Journal: Arthritis Research & Therapy

Article Title: Circulating levels of soluble MER in lupus reflect M2c activation of monocytes/macrophages, autoantibody specificities and disease activity

doi: 10.1186/ar4407

Figure Lengend Snippet: Soluble Mer is released by M2c monocytes/macrophages and correlates with circulating levels of soluble CD163 in systemic lupus erythematosus, and soluble CD163, like soluble Mer, correlates with Systemic Lupus Erythematosus Disease Activity Index score. Levels of soluble Mer (sMer) (A) and soluble Axl (sAxl) (B) were measured in supernatants of monocytes/macrophages cultured for 3 days in the presence of medium alone (resting M0), interferon γ (IFN-γ), granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin 17 (IL-17) (classically activated and/or proinflammatory M1 and M17), IL-4, transforming growth factor β (TGF-β), IL-10, macrophage colony-stimulating factor (M-CSF), dexamethasone (Dex), alone or in combination (alternatively activated and/or regulatory M2a and M2c cells). In 40 consecutive patients in our cohort, potential correlations between plasma levels of sMer (C) or sAxl (D) and soluble CD163 (sCD163) were examined. Levels of sCD163 were also analyzed according to circulating autoantibodies (E) and disease activity (F) . αchrom., antichromatin; SLEDAI, Systemic Lupus Erythematosus Disease Activity Index. * P < 0.05; ** P < 0.01; n.s., not significant.

Article Snippet: For sCD163 quantitation in supernatants, we used a mouse monoclonal anti-CD163 Ab (clone EDHu-1; Novus Biologicals) for capture, recombinant human CD163 (R&D Systems) for the standard curve and a biotinylated goat polyclonal anti-CD163 Ab (R&D Systems) for detection.

Techniques: Activity Assay, Cell Culture, Clinical Proteomics

Journal: Arthritis Research & Therapy

Article Title: Circulating levels of soluble MER in lupus reflect M2c activation of monocytes/macrophages, autoantibody specificities and disease activity

doi: 10.1186/ar4407

Figure Lengend Snippet: Interferon α/β is required for production of soluble Axl, and combining M2c polarizing conditions with type I interferon exposure inhibits soluble Mer production and enhances soluble Axl release. Levels of soluble Mer (sMer) (A) , soluble CD163 (sCD163) (B) and soluble Axl (sAxl) (C) were measured in supernatants of monocytes/macrophages cultured for 3 days in the presence or absence of type I interferons (IFN-α or IFN-β), macrophage growth factors (macrophage colony-stimulating factor (M-CSF) or granulocyte macrophage colony-stimulating factor (GM-CSF)), M2c polarizing agents (interleukin 10 (IL-10) or dexamethasone) or combinations of these. On day 2, low doses of lipopolysaccharide (LPS) were added for an additional 24 hours to stimulate ectodomain shedding of membrane receptors. Surface expression levels of Mer receptor tyrosine kinase (MerTK), Axl and CD163 were measured by flow cytometry (D) . Numbers shown in (D) refer to mean fluorescence intensity values. Data are representative of three independent experiments. Dex, dexamethasone. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: For sCD163 quantitation in supernatants, we used a mouse monoclonal anti-CD163 Ab (clone EDHu-1; Novus Biologicals) for capture, recombinant human CD163 (R&D Systems) for the standard curve and a biotinylated goat polyclonal anti-CD163 Ab (R&D Systems) for detection.

Techniques: Cell Culture, Membrane, Expressing, Flow Cytometry, Fluorescence