Figure 3. cDC2 from COPD GOLD II lungs display increased potential to promote Tfh-like cell skewing which is associated with the increased presence of Tfh-like cells in the COPD lung.

(A) and (B) cDC2 were isolated from COPD GOLD II peritumoral lung tissues (n=7) and co-cultured with allogeneic naïve CD4⁺ T-cells. Proportions of ICOS⁺PD-1⁺ T-cells (A) and ICOS⁺PD-1⁺IL-21⁺ T-cells (B) were determined at day 7 and compared to the respective T-cell proportions induced by cDC2 from non-obstructed peritumoral lung tissues as previously shown in Figure 2 (n=10). Shown is summary data graphs in which each symbol represents an individual donor. (C) Percentages of ICOS⁺PD-1⁺ Tfh-like cells were determined in peritumoral lung tissue resections of COPD and non-obstructed control subjects via flow cytometry. Shown is summary data graph in which each symbol represents an individual donor (n=6 for controls and n=5 for COPD subjects). (D) Intracellular IL-21 staining of lung tissue ICOS⁺PD-1⁺ (purple), ICOS⁻PD-1⁺ (orange), ICOS⁺PD-1⁻ (blue) and ICOS⁻PD-1⁻ (black) T-cell subsets after PMA/ionomycin restimulation (+ Golgi-plug/Golgi-stop) in DC-free in vitro cultures. Shown are pooled data from control (full diamonds) and COPD (open diamonds) lung resections (n=11). Each symbol represents an individual donor. *p<0.05, **p<0.01, ***p<0.001, Student’s t-test (A-C) and Tukey’s multiple comparison test (D).