[10, 12, 13] Despite their

unquestionable impact on functions of myeloid and lymphoid cells of the innate and adaptive immune system, little is known about the regulation of these important mediators by particular local conditions in specific organ systems. In the present study we aimed to get further insight into the regulation of eicosanoid metabolism by n-butyrate in human monocytes. Based on insights from a multigene signature approach evaluating a broad range of inflammation-related genes we focused here on the modulation of the expression of eicosanoid pathway-related genes after microbial activation and concomitant interference with n-butyrate. We found that in bacterially activated human monocytes activated by Toll-like receptor 2 (TLR2) and TLR4 ligation n-butyrate potentiated the expression of cyclo-oxygenase 2 (COX-2) along with increased PGE2 expression. selleckchem The implications

of these findings are discussed. RPMI-1640, supplemented with 2 mm l-glutamine, 100 μg/ml streptomycin, 100 U/ml penicillin and 10% fetal calf serum were purchased from PAA (Pasching Austria). The sodium salt of n-butyric acid, TLR4 ligand LPS from Escherichia coli 0111:B4 and TLR2 ligand Staphylococcus aureus Cowan strain A cells were purchased from Sigma (Deisenhofen, Germany). The dose of LPS used in our assays was 100 ng/ml and the n-butyrate dose was 1 mm if not indicated differently. Palbociclib supplier Human peripheral blood mononuclear cells were isolated from buffy coats (provided by the Austrian Red Cross) by density gradient centrifugation with Lymphoprep (Axis-Shield PoC AS, Oslo, Norway). Subsequently, monocytes were isolated from peripheral blood mononuclear cells by magnetic cell sorting using anti-CD14-conjugated magnetic beads purchased from Miltenyi Biotec (Bergisch-Gladbach, Germany). The purity of the monocytes was verified via FACS analysis on a FACSCalibur. Purity of isolated monocytes in all experiments was > 95% (data not shown). We here used a validated multigene signature approach to investigate transcriptional programmes triggered by n-butyrate and LPS alone or in combination.

Based on the knowledge-driven approach of innate immune cell biology and inflammatory process data mining, a signature of immunity/inflammation-associated LY294002 genes was assembled. TaqMan® array covering immunity/inflammation-related genes (pre-designed; Applied Biosystems, La Jolla, CA) were used as part of the self-designed 180-gene signature. This signature contained targets involved in immune response and inflammation, and included many upstream signalling molecules (kinases and phosphatases in hierarchical levels), transcription factors, and the downstream chemokines and cytokines. PTGS2 (also known as COX-2), a key enzyme in the biosynthesis of prostanoids, and other molecules central to eicosanoid signalling were also included on the array.