Figure 5. ST and mIF analysis of immune subtypes in BrM and rGBM.

(A) Spatial expression pattern of selected marker genes: PMEL, melanoma BrM; EPCAM, lung BrM; SOX2, rGBM; MKI67, cycling cells; GFAP, brain cells. (B) Fraction of CD8-Tterm.ex among all tumor-adjacent spots in each sample. (C) Spatial distribution of CD8-Tterm.ex subtype on melanoma and lung BrM and rGBM tissue sections. (D) Box plot showing the fraction of MRC1⁺ macrophage subtypes in the neighborhood of vascular cell spots. ***P ≤ 1 × 10^–15. (E) Representative mIF images of CD3 and CD206 staining in blood vessel–enriched regions in 5 ICB-naive and 5 ICB-treated BrM tumors. Original magnification, × 20. (F) mIF quantification showing the number of CD3⁺ and CD14⁺CD206⁺ cells per mm² of tumor section within the blood vessel–enriched regions (50 μm in diameter around α-SMA⁺ vessels). The analysis includes 5 BrM and 5 BrM.ICB patients. Each dot represents a patient. For all box plots, the lower and upper bounds indicate the 25th and 75th percentiles and the middle lines the median values. P values were calculated using a 2-sided Wilcoxon’s rank-sum test.