Cytoplasmic and nuclear programmed death ligand 1 expression in peritumoral stromal cells in breast cancer: Prognostic and predictive value

Table 1 Baseline clinical and pathological information of patients with breast cancer

Breast cancer characteristics	Patients
	n	%
T stage
Т in situ	4	2.7
Т1в	11	7.5
Т1с	89	60.1
Т2	44	29.7
N stage
N0	81	54.7
N1	47	31.8
N2	16	10.8
N3	4	2.7
Histology
Ductal	134	90.5
Lobular	2	1.4
Papillary	4	2.7
Mucinous	4	2.7
Intraductal	4	2.7
Grade (G)
G1	16	10.8
G2	91	61.5
G3	41	27.7
Lymphovascular invasion
Yes	84	56.8
No	64	43.2
Perineural invasion
Yes	115	77.7
No	33	22.3
Molecular biological subtype
Luminal A	56	37.8
Luminal B HER2-negative	66	44.6
Luminal B HER2-positive	10	6.8
Non-luminal HER2-positive	10	6.8
TNBC	6	4.0
ER status
Negative	16	10.8
Positive	132	89.2
PR status
Negative	41	27.7
Positive	107	72.3
HER2 status
Negative	128	85.5
Positive	20	14.5

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; TNBC: Triple-negative breast cancer.

Table 2 Distribution of patients with programmed cell death protein 1 ligand 1-positive lymphocyte score < 3 and ≥ 3, n (%)

Breast cancer characteristics	PDCD1 LG1+ LF score		P value χ² tests
	< 3	≥ 3
T stage
Т in situ	0 (0)	4 (100)	0.50
Т1в	2 (18.2)	9 (81.8)
Т1с	27 (30.3)	62 (69.7)
Т2	12 (27.3)	32 (72.7)
N stage
N0	16 (19.7)	65 (80.3)	0.03
N1	19 (40.4)	28 (59.6)
N2	6 (37.5)	10 (62.5)
N3	0 (0)	4 (100)
Tumor grade (G)
G1	6 (37.5)	10 (62.5)	0.47
G2	26 (28.6)	65 (71.4)
G3	9 (21.9)	32 (78.1)
Lymphovascular invasion
No	18 (21.4)	66 (78.6)	0.05
Yes	23 (35.9)	41 (64.1)
Perineural invasion
No	31 (27.0)	84 (73.0)	0.70
Yes	10 (30.3)	23 (69.7)
ER status
Negative	3 (18.7)	13 (81.3)	0.39
Positive	38 (28.8)	94 (71.2)
PR status
Negative	15 (36.6)	2 (63.4)	0.13
Positive	26 (24.3)	81 (75.7)
HER2 status
HER2-	39 (30.5)	89 (69.5)	0.05
HER2+	2 (10.0)	18 (90.0)
Molecular biological subtype
Luminal A	18 (32.1)	38 (67.9)	0.26
Luminal B HER2-	20 (30.3)	46 (69.7)
Luminal B HER2+	0 (0)	10 (100)
Non-luminal HER2+	2 (20.0)	8 (80.0)
TNBC	1 (16.7)	5 (83.3)

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; TNBC: Triple negative breast cancer; PDCD1 LG1: Programmed cell death protein 1 ligand 1.

Table 3 Distribution of patients with and without nuclear expression of programmed cell death protein 1 ligand 1 in peritumoral lymphocytes, n (%)

Breast cancer characteristics	Nuclear expression of PDCD1 LG1 in peritumoral lymphocytes		P value χ² tests
	Absent	Present
T stage
Т in situ	0 (0)	4 (100)	0.69
Т1в	2 (18.2)	9 (8.18)
Т1с	12 (13.5)	77 (86.5)
Т2	4 (9.1)	40 (90.9)
N stage
N0	8 (9.9)	73 (90.1)	0.57
N1	8 (17.0)	39 (83.0)
N2	2 (12.5)	14 (87.5)
N3	0 (0)	4 (100)
Tumor grade (G)
G1	4 (25.0)	12 (75.0)	0.15
G2	8 (8.8)	83 (91.2)
G3	6 (14.6)	35 (85.4)
Lymphovascular invasion
No	6 (7.1)	78 (92.9)	0.03
Yes	12 (18.8)	52 (81.2)
Perineural invasion
No	12 (10.4)	103 (89.6)	0.23
Yes	6 (18.2)	27 (81.8)
ER status
Negative	0 (0)	16 (100)	0.11
Positive	18 (13.6)	114 (86.4)
PR status
Negative	10 (24.4)	31 (75.6)	0.004
Positive	8 (7.5)	99 (92.5)
HER2 status
HER2-	18 (14.1)	110 (85.9)	0.07
HER2+	0 (0)	20 (100)
Molecular biological subtype
Luminal A	6 (10.7)	50 (89.3)	0.20
Luminal B HER2-	12 (18.2)	54 (81.8)
Luminal B HER2+	0 (0)	10 (100)
Nonluminal HER2+	0 (0)	10 (100)
TNBC	0 (0)	6 (100)

