Relationship of B7-H3 expression in tumor cells and tumor vasculature with FOXP3+ regulatory T cells in renal cell carcinoma

Figures & data

Figure 1 (A–C) Immunohistochemical staining for B7-H3 in tumor cells and the tumor vasculature of clear cell renal cell carcinoma. (A) Representative tumor with low B7-H3 expression in tumor cells and the tumor vasculature. (B) Representative tumor with high B7-H3 expression in tumor cells. (C) Representative tumor with high B7-H3 expression in the tumor vasculature. (D and E) Immunohistochemical staining for FOXP3 in tumor-infiltrating cells in clear cell renal cell carcinoma. (D) Representative tumor with a low FOXP3+ cell density. (E) Representative tumor with a high FOXP3+ cell density. Scale bar, 100 µm.

Table 1 Clinicopathological characteristics of clear cell renal cell carcinoma according to B7-H3 expression in tumor cells and the tumor vasculature

Variables		N of samples (%)	B7-H3 expression in tumor cells			B7-H3 expression in tumor vasculature
Low N=215 (85%)	High N=37 (15%)	P-values	Low N=115 (46%)	High N=137 (54%)	P-values
Age					0.92			0.029
<65 years		148 (59%)	126 (59%)	22 (59%)		76 (66%)	72 (53%)
≥65 years		104 (41%)	89 (41%)	15 (41%)		39 (34%)	65 (47%)
Sex					0.82			0.50
Male		187 (74%)	159 (74%)	28 (76%)		83 (72%)	104 (76%)
Female		65 (26%)	56 (26%)	9 (25%)		32 (28%)	33 (24%)
Tumor size					0.40			0.013
<40 mm		132 (52%)	115 (53%)	17 (46%)		70 (61%)	62 (45%)
≥40 mm		120 (48%)	100 (47%)	20 (54%)		45 (39%)	75 (55%)
Fuhrman grade					0.059			0.032
1–2		195 (77%)	171 (80%)	24 (65%)		96 (83%)	99 (72%)
3–4		57 (23%)	44 (20%)	13 (35%)		19 (17%)	38 (28%)
Pathological stage					0.45			0.015
I–II		221 (88%)	190 (88%)	31 (84%)		107 (93%)	114 (83%)
III–IV		31 (12%)	25 (12%)	6 (16%)		8 (7.0%)	23 (17%)
Performance status					0.65			0.76
0		234 (96%)	200 (96%)	34 (94%)		108 (96%)	126 (95%)
≥1		10 (4.1%)	8 (3.9%)	2 (5.6%)		4 (3.6%)	6 (4.6%)
FOXP3+ cell density					0.0069			0.0015
Low		126 (50%)	115 (53%)	11 (30%)		70 (61%)	56 (41%)
High		126 (50%)	100 (47%)	26 (70%)		45 (39%)	81 (59%)

Table 2 The associations of B7-H3 expression with clinicopathological features in clear cell renal cell carcinoma

		B7-H3 expression
		Univariable analysis		Multivariable analysis*
OR (95% CI)	P-values	OR (95% CI)	P-values
Factors to predict high B7-H3 expression in tumor cells
FOXP3+ cell density	high (vs low)	2.72 (1.31–5.99)	0.0069	2.93 (1.40–6.56)	0.0041
Fuhrman grade	3–4 (vs 1–2)	2.11 (0.97–4.42)	0.059	2.36 (1.07–5.09)	0.034
Tumor size (mm)	≥40 (vs <40)	1.35 (0.67–2.75)	0.40
Pathological stage	III–IV (vs I–II)	1.47 (0.51–3.68)	0.45
Sex	male (vs female)	1.10 (0.50–2.59)	0.82
Age (years)	<65 (vs ≥65)	1.04 (0.51–2.14)	0.92
Factors to predict high B7-H3 expression in tumor vasculature
FOXP3+ cell density	high (vs low)	2.25 (1.36–3.75)	0.0015	2.45 (1.46–4.17)	0.0007
Tumor size (mm)	≥40 (vs <40)	1.88 (1.14–3.13)	0.013	2.01 (1.19–3.41)	0.0084
Age (years)	≥65 (vs <65)	1.76 (1.06–2.95)	0.029	1.81 (1.07–3.10)	0.027
Pathological stage	III–IV (vs I–II)	2.70 (1.20–6.67)	0.015
Fuhrman grade	3–4 (vs 1–2)	1.94 (1.06–3.66)	0.032
Sex	male (vs female)	1.22 (0.69–2.14)	0.50

Notes: *The multivariable model initially included age (<65 years vs ≥65 years), sex (male vs female), tumor size (<40 mm vs ≥40 mm), Fuhrman grade (1–2 vs 3–4), pathological stage (I–II vs III–IV), and FOXP3+ cell density (low vs high). A backward stepwise elimination with a threshold P-value of 0.05 was used to select variables in the final model.

