title: Norepinephrine inhibits macrophage migration by decreasing CCR2 expression.
abstract: Increased incidences of infectious and septic complications during post-burn courses represent the main contributor to burn injury mortality. Sustained increases in catecholamine levels, especially norepinephrine (NE), contribute to immune disturbances in severely burned patients. The precise mechanisms underlying NE-mediated immunoregulation are not fully understood. Here we hypothesize that persistently elevated NE levels are associated with immunodysfunctions. We examined the effects of NE on the phenotype and functions of bone marrow-derived macrophages (BMMs). Whole mouse bone marrow cells were treated in vitro with 40 ng/mL of M-CSF and with 1 x 10(-6) M or 1 x 10(-8) M of NE or without NE for 7 days; cells were collected and stained with antibodies for CD11b, F4/80, MHC II and the inflammatory CC chemokine receptor 2 (CCR2). We found 1 x 10(-6) M of NE inhibited MHC II and CCR2 expression on CD11b(+)/F4/80(+) BMM cells. It also inhibited BMM proliferation by inhibiting CSF-1R expression. On the contrary, 1 x 10(-8) M of NE slightly increased both MHC II and CCR2 expression on CD11b(+)/F4/80(+) BMM cells but inhibited CD11b(+)/F4/80(+) BMM proliferation. MCP-1 based migration assay showed that the migration of 1 x 10(-6) M of NE-treated BMM toward MCP-1 was significantly decreased compared to BMM without NE treatment. Both 1 x 10(-8) M and 1 x 10(-6) M of NE enhanced TNF-α production and phagocytosis of FITC-Dextran. Intracellular staining of transcriptional factor MafB showed that 1 x 10(-6) M of NE treatment enhanced its expression, whereas 1 x 10(-8) M of NE decreased expression. Stimulation with LPS in the last 24-hours of BMM culture further decreased CCR2 and MHC II expression of these BMM, suggesting the synergistic effect of LPS and NE on macrophage. Our results demonstrate that NE regulates macrophage differentiation, proliferation and function, and may play a critical role in the dysfunctional immune response post-burn.
authors.affiliation: Ross Tilley Burn Centre, Sunnybrook Health Science Centre, Sunnybrook Research Institute, Division of Plastic Surgery, Department of Surgery, University of Toronto, Toronto, Ontario, Canada.
authors.forename: Fangming
authors.initials: F
authors.lastname: Xiu
authors.forename: Mile
authors.initials: M
authors.lastname: Stanojcic
authors.forename: Marc G
authors.initials: MG
authors.lastname: Jeschke
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D039481:CD11b Antigen / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D005260:Female; D005434:Flow Cytometry; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051258:MafB Transcription Factor / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009638:Norepinephrine / Q000494:pharmacology*; D010587:Phagocytosis / Q000187:drug effects; D016186:Receptor, Macrophage Colony-Stimulating Factor / Q000378:metabolism; D054390:Receptors, CCR2 / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D000943:Antigens, Differentiation; D039481:CD11b Antigen; C515335:Ccr2 protein, mouse; D000949:Histocompatibility Antigens Class II; D008070:Lipopolysaccharides; D051258:MafB Transcription Factor; C490406:Mafb protein, mouse; D054390:Receptors, CCR2; D014409:Tumor Necrosis Factor-alpha; C036763:monocyte-macrophage differentiation antigen; D016186:Receptor, Macrophage Colony-Stimulating Factor; D009638:Norepinephrine