Recombinant human il 1β

Naïve CD4⁺ T cells were cultured in X-Vivo 15 media (Lonza, Cologne, Germany), which combines with 1% human serum and 1% penicillin–streptomycin (Both from Sigma-Aldrich, Saint Louis, USA). For the activation of the T cell receptor (TCR), cells were incubated with the antibodies from T cell Activation/Expansion Kit (Miltenyi Biotec, Bergisch Gladbach, Germany). For the induction of Th17 cells, recombinant human IL-1β (12.5 ng/μl), IL-6 (25 ng/ml; Both from Miltenyi Biotec, Bergisch Gladbach, Germany), recombinant human IL-23 (25 ng/ml; PeproTech, Rocky Hill, USA), and recombinant human TGF-β (25 ng/μl; PAN™-Biotech GmbH, Aidenbach, Germany) were added to cell culture for 96 h as described previously [29 (link)]. To ensure a high induction quality, we changed the medium and cytokines after 72 h of incubation. The induced Th17 cells were collected for quantitative real-time PCR.

Purified Zn²⁺ free, natural skin derived, 11,366-Da psoriasin peptide^{15 (link)} and recombinant human IL-1β (Miltenyi Biotec) and IL-6 (Invitrogen) were used to stimulate TERT-NHUC for 24 h. The concentrations used were 1600 nM or 5 µM, 20 ng/ml and 50 ng/ml respectively. IL-1β blocking was achieved by addition of IL-1β specific inhibitor, diacerein (Sigma, 50 µM) for at least 4 h prior to the IL-1β peptide treatment followed by overnight treatment in high glucose treated TERT-NHUC.