Fig. 5.

CD8⁺ TILs from breast tumors retain polyfunctional effector cytokine production capacity for cytokines. a CD8⁺ T cells from bcPBMCs, bcTumor, bcT⁺LN, and melTumor tissues were stimulated with PMA and ionomycin for 4 h followed by intracellular staining for IFNγ, TNFα, and IL-2 production. Representative contour plots of flow cytometry data are shown. b Graph depicts calculated polyfunctionality indices for each sample type based on frequencies of non-naive CD8⁺ T cells expressing IFNγ, TNFα, and IL-2. Each symbol represents a single patient tissue sample. c SPICE graph visualizes frequencies of non-naive CD8⁺ T cells expressing IFNγ, TNFα, and IL-2 in bcPBMCs, bcTumor, melTumor with each symbol representing one patient tissue sample in each category. Summary pie graphs generated represent adjusted means for each population color-coded in the bar graph and display the phenotype of IFNγ, TNFα, and IL-2 production in those populations. Significance was calculated using one-way ANOVA and Holm–Sidak multiple comparison tests; *p < 0.05; ***p < 0.001