Abbreviations: CI, confidence interval; OR, odds ratio.

Table 3 B7-H3 expression and patient mortality* in clear cell renal cell carcinoma, stratified by density of tumor-infiltrating FOXP3+ cells

		Disease-specific mortality				Overall mortality
		Univariable analysis		Multivariable analysis**		Univariable analysis		Multivariable analysis**
HR (95% CI)	P-values	HR (95% CI)	P-values	HR (95% CI)	P-values	HR (95% CI)	P-values
All patients
B7-H3 expression in tumor cells: low (N=215)		1 (referent)		1 (referent)		1 (referent)		1 (referent)
B7-H3 expression in tumor cells: high (N=37)		2.37 (1.13–4.62)	0.024	1.67 (0.78–3.30)	0.18	1.84 (0.95–3.32)	0.068	1.44 (0.74–2.62)	0.28
FOXP3+ cell density: low
B7-H3 expression in tumor cells: low (N=115)		1 (referent)		1 (referent)		1 (referent)		1 (referent)
B7-H3 expression in tumor cells: high (N=11)		1.33 (0.21–4.72)	0.71	1.66 (0.25–6.70)	0.55	1.35 (0.32–3.90)	0.64	1.67 (0.38–5.14)	0.45
FOXP3+ cell density: high
B7-H3 expression in tumor cells: low (N=100)		1 (referent)		1 (referent)		1 (reference)		1 (reference)
B7-H3 expression in tumor cells: high (N=26)		2.98 (1.23–6.91)	0.017	1.51 (0.58–3.74)	0.39	2.06 (0.93–4.28)	0.074	1.30 (0.56–2.83)	0.53
All patients
B7-H3 expression in tumor vasculature: low (N=115)		1 (referent)		1 (referent)		1 (referent)		1 (referent)
B7-H3 expression in tumor vasculature: high (N=137)		2.48 (1.27–5.19)	0.0069	1.45 (0.73–3.12)	0.30	2.41 (1.38–4.42)	0.0017	1.86 (1.05–3.45)	0.035
FOXP3+ cell density: low
B7-H3 expression in tumor vasculature: low (N=70)		1 (referent)		1 (referent)		1 (referent)		1 (referent)
B7-H3 expression in tumor vasculature: high (N=56)		1.42 (0.54–3.79)	0.48	1.21 (0.45–3.28)	0.71	1.64 (0.74–3.71)	0.22	1.46 (0.65–3.31)	0.36
FOXP3+ cell density: high
B7-H3 expression in tumor vasculature: low (N=45)		1 (referent)		1 (referent)		1 (reference)		1 (reference)
B7-H3 expression in tumor vasculature: high (N=81)		4.86 (1.65–20.7)	0.0025	1.93 (0.57–8.90)	0.31	3.85 (1.61–11.4)	0.0016	3.13 (1.28–9.38)	0.010

Notes: *Cox proportional hazards regression models were used to calculate the HR and 95% CI. **The multivariable model initially included age (<65 years vs ≥65 years), sex (male vs female), tumor size (<40 mm vs ≥40 mm), Fuhrman grade (1–2 vs 3–4), pathological stage (I–II vs III–IV), and FOXP3+ cell density (low vs high). A backward stepwise elimination with a threshold P-value of 0.05 was used to select variables in the final model.

Abbreviations: CI, confidence interval; HR, hazard ratio.

Figure 2 Kaplan–Meier curves for disease-specific survival (A) and overall survival (B) in patients with clear cell renal cell carcinoma according to B7-H3 expression in tumor cells. Kaplan–Meier curves for disease-specific survival (C) and overall survival (D) in patients with clear cell renal cell carcinoma according to B7-H3 expression in the tumor vasculature.

Figure 3 Kaplan–Meier curves for disease-specific survival in patients with clear cell renal cell carcinoma according to B7-H3 expression in tumor cells (A and B) and the tumor vasculature (C and D), stratified by the density of tumor-infiltrating FOXP3+ cells. (A and C) Low FOXP3+ cell density group. (B and D) High FOXP3+ cell density group.