Figure 8. Ad-Ywhaz inhibition of CIA in mice.

(A) Arthritis severity measured at 2- or 3-day intervals up to 52 days after the primary immunization with type II collagen. Intra-articular injection of 1 × 10⁸ PFU of Ad-Ywhaz or Ad-Con at 30 and 37 days are indicated by arrows (n = 8 per group and per time point). Data are representative of 2 independent experiments with similar results. (B) Ad-Ywhaz suppression of foot swelling (left, top) and chronic inflammation in the ankle joints (left, bottom), which were observed at 52 days. Mean histological severity, as assessed by the extent of inflammatory cell infiltration (IF), synovial hyperplasia (SH), and bone destruction (BD) on H&E staining, is also shown on the right. Scale bars: 200 μm. (C) Reduction of serum levels of the IgG antibody against type II collagen (anti-CII Ab) in mice injected with Ad-Ywhaz. Data are the mean ± SD arbitrary units (AU, n = 5). (D) mRNA expression levels of Ywhaz and proinflammatory cytokines (Il6 and Tnf) in the synovial tissues of CIA mice 52 days after treatment with Ad-Ywhaz or Ad-Con (n = 6 per group). Target gene expression levels were normalized to those of Gapdh (internal control). (E and F) Decrease in proinflammatory cytokine expression by Ad-Ywhaz. On day 52, lymph node (LN) or spleen cells were isolated from mice treated with Ad-Ywhaz (n = 6) or Ad-Con (n = 6), and then stimulated with 10 ng/mL LPS (E) or 1 μg/mL anti-CD3 plus anti-CD28 Abs (F) for indicated times; e.g., D2 indicates day 2. Levels of IL-6, TNF, and IL-17 in the culture supernatants were measured by ELISA. Data are the mean ± SEM for A, B, and D–F. *P < 0.05, **P < 0.01, ***P < 0.001 as determined by Student’s t test.