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

Article Title: Synovial fibroblasts directly induce Th17 pathogenicity via the cyclooxygenase/prostaglandin E2 pathway, independent of IL-23.

doi: 10.4049/jimmunol.1300274

Figure Lengend Snippet: FIGURE 1. Limited contribution of IL-1b and IL-6 to the perseverance of the proinflammatory loop induced upon Th17–RASF interaction. Sorted primary memory CCR62 T cells and/or CCR6+ T cells (Th17) from healthy individuals were stimulated with anti-CD3/anti-CD28 and cultured with RASF for 2 d. (A) IL-1b, IL-6, and RORC gene transcription levels were detected by quantitative real-time PCR, and IL-6 and IL-17A protein levels were detected by ELISA in CCR62 T cell–RASF cultures (filled bars) or in Th17–RASF cultures (open bars). Gene transcription levels were normalized against HPRT values. (B) Th17–RASF cultures were stim- ulated with anti-CD3/anti-CD28 and cultured for 2 d in the presence or absence of anti–IL-1b (2.5 mg/ml) and/or anti–IL-6R (25 mg/ml). Ex- pression of indicated cytokines and MMPs was determined by ELISA in supernatant of Th17–RASF cultures in absence (black bars) or presence of anti–IL-1b (dark gray bars), anti–IL-6R (light gray bars), or both (white bars). (C) IL-23p19 and IL-12/23p40 gene transcription levels were detected by quantitative real-time PCR and normalized against HPRT gene expression values. Means and SEM are given for six healthy donors. Results are representative of at least three independent experiments. *p , 0.05, **p , 0.01, ***p , 0.001. n.d., Not detectable.

Article Snippet: IL-23 expression was detected by ELISA using the combination of biotin-conjugated anti-human IL-12/23p40 (eBioscience) and anti-human IL-23p19 (R&D Systems).

Techniques: Cell Culture, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Gene Expression

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

Article Title: IL-23 is increased in dendritic cells in multiple sclerosis and down-regulation of IL-23 by antisense oligos increases dendritic cell IL-10 production.

doi: 10.4049/jimmunol.176.12.7768

Figure Lengend Snippet: FIGURE 1. IL-23 ELISA calibrating curve. The IL-23 ELISA uses a standard approach. IL-23R/Fc Chimera is used as a capture reagent and anti-human IL-12/IL-23 p40 mAb as a detection Ab (R&D Systems). The sensitivity of the IL-23 ELISA was 78 pg/ml.

Article Snippet: For detecting Ab, 2 g/ml mouse anti-human p40 Ab (clone no. 169516; R&D Systems) was added for 2 h, followed by biotinylated rat anti-mouse mAb (Zymed Laboratories) for an additional 1 h. The sensitivity of the IL-23 ELISA was 78 pg/ml.

Techniques: Enzyme-linked Immunosorbent Assay