Research Article

Therapeutic Treatment of Arthritic Mice with 15-Deoxy Δ12,14-Prostaglandin J2 (15d-PGJ2) Ameliorates Disease through the Suppression of Th17 Cells and the Induction of CD4+CD25FOXP3+ Cells

Figure 5

15d-PGJ2 induces regulatory T cell markers in conventional T cells. (a) FOXP3 mRNA expression was quantified by real-time PCR in draining lymph nodes from naïve mice (white bar) or collagen-immunized and challenged DBA/1 mice treated with vehicle (black bar) or 15d-PGJ2 (hatched bar) after 7 days of treatment. Results are presented as the mean ± SEM, ; compared with PBS-treated group. Total cells (2 × 106 cells/mL) from the draining lymph nodes from naïve or arthritic animals were in vitro incubated with 15d-PGJ2 (5 μM) (black bars) or vehicle (DMSO) (white bars) for 96 hours on plates coated with α-CD3. The nonadherent cells were phenotyped by flow cytometry using specific antibodies: anti-CD3 conjugated with FITC, anti-CD4 conjugated with PerCP, and anti-CD25 conjugated with APC-Cy7 (b) and anti-FOXP3 (c), anti-GITR (d), and anti-CTLA-4 (e) conjugated with PE. Lymphocytes were gated on CD4+CD25+ or CD4+CD25, and the population expressing the markers described above was subsequently analyzed. In (f), representative histograms of FOXP3, CTLA-4, and GITR are shown in each box. The values above are expressed as the mean ± SEM, which are representative of quadruplicate samples from two independent experiments (). compared with CD4+CD25 group control (vehicle).
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(f) Medium per vehicle