Figure 2. Perivascular T cell–enriched regions differentiate de novo and rGBM.

A. Whole tumor quantification of CD8⁺ T cells by mIHC in de novo and rGBM. B. Representative images of the vascular region in GBM stained for hematoxylin- and eosin (top) and CD8, Foxp3 and CD68 (bottom). C. Representative heatmaps of CD8 expression in paired de novo and recurrent GBM tumors from three patients. D. Paired analysis showing percent tumor regions enriched for CD8⁺ cells among de novo and rGBM tumors. E. Schematic overview showing image analysis workflow to detect T-cell aggregates within tumor regions. F. Representative images showing CD3⁺ T cells and CD68⁺ macrophages detected among T-cell low (left) and T-cell high (right) regions in rGBM. For G, H, J, and L, multiple regions defined as T-cell high and T-cell low were identified for each de novo and rGBM sample analyzed (Supplemental Table S3). G. Quantification of CD3⁺Ki-67⁺ cells and pSTAT1⁺ cells in T-cell low and T-cell high regions in rGBM. H. Quantification of HLA-DR⁺CD68⁺ and CD68⁺pSTAT1⁺ myeloid subsets in T-cell low and T-cell high regions in rGBM. I. Images showing CD8⁺ (yellow) and FOXP3⁺ (purple) cells detected in T-cell high regions in de novo (left) and recurrent (right) GBM. J. Quantification of FOXP3⁺ cells (left) and ratio of CD8⁺ to FOXP3⁺ cells in T-cell high regions of de novo and rGBM. K. Images showing CD3⁺ (yellow), GzmB⁺ (purple), pSTAT1⁺ (brown), and CD68⁺ (teal) cells in T-cell high regions of de novo and rGBM tumors. L. Quantification of total pSTAT1 expression in T-cell high regions in de novo and rGBM tumors. Scale bars, 60 mm. *, P<0.05; **, P<0.01; ***, P<0.001; ****, P<0.0001; error bars indicate SD; statistical significance calculated using two-tailed Mann-Whitney test (A, F, G, I, K) or paired Wilcoxon test (C).