Figure 4.

Enhanced apoptosis of TAA (vs. FluM1)-specific CD4⁺ T cells in the peripheral blood of melanoma patients. Flow cytometric analysis of CD4⁺ HLA-DR4/peptide tetramer⁺ T cells was performed as described in Figure 1. (A) Peripheral blood antigen-specific (tetramer⁺) CD4⁺ T cells from patients Mel093 and Mel096 were apoptotic status based on staining for Annexin-V and 7-AAD as monitored by multi-parameter flow cytometry (as described in Section “Materials and Methods”). Frequencies of early (Annexin-V⁺/7-AAD^neg) vs. total (Annexin-V⁺ regardless of 7-AAD status) apoptotic events is tabulated. (B) Cumulative results for total CD4⁺ T cells and antigen-specific CD4⁺ T cells are depicted in Box plot format, with heavy horizontal lines representing the mean for each group, and whisker bars representing 5th and 95th percentiles (n = 13; i.e., Mel036-Mel104). *p < 0.05 vs. FluM1 (T-test).