Recruitment of CCR6-expressing Thl7 cells by CCL20 secreted from plasmin-stimulated macrophages  

作　　者：Qun Li Yves Laumonnier Tatiana Syrovets Thomas Simmet 

机构地区：[1]Joint Laboratory of Vascular Biology of Ruijin Hospital and Shanghai Institute of Hypertension, Shanghai Jiao Tong University School ofMedicine, Shanghai 200025, China [2]Health Science Center, Shanghai Institutes of Biological Sciences (SIBS), Chinese Academy of Sciences, Shanghai 200025, China [3]Institute of Pharmacology of Natural Products and Cllinical Pharmacology, Ulm University, D-89081 Ulm, Germany [4]Institute for Systemic Inflammation Research, University of Lübeck, 23562 Lübeck, Germany

出　　处：《Acta Biochimica et Biophysica Sinica》2013年第7期593-600,共8页生物化学与生物物理学报（英文版）

基　　金：This work was supported by the grants from the Deutsche Forschungs gemeinschaft （to T.S. and T.S.）, the Shanghai Scientific Research Innovation Program （11YZ59 to Q.L.）, the Shanghai Pujiang Program （10PJ1407300 to Q.L.）, the New Scholar Programs Foundation of Ministry of Education of China （20110073120103 to Q.L.）, and the National Natural Science Foundation of China （81202298 to Q.L.）.

摘　　要：In the present study, monocyte-derived human macro-phages were differentiated from buffy coats. Naive CD^4＋ T-cells enriched from peripheral blood mononuclear cells using anti-CD4 magnetic beads and the autoMACS separ-ation system were polarized under T-helper 17 （Thl7）-pro-moting conditions for 6 days to get Thl7 cells. The frequency of Thl7 cell differentiation and the expression of C-C chemokine receptor type 6 （CCR6） on Thl7 cells were investigated by flow cytometry. Plasmin-triggered induc-tion of macrophage inflammatory protein-3alpha/C-C che-mokine ligand 20 （CCL20） genes in macrophages was assessed by reverse transcription-polymerase chain reac-tion, and secreted protein levels were measured by enzyme-linked immunosorbent assay. Thl7 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 expres- sion of chemokine CCL20 messenger RNA and the release of CCL20 protein in human monocyte-derived macro-phages, 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 Thl7 cells in vitro. Similar to chemotaxis triggered by recombinant human CCL20, supernatants collected from plasmin-stimu- lated macrophage-induced chemotactic migration of Thl7 cells, which could be inhibited by an anti-CCL20 neutraliz-ing 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 Thl7 cells to the inflammatory sites.In the present study, monocyte-derived human macro-phages were differentiated from buffy coats. Naive CD^4＋ T-cells enriched from peripheral blood mononuclear cells using anti-CD4 magnetic beads and the autoMACS separ-ation system were polarized under T-helper 17 （Thl7）-pro-moting conditions for 6 days to get Thl7 cells. The frequency of Thl7 cell differentiation and the expression of C-C chemokine receptor type 6 （CCR6） on Thl7 cells were investigated by flow cytometry. Plasmin-triggered induc-tion of macrophage inflammatory protein-3alpha/C-C che-mokine ligand 20 （CCL20） genes in macrophages was assessed by reverse transcription-polymerase chain reac-tion, and secreted protein levels were measured by enzyme-linked immunosorbent assay. Thl7 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 expres- sion of chemokine CCL20 messenger RNA and the release of CCL20 protein in human monocyte-derived macro-phages, 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 Thl7 cells in vitro. Similar to chemotaxis triggered by recombinant human CCL20, supernatants collected from plasmin-stimu- lated macrophage-induced chemotactic migration of Thl7 cells, which could be inhibited by an anti-CCL20 neutraliz-ing 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 Thl7 cells to the inflammatory sites.

关 键 词：PLASMIN MACROPHAGES signaling CCL20 Thl 7 CCR6 

分 类 号：Q813[生物学—生物工程] TQ464.8[化学工程—制药化工]