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. 2021 Jan 1;29(4):1512–1528. doi: 10.1016/j.ymthe.2020.12.034

Figure 3.

Figure 3

Cancer-secreted exosomes reprogram HDLECs to suppress CD8+ T cell immunity

(A) Confocal imaging showed the transfer of GFP-labeled exosomes (green) to phalloidin-labeled HDLECs (red) at the indicated time point using Transwell chamber. (B and C) Exosomes secreted by Siha and hCEp were detected by transmission electron microscopy (TEM) (B) and NanoSight analysis (C). Scale bar, 100 nm. (D) Western blot for characteristic proteins of exosomes compared with conditioned media (CM) from Siha and hCEp. (E and F) Flow cytometry analysis of PD-L1 expression (E) and tube-formation assay (F) in HDLECs treated with PBS, hCEp-exo, or Siha-exo. Scale bar, 10 μm. (G−J) Flow cytometry analysis of IFN-γ (G), CD69 (H), PD-1 (I), and Annexin V (J) expression on CD8+ T cells co-cultured with HDLECs treated with PBS, hCEp-exo, or Siha-exo. (K and L) Quantification of PD-L1 expression (K) and tube formation (L) in HDLECs with indicated treatment. (M−P) Quantification of IFN-γ (M), CD69 (N), PD-1 (O), and Annexin V (P) expression on CD8+ T cells co-cultured with HDLECs with indicated treatment. The numeric values under the western blot bands represent the protein relative expression (baseline value 1.00). Error bars represent the mean ± SD of three independent experiments. ∗∗∗p < 0.001.