Fig 2: Programmed cell death 1 (PD-1) and T-cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM-3) expression on CD3+ T cells in lung specimens. Hematoxylin–eosin (HE) stained 4× image (A) and 20× image (B). The square in (B) indicates the location of the immunofluorescence staining analysis. (C–H) Immunofluorescence staining of DAPI (blue), CD3 (red), PD-1 (green), TIM-3 (green), and Control IgG (green). (C,E,G) Images of 40×. (D,F,H) crop figures, respectively. Scale bars: 10 μm.

Fig 3: Correlation of the concentration of high mobility group box 1 (HMGB1) with the proportion of T-cell immunoglobulin- and mucin-domain-containing molecule-3 (TIM-3) positive T cells among CD4+ or CD8+ T cells in bronchoalveolar lavage fluid (BALF). (A) The concentration of HMGB1 in BALF was measured by ELISA. Individual patient data as well as medians and interquartile ranges are shown. (B,C) Correlation of the HMGB1 concentration with the proportion of TIM3+ cells among CD4+ T cells (B) or CD8+ T cells (C). Statistical analysis for Pearson’s correlation was performed using Student’s t-distribution. NS: not significant.

Fig 4: The associations of TIM-3 expressions with clinical outcomes in targeted agents treated patients with RCC. Representative images of kaplan-Meier survival curves of TIM-3 expression. a (PFS) and b (OS) represented TIM-3 expression either in the primary or metastatic tumor; c (PFS) and d (OS) represented TIM-3 expression in the metastatic tumor

Fig 5: Multiplex IHC analysis of TAM and TAN(A–G) Correlation between OS and M1-like TAM, M2-like TAM and the ratio of M1- and M2-like TAM in LA (A) and non-LA tumor area (B), PD-L1+ M1-like and M2-like TAM in LA (C) and non-LA tumor area (D), TAN (CD66b+ Gr) in pre-treatment biopsy (E, pre) and in post-treatment LA (F), and non-LA tumor area (G).(H) Changes in TANs between pre-treatment (pre) and matched post-treatment non-LA tumor areas (post) and their correlation with OS. Sample numbers (A–H) are the same as in Figure 2.(I–K) Correlation between CD8+PD-1+ T cells and CD4+PD-1+ T cells, as indicated, and myeloid cell densities in post-treatment tumor areas. Tumors (n = 19) subgrouped by higher versus lower density of M1-like TAM (I), M2-like TAM (J), or CD66b+ TAN (K), respectively, in LA.(L and M) Correlation between LAG3+ cells (L) and TIM3+ cells (M), respectively, and TANs in post-treatment tumor areas. Tumors subgrouped by higher versus lower density of TANs in LA.(N) mIHC images of CD66b and CXCR2, as indicated, and their overlaid image. All scale bars, 100 μm. Pseudocolors assigned by the Halo software. A representative LA shown; one square region enlarged and also shown. Arrow indicates the only CXCR2+ cell that was not CD66b+ within this region. All data shown as the mean ± SD; all comparisons made by t test; *p < 0.05; **p < 0.01; NS, not significant. See also Figure S5.