Flow cytometric gating strategy for analysis CD4, CD8 T cell response to Ag-pulsed bDC/MoDC. Gates were placed to define small lymphocytes (G1) and large activated lymphocytes (G2) and color coded blue to track activated lymphocytes. A third gate was placed on CD4 cells in this illustration to isolate activated cells for analysis. Visualization in forward scatter (FSC) vs CD45R0 (memory) shows activated cells that increase in size appear in G2. A mdPBMC cultured in medium alone for 6 days. B mdPBMC following Ag stimulation and culture for 6 days. Comparison of cells in A with B show only Ag specific CD4 memory T cells proliferated in response to stimulation with MMP. This population also expressed CD25, an indication of cells in this gate are activated (not shown). Only the activated proliferating cells in the upper right quadrant were selected for data analysis. Gated CD8 T cells had equivalent profiles.