il 17  (R&D Systems)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Regulatory T Cell Vaccination without Autoantigen Protects against Experimental Autoimmune Encephalomyelitis

doi: 10.4049/jimmunol.178.3.1791

Figure Lengend Snippet: Oral vaccination with Salmonella-CFAH elicits TGF-β-producing Treg cells and IL-4-, IL-10-, and IL-13-producing Teff cells in contrast to IL-17-producing, but no Th2-type cytokine-producing, Teff cells upon vaccination with Salmonella vector. Cell-sorted CD25+CD4+ and CD25−CD4+ T cells from mice orally immunized with H696 or H647 were evaluated for cytokine production following anti-CD3 and anti-CD28 costimulation. Treg cells had reduced IFN-γ production but elevated TGF-β when compared with Teff cells. IL-4, IL-10, and IL-13 segregated with the Teff cells induced by vaccination with H696. Teff cells from H647-vaccinated mice did not produce any Th2-type cytokines but rather produced elevated levels of IFN-γ. H647-induced Treg cells did produce TGF-β (although less than H696-vaccinated mice) and IL-17. Thus, protection conferred by Salmonella-CFA/I-induced Treg cells is because of reduced IFN-γ production and increased TGF-β, as well as in part supported by immune deviation by the Teff cells. *, p < 0.001 represents differences in cytokine production between CD25+CD4+ and CD25−CD4+ T cells, and †, p < 0.001 represents differences in cytokine production between H696- and H647-sorted cells.

Article Snippet: After blocking with PBS plus 1% BSA for 2 h at 37°C, washed wells were incubated with cell culture supernatants at 4°C for 24 h. After washing, 5.0 μ g/ml biotinylated chicken anti-human TGF- β 1 Ab (RD Systems) or 0.5 μ g/ml biotinylated rat anti-mouse IL-17 Ab (clone TC11-8H4; BD Pharmingen) were added for 2 h at 37°C.

Techniques: Plasmid Preparation, Produced

Journal: Scientific reports

Article Title: DJ-1 controls T cell differentiation and osteoclastogenesis in rheumatoid arthritis.

doi: 10.1038/s41598-022-16285-1

Figure Lengend Snippet: Figure 2. CD4+ T cell differentiation of peripheral blood mononuclear cells (PBMCs) determined using flow cytometry, and levels of pro-inflammatory cytokines and soluble RANKL in culture media with Th17-polarizing conditions. PBMCs from healthy controls (n = 5) were cultured under Th17 conditions (IL-1β, IL-6, IL-23 added media) with DJ-1 (10, 50, 100 ng/mL). (A) CD4+ T cell (CD4+ RANKL+ T cells, CD4+ CCR4+ CCR6+ CXCR3− T cells, CD4+ CD25high Foxp3+ T cells, and CD4+IFN-γ+ T cells).differentiation assessed using flow cytometry. (B) Levels of TNF-α, IL-17A, soluble RANKL, IL-6, IL-1β in culture media determined using ELISA. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Article Snippet: After blocking with phosphate-buffered saline (PBS)/1% bovine serum albumin /0.05% Tween 20 for 2 h at room temperature (22–25 °C), the test samples and recombinant Park7/DJ-1, TNF-α, IL-17A, RANKL, IL-6, MMP-9, VEGF (all from R&D Systems), and IL-1β (PeproTech) as standards were added to the 96-well plate and incubated at room temperature for another 2 h. The plates were washed four times with PBS and Tween 20 and then incubated with 300 ng/mL biotinylated mouse monoclonal antibodies against Park7/DJ-1, TNF-α, IL-17A, RANKL, IL-6, MMP-9, VEGF (all from R&D Systems), and IL-1β (PeproTech) for 2 h at room temperature.

Techniques: Cell Differentiation, Flow Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: Scientific reports

Article Title: DJ-1 controls T cell differentiation and osteoclastogenesis in rheumatoid arthritis.

doi: 10.1038/s41598-022-16285-1

Figure Lengend Snippet: Figure 5. Suppressive role of DJ-1 on osteoclastogenesis under IL-17A stimulation. CD14+ monocytes from HC (n = 6) were cultured with IL-17A (50 ng/mL) + M-CSF (25 ng/mL) without or with DJ-1 (10, 50, 100 ng/mL). (A) Number of TRAP+ multinucleated cells. (B) Gene expression of TRAP, OC-sTAMP, ATP6v0d2, NFATc1, and CTSK osteoclasts measured using RT-qPCR. Data were normalized to ACTB expression and are reported in relative expression units. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Scale bar = 200 μm.

Article Snippet: After blocking with phosphate-buffered saline (PBS)/1% bovine serum albumin /0.05% Tween 20 for 2 h at room temperature (22–25 °C), the test samples and recombinant Park7/DJ-1, TNF-α, IL-17A, RANKL, IL-6, MMP-9, VEGF (all from R&D Systems), and IL-1β (PeproTech) as standards were added to the 96-well plate and incubated at room temperature for another 2 h. The plates were washed four times with PBS and Tween 20 and then incubated with 300 ng/mL biotinylated mouse monoclonal antibodies against Park7/DJ-1, TNF-α, IL-17A, RANKL, IL-6, MMP-9, VEGF (all from R&D Systems), and IL-1β (PeproTech) for 2 h at room temperature.

Techniques: Cell Culture, Gene Expression, Quantitative RT-PCR, Expressing