Figure 3

Stimulation of antigen-specific CTLs by CXCR3⁺CD4⁺/DCs. Freshly isolated CD8⁺ T cells were weekly stimulated with autologous DCs pulsed with WT-1 peptide, as described in Methods. (A) The total viable cell count for thrice (21 days for WT-1) stimulation culture was determined by trypan blue dye exclusion. The data is a representative of three independent experiments. An asterisk indicates that the number of spots obtained when CTLs were generated with CXCR3⁺CD4⁺/DC was significantly higher than the mDC and CD4⁺/DC (^*P = 0.031). (B) The CTLs (as responders) were tested against peptide or empty loaded LCLs as stimulators at a ratio of 10:1. Spots were counted by computer-assisted image analysis. Asterisks indicate that the number of spots obtained when CTLs were generated with CXCR3⁺CD4⁺/DC was significantly higher (^*P = 0.05 compared to mDC; ^**P = 0.0092 compared to CD4⁺/DC). A representative IFN-γ ELISPOT experiment of three performed is shown.