Fig. 2: Prognostic implications of N20 immune cell spatial distribution in tumor core for disease-free survival (DFS) and overall survival (OS).

A The bubble chart of CD4⁺, CD8⁺, CD68⁺, and FOXP3⁺ immune cells within proximity (5, 10, 20, 30, 40, 50 μm) to tumor cells predictive pCR, DFS, and OS. B, C CD8⁺ cells and survival outcomes: Kaplan–Meier survival curves revealed a significant association between the mean N20 values of CD8⁺ cells and both DFS and OS. Higher spatial distribution of CD8+ cells corresponded to longer DFS (Undefined vs 15.90 months, P = 0.046) and OS (Undefined vs 83.10 months, P = 0.031). D, E CD68⁺ cells and OS: Lower spatial distribution of N20-CD68⁺ cells was significantly associated with longer OS (Undefined vs 76.43 months, P = 0.035) and showed a tendency for longer DFS (Undefined vs 76.43 months, P = 0.213). F, G No prognostic significance for FOXP3⁺ cells: The mean N20 values for FOXP3⁺ cells did not exhibit prognostic significance for DFS or OS. H Post-neoadjuvant therapy changes: Dynamic N20 values for CD4⁺ and CD68⁺ cells decreased significantly post-neoadjuvant therapy (0.343 vs 0.211, P = 0.003; 0.211 vs 0.109, P = 0.000). I Molecular typing: Immune cells were more densely distributed in triple-negative breast cancer (TNBC) compared to other molecular subtypes. J pCR vs non-pCR differences: N20 values for CD8⁺ immune cells differed significantly between pCR and non-pCR patients (0.371 vs 0.130, P = 0.001). N20 values for CD4⁺ cells were numerically higher in pCR patients than in non-pCR patients, although not statistically significant (0.517 vs 0.309, P = 0.073). DFS disease-free survival; OS overall survival; pCR pathological complete response; N20 average number of immune cells within 20 µm radius surrounding tumor cells in the tumor core. NC/CK the ratio of number of immune cells within 20 μm adjacent to epithelial cells over the number of tumor cells.