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; PDCD1 LG1: Programmed cell death protein 1 ligand 1; TNBC: Triple-negative breast cancer.

Table 4 Distribution of patients with different degrees of programmed cell death protein 1 ligand 1-positive polymorphic cell infiltration, n (%)

Breast cancer characteristics	Degrees of PDCD1 LG1+ polymorphic cell infiltrations		P value χ² tests
	No, weak	Moderate, pronounced
T stage
Тis	2 (50.0)	2 (50.0)	0.02
Т1в	4 (36.4)	7 (63.6)
Т1с	59 (66.3)	30 (33.7)
Т2	36 (81.8)	8 (18.2)
N stage
N0	58 (71.6)	23 (28.4)	0.70
N1	31 (66.0)	16 (34.0)
N2	10 (62.5)	6 (37.5)
N3	2 (50.0)	2 (50.0)
Tumor grade (G)
G1	10 (62.5)	6 (37.5)	0.09
G2	68 (74.7)	23 (25.3)
G3	23 (56.1)	18 (43.9)
Lymphovascular invasion
No	60 (71.4)	24 (28.6)	0.34
Yes	41 (64.1)	23 (35.9)
Perineural invasion
No	79 (68.7)	36 (31.3)	0.82
Yes	22 (66.7)	11 (33.3)
ER status
Negative	5 (31.3)	11 (68.7)	0.0008
Positive	96 (72.7)	36 (27.3)
PR status
Negative	25 (60.9)	16 (39.0)	0.23
Positive	76 (71.0)	31 (29.0)
HER2 status
HER2 -	99 (77.3)	29 (22.7)	< 0.00001
HER2+	2 (10.0)	18 (90.0)
Molecular biological subtype
Luminal A	42 (75.0)	14 (25.0)	< 0.00001
Luminal B HER2-	54 (81.8)	12 (18.2)
Luminal B HER2+	0 (0)	10 (100)
Nonluminal HER2+	2 (20.0)	8 (80.0)
TNBC	3 (50.0)	3 (50.0)

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; TNBC: Triple-negative breast cancer; PDCD1 LG1: Programmed cell death protein 1 ligand 1.

Table 5 Distribution of patients with and without programmed cell death protein 1 ligand 1-positive fibroblastic stroma, n (%)

Breast cancer characteristics	PDCD1 LG1+ fibroblastic stroma		P value χ² tests
	Absent	Present
T stage
Т in situ	4 (100)	0 (0)	0.026
Т1в	6 (54.5)	5 (45.5)
Т1с	59 (66.3)	30 (33.7)
Т2	38 (86.4)	6 (13.6)
N stage
N0	60 (74.1)	21 (25.9)	0.73
N1	33 (70.2)	14 (29.8)
N2	12 (75.0)	4 (25.0)
N3	2 (50.0)	2 (50.0)
Tumor grade (G)
G1	10 (62.5)	6 (37.5)	0.61
G2	66 (72.5)	25 (27.5)
G3	31 (75.6)	10 (24.4)
Lymphovascular invasion
No	63 (75.0)	21 (25.0)	0.39
Yes	44 (68.8)	20 (31.2)
Perineural invasion
No	80 (69.6)	35 (30.4)	0.16
Yes	27 (81.8)	6 (18.2)
ER status
Negative	8 (50.0)	8 (50.0)	0.03
Positive	99 (75.0)	33 (25.0)
PR status
Negative	31 (75.6)	10 (24.4)	0.57
Positive	76 (71.0)	31 (29.0)
HER2 status
HER2 -	99 (77.3)	29 (22.6)	0.0005
HER2+	8 (40.0)	12 (60.0)
Molecular biological subtype
Luminal A	44 (78.6)	12 (21.4)	0.004
Luminal B HER2-	52 (78.8)	14 (21.2)
Luminal B HER2+	3 (30.0)	7 (70.0)
Non-luminal HER2+	5 (50.0)	5 (50.0)
TNBC	3 (50.0)	3 (50.0)

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; TNBC: Triple-negative breast cancer; PDCD1 LG1: Programmed cell death protein 1 ligand 1.

