Journal: Respiratory Research

Article Title: Th17 cytokines induce pro-fibrotic cytokines release from human eosinophils

doi: 10.1186/1465-9921-14-34

Figure Lengend Snippet: IL-17 and IL-23 enhance eosinophil expression of pro-fibrotic cytokines. ( A ) Surface expression of IL-17R on eosinophils (1×10 6 cells) isolated from healthy and asthmatics was determined by flow cytometry. Blots are representative data for eosinophils isolated from one healthy control and one asthmatic patient. The graph shows arithmetic mean ± SD of IL-17R positive eosinophils as percentage of total eosinophils (n = 5). 2×10 6 peripheral blood eosinophils isolated from 10 asthmatic and 10 controls subjects were stimulated with IL-17A, F, and IL-23 (50 ng/ml or 25 ng/ml) alone or in combination for 4 hrs. Total RNA was extracted and mRNA levels of TGF-β and IL-11 were then quantified using real-time PCR. mRNA expression levels of TGF-β ( B ) and IL-11 ( C ) were normalized with GAPDH for asthmatic versus healthy individuals.

Article Snippet: Eosinophils (2×10 6 cells/ml cultured in 24 well plate) were stimulated with Th1 (IL-2, IFN-γ) (50 ng/ml), Th2 (IL-4, IL-5, IL-9, IL-13) (50 ng/ml), and Th17 (IL-17A, IL-17 F, IL-23) (10, 25, 50, or 100 ng/ml) cytokines (R&D Systems, Minneapolis, Minn., USA) for 24 hrs and supernatants were collected.

Techniques: Expressing, Isolation, Flow Cytometry, Real-time Polymerase Chain Reaction

Journal: Respiratory Research

Article Title: Th17 cytokines induce pro-fibrotic cytokines release from human eosinophils

doi: 10.1186/1465-9921-14-34

Figure Lengend Snippet: Th17 cytokines enhance eosinophil production and release of pro-fibrotic cytokines. Levels of TGF-β ( A ) and IL-11 ( B ) in the supernatant of stimulated eosinophils (1×10 6 cells/0.5 ml) were determined 24 hrs following Th17 cytokine stimulation (0-100 ng/ml) using ELISA assay. Results are expressed as the arithmetic mean ± SD from 5 independent experiments. * = p < 0.05.

Article Snippet: Eosinophils (2×10 6 cells/ml cultured in 24 well plate) were stimulated with Th1 (IL-2, IFN-γ) (50 ng/ml), Th2 (IL-4, IL-5, IL-9, IL-13) (50 ng/ml), and Th17 (IL-17A, IL-17 F, IL-23) (10, 25, 50, or 100 ng/ml) cytokines (R&D Systems, Minneapolis, Minn., USA) for 24 hrs and supernatants were collected.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: Respiratory Research

Article Title: Th17 cytokines induce pro-fibrotic cytokines release from human eosinophils

doi: 10.1186/1465-9921-14-34

Figure Lengend Snippet: P38 MAP Kinase activation is required for IL-17 enhancement of eosinophil derived pro-fibrotic cytokines. Eosinophils were isolated from peripheral blood of 10 asthmatic patients and 2×10 6 /ml cells were treated, or not, with p38 MAPK or PI3K inhibitors (SB2035802 and PI103, respectively) 2 hours prior to stimulation with IL-17 (50 ng/ml). Levels of TGF-β ( A ) and IL-11 ( B ) in the supernatant of stimulated eosinophils were then determined 24 hrs following Th17 cytokine stimulation using ELISA assay (n = 10). ( C ) Induction of p38 MAPK phosphorylation by a combination of IL-17A and IL-17 F (50 ng/ml each) is detected by western analysis. The western data shown represent one of similar results from 4 independent experiments. * = p < 0.05 compared to non-stimulated (NS). ** = p < 0.05 compared to stimulated not inhibited.

Article Snippet: Eosinophils (2×10 6 cells/ml cultured in 24 well plate) were stimulated with Th1 (IL-2, IFN-γ) (50 ng/ml), Th2 (IL-4, IL-5, IL-9, IL-13) (50 ng/ml), and Th17 (IL-17A, IL-17 F, IL-23) (10, 25, 50, or 100 ng/ml) cytokines (R&D Systems, Minneapolis, Minn., USA) for 24 hrs and supernatants were collected.

Techniques: Activation Assay, Derivative Assay, Isolation, Enzyme-linked Immunosorbent Assay, Western Blot