Figure 6.

Cytokine and chemokine production of T_RM and TILs. (A–C) Cytokine production by lung T_RM and TILs was determined after overnight αCD3/αCD28 stimulation. (A) The production of TNF-α and IFN-γ by CD103⁺ (top panels), CD69⁺ (middle panels), and CD69⁻ (bottom panels) tumor CD4⁺ (left panels) and CD8⁺ (right panels) TILs shown by representative contour plots (TNF-α on y-axis, IFN-γ on x-axis). (B) TNF-α⁺IFN-γ⁺ CD103⁺, CD69⁺, and CD69⁻ cells of lung and tumor CD4⁺ (top graph) and CD8⁺ (bottom graph) T cells. (C) PD-1 expression (geometric mean fluorescence intensity; GeoMFI) was quantified on cytokine⁺ (TNF-α⁺ and/or IFN-γ⁺) (black circles) and cytokine⁻ (TNF-α⁻IFN-γ⁻) (gray circles) CD103⁺, CD69⁺, and CD69⁻ lung and tumor CD4⁺ (top graph) and CD8⁺ (bottom graph) T cells. (D) The expression of CXCL13 was determined by flow cytometry in CD103⁺ (top panels), CD69⁺ (middle panels), and CD69⁻ (bottom panels) tumor CD4⁺ (left panels) and CD8⁺ (right panels) TILs and is shown by representative contour plots (PD-1 on y-axis, CXCL13 on x-axis). (E) CXCL13⁺ of CD103⁺, CD69⁺, and CD69⁻ cells was quantified in lung and tumor CD4⁺ (top graph) and CD8⁺ (bottom graph) T cells. (F) PD-1 expression (GeoMFI) was quantified on the CXCL13⁺ (black circles) and CXCL13⁻ (gray circles) CD103⁺, CD69⁺, and CD69⁻ CD4⁺ TILs. n = 4–6 paired lung-tumor samples; all adenocarcinoma. ^*p < 0.05, ^**p < 0.01, ^***p < 0.001; 2-way ANOVA with Tukey's multiple comparisons test.