细胞仪检测Th17细胞分化频率和C-C趋化因子受体6(CCR6)的表达。逆转录-聚合酶链式反应检测纤溶酶对巨噬细胞巨噬细胞炎性蛋白-3α/C-C趋化因子配体20(CCL20)基因的诱导作用，酶联免疫吸附试验检测巨噬细胞分泌蛋白水平。用Transwell系统检测纤溶酶刺激的巨噬细胞分泌的CCL20对Th17细胞的体外趋化作用。这些结果表明，纤溶酶能刺激人单核巨噬细胞趋化因子CCL20信使RNA的表达和CCL20蛋白的释放，这一作用主要依赖于纤溶酶的蛋白分解活性和p38丝裂原活化蛋白激酶和核因子-kappaB信号通路的激活。体外培养的Th17细胞表达CCR6的细胞数为87.23~8.6。与重组人CCL20引发的趋化作用类似，纤溶酶刺激的巨噬细胞培养上清可诱导Th17细胞趋化迁移，这种迁移可被抗CCL20中和抗体抑制。这些结果表明，炎症组织中产生的纤溶酶可能诱导人巨噬细胞产生趋化因子CCL20，导致CCR6阳性的Th17细胞重新聚集到炎症部位。
In the present study, monocyte-derived human macrophages were differentiated from buffy coats. Nave CD4 T-cells enriched from peripheral blood mononuclear cells using anti-CD4 magnetic beads and the autoMACS separation system were polarized under T-helper 17 (Th17)-promoting conditions for 6 days to get Th17 cells. The frequency of Th17 cell differentiation and the expression of C-C chemokine receptor type 6 (CCR6) on Th17 cells were investigated by flow cytometry. Plasmin-triggered induction of macrophage inflammatory protein-3alpha/C-C chemokine ligand 20 (CCL20) genes in macrophages was assessed by reverse transcription-polymerase chain reaction, and secreted protein levels were measured by enzyme-linked immunosorbent assay. Th17 cell migration induced by CCL20 secreted from plasmin-stimulated macrophages was tested in vitro by chemotaxis using a transwell system. These results demonstrate that plasmin triggers the expression of chemokine CCL20 messenger RNA and the release of CCL20 protein in human monocyte-derived macrophages, which critically depend on the proteolytic activity of plasmin and activation of p38 mitogen-activated protein kinase and nuclear factor-kappaB signaling pathways. Expression of CCR6 was detected on 87.23 8.6 of Th17 cells in vitro. Similar to chemotaxis triggered by recombinant human CCL20, supernatants collected from plasmin-stimulated macrophage-induced chemotactic migration of Th17 cells, which could be inhibited by an anti-CCL20 neutralizing antibody. These results suggest that plasmin generated in inflamed tissues might elicit production of chemokine CCL20 by human macrophages leading to the recruitment of CCR6 positive Th17 cells to the inflammatory sites.