Table 6 Distribution of patients with and without programmed cell death protein 1 ligand 1 expression in the endothelial cells of tumor microvessels, n (%)

Breast cancer characteristics	PDCD1 LG1 expression in endothelial cells of tumor microvessels		P value χ² tests
	Absent	Present
T stage
Т in situ	2 (50.0)	5 (50.0)	0.06
Т1в	4 (36.4)	7 (63.6)
Т1с	57 (64.0)	32 (36.0)
Т2	2 (50.0)	2 (50.0)
N stage
N0	58 (71.6)	23 (28.4)	0.36
N1	27 (57.5)	20 (42.5)
N2	10 (62.5)	6 (37.5)
N3	2 (50.0)	2 (50.0)
Tumor grade
G1	12 (75.0)	4 (25.0)	0.41
G2	56 (61.5)	35 (38.5)
G3	29 (70.7)	12 (29.3)
Lymphovascular invasion
No	59 (70.2)	25 (29.8)	0.16
Yes	38 (59.4)	26 (40.6)
Perineural invasion
No	74 (64.3)	41 (35.7)	0.56
Yes	23 (69.7)	10 (30.3)
ER status
Negative	10 (62.5)	6 (37.5)	0.78
Positive	87 (65.9)	45 (34.1)
PR status
Negative	29 (70.7)	12 (29.3)	0.41
Positive	68 (63.5)	39 (36.5)
HER2 status
HER2 -	87 (68.0)	41 (32.0)	0.11
HER2+	10 (50.0)	10 (50.0)
Molecular biological subtype
Luminal A	38 (67.9)	18 (32.1)	0.13
Luminal B HER2-	46 (69.7)	20 (30.3)
Luminal B HER2+	3 (30.0)	7 (70.0)
Non-luminal HER2+	7 (70.0)	3 (30.0)
TNBC	3 (50.0)	3 (50.0)

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; PR: Progesterone receptor; TNBC: Triple-negative breast cancer.

Table 7 Results of the correlation analysis

Breast cancer characteristics	Gamma correlation	P value
Т stage	-0.240	0.022
Tumor grade	-0.160	0.133
Lymphovascular invasion	0.967	< 0.00001
Perineural invasion	0.380	0.003
ER status	0.606	0.00008
PR status	0.158	0.068
HER2 status	-0.359	0.029
Ki67 index	-0.114	0.147
Subtype	0.104	0.28
PDCD1 LG1+ LF score	-0.414	0.0004
Lymphocytes with nuclear expression of PDCD1 LG1	0.295	0.12
PDCD1 LG1+ polymorphic cellular infiltrations	0.151	0.105
PDCD1 LG1+ fibroblastic stroma	-0.029	0.796
PDCD1 LG1 expression in endothelial cells of tumor microvessels	0.289	0.011
SP142+ ICs	-0.283	0.033

ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; ICs: Immune cells; LFs: Lymphocytes; PDCD1 LG1: Programmed cell death protein 1 ligand 1; PR: Progesterone receptor; TNBC: Triple-negative breast cancer.

Table 8 Results of univariate and multivariate analyses

Breast cancer characteristics	Univariate analysis OR (95%CI)	P value	Multivariate analysis OR (95%CI)	P value
T stage
Tis-T1b	1	> 0.05
T1c	1.08 (0.43-5.81)
T2	0.54 (0.11-3.96)
Tumor grade (G)
G1	1	0.05
G2	1.78 (0.59-5.31)
G3	0.86 (0.26-2.87)
Lymphovascular invasion
No	1	< 0.00001	1	< 0.00001
Yes	60.26 (21.60-168.1)		82.73 (24.02-284.89)
Perineural invasion
No	1	0.047	1	> 0.05
Yes	2.23 (1.01-4.91)		0.28 (0.07-1.19)
ER status
Negative	1	0.03	1	> 0.05
Positive	4.08 (1.11-14.98)		7.35 (0.91-59.45)
PR status
Negative	1	> 0.05
Positive	1.08 (0.25-2.23)
HER2 status
HER2 -	1	> 0.05
HER2+	0.47 (0.17-1.30)
Molecular biological subtype
Luminal A	1	> 0.05
Luminal B HER2-	1.08 (0.53-2.21)
Luminal B HER2+	1.73 (0.44-6.81)
Нелюминальный HER2+	0.57 (0.16-1.21)
TNBC	1.15 (0.21-6.22)
Vessels with PDCD1 LG1 expression
absent	1	0.08
present	1.81 (0.91-3.59)
PDCD1 LG1+ LF score
< 3	1	0.018	1	> 0.05
≥ 3	0.41 (0.20-0.86)		0.46 (0.14-1.48)
Combination of LVI and PDCD1 LG1+ LF score
LVI- and PDCD1 LG1+ LF score < 3	1	0.004	1
LVI- and PDCD1 LG1+ LF score ≥ 3	0.13 (0.03-0.53)		0.11 (0.02-0.46)	0.003
LVI+ and PDCD1 LG1+ LF score < 3	9.50 (2.21-40.79)	0.002	11.09 (2.31-53.15)	0.003
LVI+ and PDCD1 LG1+ LF score ≥ 3	25.33 (5.48-117.1)	< 0.00001	44.35 (7.35-267.5)	< 0.00001

