Figure 5.

High infiltration of CCR5⁺CD66b⁺TINs in MIBC could be associated with superior pembrolizumab response. (a) Quantification analysis (left) of PD-1 expression level between CCR5⁺TIN high/low subgroups in MIBC tissue microarray (TMA) by immunohistochemistry (IHC) (n = 141). (b) Flow cytometric analysis (right) of the PD-1⁺CD8⁺T cell frequencies of CD8 T cells (n = 40) between CCR5⁺CD66b⁺TINs high/low subgroups in fresh samples of MIBC. CCR5⁺CD66b⁺TINs High/low were grouped by median. (a-b) Data were analyzed using Mann-Whiney U test, and presented as mean ± SEM. (c) Flow cytometric analysis (left) of the Annexin V⁺ PI⁺ cells frequencies and Ki67⁺ cells frequencies of CD45⁻EpCAM⁺epithelial cells (n = 24) in fresh samples treated with isotype or Pembrolizumab for 12 hours. (d) Flow cytometric analysis of the Annexin V⁺PI⁺ cells frequencies and Ki67⁺ cells frequencies of CD45⁻EpCAM⁺epithelial cells (n = 24) in CCR5⁺CD66b⁺TINs high/low subgroups. (e) Flow cytometric analysis of the IFN-γ⁺ cells frequencies of CD66b⁺TINs cells (n = 16) in fresh samples treated with isotype or pembrolizumab for 12 hours. (f) Flow cytometric analysis of the Ki67⁺, PRF-1⁺, IFN-γ⁺, GZMB⁺, IL2⁺, TNF-α⁺ cells frequencies of CD8 T cells (n = 24) in fresh samples treated with isotype or pembrolizumab for 12 hours. (d, f) CCR5⁺ CD66b⁺TINs High/low were grouped by median. (c-f) Data were analyzed using Wilcoxon signed-rank test, and presented as mean ± SEM. IFN = interferon; GZMB = granzyme B; PRF-1 = perforin; TNF = Tumor Necrosis Factor.