Figure 4.

Expansion of self-antigen-specific Foxp3⁺ nTregs in peripheral lymph nodes of RelB^DCko mice. (A) Carboxyfluorescein diacetate succinimidyl ester (CFSE)-labeled CD4⁺ CD25⁺ T cells (3 × 10⁶) from wild-type mice were intravenously transferred into RelB^DCko and control mice. Eight days later cell suspensions from peripheral LN and SPL were stained for CD4 and Foxp3 and analyzed by flow cytometry. Graphs display the percentages of proliferating (CFSE^low) cells among CFSE⁺ CD4⁺ Foxp3⁺ T cells (right) as mean value + SD. Data shown are representative of two independent experiments with one mouse per genotype in each experiment. (B) PBS- or OVA peptide-loaded osmotic minipumps were subcutaneously implanted in RelB^DCko and control mice. One day later mice received an intravenous injection of 1.4 × 10⁶ CFSE-labeled CD4⁺ CD25⁺ OT-II T cells. For flow cytometric analysis, mice were sacrificed 5 days after adoptive transfer and stained for CD4 and Foxp3. Graphs display the percentages of proliferating CFSE^low cells among all transferred CFSE⁺ CD4⁺ Foxp3⁺ nTregs. Data represent the mean values + SD from one representative experiment with n = 2–5 mice per genotype and condition. (A,B) Statistical analyses were performed using unpaired student’s t-test: ns, not significant, *p < 0.05.