CI: Confidence interval; ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; LFs: Lymphocytes; LVI: Lymphovascular invasion; OR: Odds ratio; PDCD1 LG1: Programmed cell death protein 1 ligand 1; PR: Progesterone receptor; TNBC: Triple-negative breast cancer.

Table 9 Frequency of regional lymph nodes metastases according to the combinations of programmed cell death protein 1 ligand 1-positive lymphocytes score and lymphovascular invasion, n (%)

Combinations of PDCD1 LG1+ LF score and LVI	N stage		P value
	N0	N1-3
LVI- and PDCD1 LG1+ LF score < 3	12 (66.6)	6 (33.7)	0.00001
LVI- and PDCD1 LG1+ LF score ≥ 3	62 (93.9)	4 (6.1)
LVI+ and PDCD1 LG1+ LF score < 3	4 (17.4)	19 (82.6)
LVI+ and PDCD1 LG1+ LF score ≥ 3	3 (7.3)	38 (92.7)

LFs: Lymphocytes; LVI: Lymphovascular invasion; PDCD1 LG1: Programmed cell death protein 1 ligand 1.

Table 10 Research results assessing programmed death ligand 1 expression in breast cancer

Ref.	Number of patients/breast cancer subtype	Anti- programmed death-ligand 1 clone	Association of programmed death-ligand 1 expression with clinical and morphological characteristics of breast cancer	Association of programmed death-ligand 1expression with metastasis in regional lymph nodes
Li et al[36], 2018	112/Ductal BC/subtypes not specified	Polyclonal (Abcam)	Higher in G2-G3 (P = 0.03)	Associated with the presence of metastases
Cirqueira et al[40], 2021	6468/All subtypes	SP263, E1 L3N, 22C3, CAL 10, SP142, B7-H1, Polyclonal	Higher in high Ki67 (P = 0.005), HER2- BC (P = 0.001)	Associated with the absence of metastases
Zhou et al[41], 2018	136 Ductal BC/All subtypes	Monoclonal (ab213524)	Higher in luminal B HER2+, HER2+ non-luminal and TNBC (P = 0.001)	Not associated with the presence of metastases
Catacchio et al[42], 2019	180/All subtypes	Not specified	More often observed in lobular BC (P = 0.0287)	Not associated with the presence of metastases
Evangelou et al[43], 2020	45/All subtypes	Monoclonal E1 L3N	More often observed with a tumor size of more than 2 cm (P = 0.023), G3 (P = 0.017), ER- status (P = 0.001), PR- status (P = 0.002) and Ki67 more than 20% (P = 0.030)	Not associated with the presence of metastases
Zhang et al[37], 2020	4336/All subtypes	SP263, E1 L3N, 22C3, CAL 10, SP142, B7-H1, Polyclonal, NAT105, 28–8, ab58810, Abcam	More often observed in stage III (P = 0.015), ER- status (P = 0.000), HER2+ BC (P = 0.03), and higher levels of Ki67 (P = 0.000)	Associated with the presence of metastases
Kim et al[38], 2017	4578/All subtypes	Abcam	Higher in G3 (P = 0.001)	Associated with the presence of metastases
Zhang et al[39], 2017	2546, All subtypes	ab58810, NA, М1Н1	Higher in G3 (P = 0.001), ER- status (P = 0.008), TNBC (P = 0.001)	Associated with the presence of metastases

BC: Breast cancer; ER: Estrogen receptor; HER2: Human epidermal growth factor receptor 2; LVI: Lymphovascular invasion; PR: Progesterone receptor; TNBC: Triple-negative breast cancer.