Fig. 4
Arthritis score, concentrations of anti-type II collagen antibody and C-telopeptide, and suppression test of supernatant from spleen cells in the experimental groups. A Sum of arthritis score. B Arthritis score on day 51. C Hind paw thickness. D Concentration of anti-chicken type II collagen antibody. E Concentration of anti-mouse type II collagen antibody. F Concentration of C-telopeptide. On day 52 or 53, spleens were harvested from all mice (n = 10/group). Splenocytes from each mouse were cultured in a 96-well plate (2 × 105 cells/well, final volume: 200 μL) with G CII, H ConA, or I LPS (100 μg/mL, 5 μg/mL, and 2.5 μg/mL respectively). Plates were incubated at 37 °C in 5% CO2. After 3 days, the BrdU assay was conducted. The stimulation index value (mean optical density of CII or mitogen-stimulated cultures/mean optical density of medium-only cultures) was calculated for each culture treatment. Data (mean ± standard error of the mean) were compared using one-way analysis of variance followed by Tukey’s multiple comparison post-hoc tests.*, Significant differences (p < 0.05) from the control (C group); #. Significant (p < 0.05) differences from the normal (N) group. CII: type II collagen, ConA: Concanavalin A, LPS: Lipopolysaccharide