Figure 2.

Effector function of MAGE‐A‐specific CD4⁺ and CD8⁺ T cells stimulated with overlapping 15mer peptides. IFN‐γ and TNF‐α production was assessed on both CD4⁺ and CD8⁺ T cells by intracellular cytokine staining following restimulation with overlapping peptides in a cohort of patients demonstrating a positive IFN‐γ ELISPOT (n = 10). (A) A representative flow cytometry density plot demonstrating the gating strategy used to determine the subset of IFN‐γ⁺TNF‐α⁻, IFN‐γ⁻TNF‐α⁺ and IFN‐γ⁺TNF‐α⁺ T cells following pepmix stimulation is shown. (B) The mean proportion of CD4⁺ and CD8⁺ T cells within each cytokine subset was determined and is represented as a percentage of the total CD4⁺ or CD8⁺ T‐cell pool. Error bars represent SEM. (C) The correlation between the proportion of cytokine‐secreting CD4⁺ T cells and CD8⁺ T cells. Each symbol represents an individual patient and spearman's rank correlation coefficient (r) and significance (p) are reported. (D) The proportion of CD107a‐expressing CD8⁺ T cells within subsets of IFN‐γ⁺TNF‐α⁺ and IFN‐γ⁻TNF‐α⁺ cells in response to peptide stimulation. All data excluding (A) are pooled from 10 independent experiments. Data shown as mean + SEM. Data analyzed by Mann–Whitney test, **p < 0.01.