human il 17c duoset elisa  (R&D Systems)

Journal: Journal of dermatological science

Article Title: Calcipotriol and betamethasone dipropionate exert additive inhibitory effects on the cytokine expression of inflammatory dendritic cell-Th17 cell axis in psoriasis.

doi: 10.1016/j.jdermsci.2015.12.009

Figure Lengend Snippet: Fig. 2. Treatment with calcipotriol and betamethasone dipropionate affects the activity of DCs resulting in a decreased secretion of TNF-a and IL-23. Monocytes (CD14+ cells)-derived DCs were cultured in presence of IL-4 and GM-CSF for 5 days and subsequently activated by co-stimulation with LPS and IFN-g for 3 days. Cells were treated in duplicates with 100 nM calcipotriol, 1 mM betamethasone (BDP), combination of calcipotriol and betamethasone (100 nM/1 mM; ratio 1:10 as in the fixed-combination product) or a corresponding vehicle control (0.1% DMSO). Treatment was applied after the first 5 days in culture and before and throughout the 3 days of activation with LPS and IFN-g (A) or two days after activation with LPS and IFN-g and throughout the last 24 h of activation (B). Protein levels of IL-23 and TNF-a were assessed by ELISA and MSD, respectively at the end of the 3 days of activation with LPS and IFN-g. The results expressed as the percentage difference compared to vehicle treatment control are mean values SEM from independent experiments using monocytes (CD14+ cells)-derived DCs from peripheral blood of four different healthy volunteers (n = 4). One-way ANOVA followed by Tukey’s multiple comparison tests were used to compare treatment effect. ****P 0.0001; ***P 0.001; **P 0.01; *P 0.05.

Article Snippet: After 48 h of treatment, cell culture supernatants were collected for protein analysis of IL-20 (Human IL-20 Quantikine) and IL-17C (Human IL-17C DuoSet) using ELISA kits from R&D Systems and of IL-8 and IL-6 using Human ProInflammatory-4 II Ultra-Sensitive Kit from Meso Scale Diagnostics.

Techniques: Activity Assay, Derivative Assay, Cell Culture, Control, Activation Assay, Enzyme-linked Immunosorbent Assay, Comparison

Journal: Journal of dermatological science

Article Title: Calcipotriol and betamethasone dipropionate exert additive inhibitory effects on the cytokine expression of inflammatory dendritic cell-Th17 cell axis in psoriasis.

doi: 10.1016/j.jdermsci.2015.12.009

Figure Lengend Snippet: Fig. 5. Treatment with calcipotriol and betamethasone dipropionate impairs secretion of pro-inflammatory cytokines by human KCs. Human KC monolayer cultures were treated with calcipotriol (100 nM), betamethasone (1 mM), calcipotriol in combination with betamethasone (100 nM/1 mM; ratio 1:10) followed by stimulation with the mix of IL-17A, IL-22 and TNF-a for 48 h. The levels of IL-17C, IL-20 and of IL-8 and IL-6 were measured by ELISA and MSD, respectively. The results expressed as the percentage difference compared to vehicle treatment control are mean values SEM from independent experiments using human primary KCs from three (n = 3) different donors. One-way ANOVA followed by Tukey’s multiple comparison tests were used to compare treatment effect. ****P 0.0001; ***P 0.001; **P 0.01.

Article Snippet: After 48 h of treatment, cell culture supernatants were collected for protein analysis of IL-20 (Human IL-20 Quantikine) and IL-17C (Human IL-17C DuoSet) using ELISA kits from R&D Systems and of IL-8 and IL-6 using Human ProInflammatory-4 II Ultra-Sensitive Kit from Meso Scale Diagnostics.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Comparison