Table 3 Association detected cells per square mm within the calculated K-means clusters according to concentration of Tregs (FoxP3 +) and macrophages (CD163 +). Level of significance (p-value) was calculated using Kruskal-Wallis test
|  |  | All patient n = 101 | Cluster 1 treg high (n = 10) | Cluster 2 macrophage high (n = 16) | Cluster 3 immune low (n = 75) | p-value |
|---|---|---|---|---|---|---|
Mean cell density/mm2, (95% CI) | Double negative CD3 + T cells | 13.4 (9.1–17.8) | 33.8 (10.0–57.7) | 9.7 (5.2–14.2) | 11.5 (6.7–16.3) | 0.04 |
CD4+ T cells | 65.6 (25.7–105.5) | 112.3 (39.9–184.7) | 153.3 (0–398.8) | 40.7 (19.6–61.8) | 0.007 | |
Macrophages | 296.2 (217.3–375.1) | 519.1 (211.8–826.4) | 940.7 (721.1–1160.2) | 129.0 (86.4–171.5) |  < 0.001 | |
CD8+ T cells | 52.5 (33.6–71.4) | 149.6 (92.5–206.6) | 46.0 (1.0–91.0) | 41.0 (20.1–61.8) | 0.01 | |
PD1 + CD8 + T cells | 37.8 (19.4–56.1) | 82.4 (26.8–138.0) | 93.1 (49.2–137.1) | 20.0 (0–40.3) |  < 0.001 | |
Tregs | 21.7 (15.1–28.3) | 103.1 (76.2–130.0) | 25.9 (12.5–39.3) | 10.0 (7.1–12.9) |  < 0.001 | |
CD45 + PD-L1- immune cells | 351.1 (267.6–434.6) | 448.6 (72.9–824.2) | 205.2 (140.4–296.9) | 369.48 (266.7–472.2) | 0.8 | |
Proliferating immune cells | 46.9 (35.6–58.1) | 94.6 (25.0–164.3) | 30.1 (14.3–45.8) | 44.0 (32.1–55.9) | 0.08 | |
Proliferating tumor cells | 102.9 (63.0–142.7) | 88.0 (37.6–138.3) | 68.1 (23.7–112.5) | 112.4 (59.5–165.3) | 0.5 | |
Fibroblasts | 135.5 (105.8–165.2) | 80.6 (0–167.2) | 35.4 (21.6–49.2) | 164.5 (127.9–201.1) |  < 0.001 | |
PD-L1 + tumor cells | 41.5 (22.5–60.5) | 134.8 (24.6–244.9) | 87.4 (26.7–148.0) | 19.0 (3.0–34.9) |  < 0.001 | |
CD45 + PD-L1 + immune cells | 807 (171–1090) | 2038 (619–3126) | 1377 (711–2074) | 511 (132–701) |  < 0.001 |