Altered T-cell phenotype in IL-10 dominant-negative receptor transgenic mice upon induction of collagen-induced arthritis. (A) Sex- and age-matched DBA/1 transgenic (Tg) and wild-type (Wt) littermate control mice were immunized with type II collagen as described in "Materials and methods." Cells from draining lymph nodes in female DBA/1 mice on day 28 postimmunization were analyzed for CD44 and CD62L expression after gating on CD4+ (left) or CD8+ T cells (right). Wt, n = 3, Tg, n = 3; *P < 0.05. (B) One representative stain of CD44, CD62L in CD3+CD4+ (left) and CD3+CD8+ (right) T cells of three repeated experiments is shown. (C) The same sources of cells described above were cultured with different concentrations of collagen for 3 days. After pulsing with 1 μCi of [3H]thymidine per well for the last 18 hours, proliferation was measured as radioactivity incorporation in counts per minute (cpm). Stimulation Index = cpm experiment/cpm control (concentration of collagen = 0). The results of one representative experiment are shown. (D) The cells were also stimulated with phorbol 12-myristate 13-acetate (50 ng/ml) and ionomycin (500 ng/ml) in the presence of BD GolgiPlug for 4 hours then stained with CD4 and intracellular IFN-γ. The statistical analysis for the percentage of IFN-γ+ cells in the CD3+CD4+ gate is shown (Wt, n = 3, Tg, n = 3; P < 0.05). One representative stain is shown (E).
Tao et al. Arthritis Research & Therapy 2011 13:R212 doi:10.1186/ar3545