Figure 8

Presence of double-negative (DN) T cells in lymph nodes of arthritic IIJ mice. Cells were isolated from the leg-draining lymph nodes of AR and NAR littermates. They were stained and analyzed by flow cytometry. (A) Representative CD4 vs. CD8 density plots. Live CD3⁺ TCRα/β⁺ cells were gated on, and the percentage in each quadrant is shown. (B) Percentage of cells of each T-cell subset with activation/memory phenotype in arthritic mice (> 7 weeks old and arthritic for a minimum of 4 weeks) (n = 4 to 6 mice). (C) through (F) Comparison of absolute numbers of (C) cells isolated, (D) CD3⁺ TCRαβ⁺ T cells, (E) CD4⁺ CD25⁺ Foxp3⁺ T-regulatory cells and (F) single positive (CD4⁺ and CD8⁺) and DN (CD4^-CD8^-) T cells. For cell subsets, absolute numbers were back-calculated from percentages and number of total cells isolated. (G) Number of DN T cells that expressed the natural killer T-cell marker DX5 (n = 3 for each group in (C) through (G)). Data are means ± SEM. Groups were compared by performing a t-test (*P < 0.05, ***P < 0.001). Similar results were found in the spleens (data not shown).