Flow cytometry analysis of CD4⁺ T cells stimulated with dendritic cells presenting Ep63K peptide.

<p>Expression of Foxp3^GFP and CD25 is shown on gated CD4⁺ T cells. (A) Analysis of the CD4⁺ T cells from control (left) and draining (middle) lymph nodes of TCR^mini-Foxp3^GFP mice immunized with dendritic cells presenting Ep63K and CD4⁺ T cells isolated from TCR^mini-Foxp3^GFP mice and stimulated with dendritic cells <i>in vitro</i> (right). Dendritic cells (2×10⁵) were injected into footpads of TCR^mini-Foxp3^GFP mice for three days and mice were sacrificed after 5 days and popliteal lymph nodes were analyzed. (B) Analysis of gated CD4⁺ T cells isolated from tumors of tumor-bearing TCR^mini-Foxp3^GFP control mice (left) and mice immunized with dendritic cells (middle). TCR^mini-Foxp3^GFP mice were inoculated with B16 melanoma cells expressing NP-EP63K and at the same time dendritic cells were injected (5×10⁴). Injections of dendritic cells continued daily until mice were sacrificed. The abundance (%) of CD4⁺ T cells subsets CD25^-Foxp^GFP+, CD25⁺Foxp^GFP+ and CD25⁺Foxp^GFP- in the tumor tissue of TCR^mini-Foxp3^GFP control (dots) and immunized (stripes) mice. At least three mice were analyzed in each group.