Tumor-associated macrophages and Tregs influence and represent immune cell infiltration of muscle-invasive bladder cancer and predict prognosis

Table 2 Cox-Regression model for overall survival (OS).

		Hazard ratio (univariate)	p-value	Hazard ratio (multivariate)	p-value
Gender	Female vs. male	1.3 (0.7–2.3)	0.43
Tumor stage	pT3 vs. pT2	2.2 (1.1–4.5)	0.02	2.47 (1.2–5-1)	0.01
Tumor stage	pT4 vs. pT2	4.1 (1.8–9.0)	< 0.001	4.1 (1.8–9.5)	< 0.001
Lymph node status	pN+ /pNx vs. pN0	2.1 (1.3–3.5)	0.004	2.5 (1.4–4.2)	0.001
Adjuvant chemotherapy	yes vs. no	0.5 (0.3–0.8)	0.007	0.3 (0.2–0.5)	< 0.001
CD3+ T cells	continuous	0.3 (0.04–1.6)	0.20
CD4+ T cells	continuous	0.07 (0.0–1.6)	0.25
CD8+ T cells	continuous	0.4 (0.04–2.2)	0.38
Tregs	continuous	0.2 (0.03–0.8)	0.04	0.1 (0.0–0.7)	0.04
CD45+ immune cells	continuous	0.6 (0.2–1.6)	0.3
Fibroblasts	continuous	2.0 (0.6–5.9)	0.2
PD-L1+ immune cells	continuous	0.7 (0.17–2.4)	0.61
Proliferating tumor cells	continuous	0.6 (0.02–5.4)	0.76
Proliferating immune cells	continuous	0.2 (0.04–0.82)	0.04	0.2 (0.03–0.7)	0.02
Macrophages	continuous	4.0 (1.25–11.52)	0.01	10.9 (2.8–40.5)	< 0.001
K-means clusters for Tregs and macrophages	2 vs. 1	5.4 (1.54–18.8)	0.008	3.5 (1.0–12.5)	0.05
	2 vs. 3	2.1 (1.15–3.38)	0.02	2.4 (1.3–4.4)	0.006
	3 vs. 1	2.6 (0.8–8.3)	0.1	1.5 (0.4–4.8)	0.5

Variables with significant prediction on OS were added to the multivariate model adjusting for tumor, lymph node status and adjuvant chemotherapy. K-means clusters for macrophages and Tregs was performed using their ratio to all counted cells. Cluster 1: Treg high; Cluster 2: macrophage high; Cluster 3: Treg low and macrophage low

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