Figure 3.

Arthritic Ackr2-deficient mice have enlarged draining LNs capable of greater collagen-induced IL-17 production. Draining LNs (n=6–10) were collected from wild-type (WT) and Ackr2-deficient (Ackr2^−/−) DBA/1j mice 35 days after collagen immunization. (a) Number of LN cells retrieved. (b–g) Draining LN cells (3 × 10⁵ cells per well) were stimulated ex vivo with or without antigen (CII, 60 μg ml⁻¹). (b) Ag-induced proliferation (that is, CII minus medium alone), as determined by incorporation of tritiated thymidine ([H³]-TdR). (c–f) Concentration of IL-17, tumor necrosis factor-α (TNFα), IL-10 and GM-CSF in the medium after stimulation with antigen (CII) or medium alone (m). Significance was determined by one-way analysis of variance (ANOVA) with Bonferroni multiple comparison post tests. *P<0.05 and **P<0.01. (g) Correlation between the concentration of GM-CSF and IL-17. Significance was determined by Spearman's rho test. (h) Representative results from IL-17 intracellular flow cytometry and a bar graph showing the percentage of LN cells that were CD4⁺IL17⁺ cells. A repeat experiment yielded similar results. Significance was determined by t-test. *P<0.05.