Fig. 8.

Antigen stimulation promotes an increase in the percentage of CD4⁺ KJ1-26⁺ cells that have a memory regulatory cell phenotype in tolerant mice. On day 0 BALB/c mice were injected with 2.5 × 10⁶ CD4⁺ KJ1-26⁺ cells and, on the same day, treated with OVA and the anti-CD4/anti-CD8 mAb cocktail. Mice were boosted six weeks after priming. Ten weeks post-priming, half of the mice were boosted again with OVA (closed bars, n=3) while the remaining mice were left without an additional boost (open bars, n=3). On day 75 mice were sacrificed and spleens were removed, labeled with CD4-specific mAb and KJ1-26, and either, CD44-, CD45RB-, CD62L-, CD25-, GITR-, CTLA4- and FOXP3-specific mAb. CD4⁺ KJ1-26⁺ splenocytes from untreated DO11.10 mice were also labeled with the same panel of mAbs (hatched bars, n=3). Data shows the mean ± SEM for the % CD4⁺ KJ1-26⁺ cells that express the indicated marker (panel a) and are representative of 4 experiments. * indicates statistical significance to 0.01–0.05. ** indicates statistical significance to 0.001–0.009. *** indicates statistical significance to 0.001–0.0009. The Student t test was used to determine statistical significance.