CD4+ T cells (lanes 1 and 2) and Jurkat cells (lanes 3 and 4). Silver staining of immunoprecipitates, CD4+ T cells (lane 1) and Jurkat cells (lane 3). Immunoprecipitates analysed by Western blotting using anti-FcγRIIIA/B, lane 2 (CD4+ T cells) and lane 4 (Jurkat cells). At 29 kD a broad band with another band at 35 kD was observed in CD4+ T cells and a single band at 29 kD in Jurkat cells. Fig. S7. Immunoprecipitates obtained using anti-FcγRIIIA/B Ku-0059436 ic50 monoclonal antibodies. Proteins stained using silver (left) and blots stained with Coomassie Brilliant Blue R250. Untreated cells (lane 1), terminal complement complex

(TCC) (lane 2), TCC and immune complexes (ICs) (lane 3) and IC-treated cells (lane 4). Arrow point to a protein migrating at approximately 72 kD (Syk). Fig. S8. Inhibition of aggregated human γ-globulin (AHG) binding to CD4+ T cells by see more anti-FcγRIIIA/B monoclonal antibody. 1 × 106 cells treated with AHG-AlexaFluor®488 (5 µg), control cells treated

with isotype antibody (10 µg, left panel) and monoclonal anti-FcγRIIIA/B (10 µg, right panel). “
“Eotaxin-3/CCL26 is an agonist for chemokine receptor 3 (CCR3) and a natural antagonist for CCR1, CCR2 and CCR5. CCL26 expression by non-haematopoietic cells has been well documented; however, no studies to date have demonstrated CCL26 expression by leucocytes. In this study, we investigated the ability of human monocytic cells to produce CCL26 in response to cytokines. We found that interleukin-4 (IL-4) increased the expression of CCL26 messenger RNA (mRNA) and protein in U937 Isotretinoin cells, in human monocytes and in human monocyte-derived macrophages. Tumour necrosis factor-α (TNF-α) and interleukin-1β

(IL-1β) alone did not induce CCL26 expression, yet these pro-inflammatory cytokines synergized with IL-4 to increase CCL26 protein expression. Signal transducer and activator of transcription 6 (STAT6) was not affected by costimulation with TNF-α, suggesting that the synergy between IL-4 and TNF-α occurs at a step downstream of STAT6 activation. Co-incubation of interferon-γ (IFN-γ) with IL-4 had no effect on CCL26 protein release. By contrast, pretreatment with IFN-γ decreased total STAT6 protein, blocked IL-4-mediated STAT6 phosphorylation and decreased IL-4-mediated CCL26 mRNA expression and protein release. These data show that IL-4 and pro-inflammatory cytokines such as TNF-α, IL-1β and IFN-γ regulate CCL26 synthesis in human monocytic cells, which may be important in regulating monocyte inflammatory responses. The eotaxin subfamily of CC chemokines consists of eotaxin/CCL11, eotaxin-2/CCL24 and eotaxin-3/CCL26.1–3 Although they only share 34–39% protein homology, all eotaxins activate cells via CC chemokine receptor 3 (CCR3), which is expressed on several different cell types including eosinophils, basophils, dendritic cells, smooth muscle cells, epithelial cells and fibroblasts (reviewed in Ref. 4). CCL11 and CCL24 are expressed by haematopoietic and non-haematopoietic cells.