TLR4 expression correlated with PD-L1 expression indicates a poor prognosis in patients with peripheral T-cell lymphomas

Figures & data

Figure 1 Gene expression profiling analysis in PTCL. (A) Hierarchical clustering of gene expression profiles generated from five AITL samples and five normal T-cell samples. DEGs were selected with a fold change >2 and P-value < 0.001 among the mRNA expression profiling; (B) Ten samples are clustered according to the expression of 17 differentially expressed genes; (C and D) Top 30 enrichment of GO terms and KEGG pathways for differentially expressed genes; (E) The PPI network of 17 DEGs was constructed. Red cycle nodes represent genes, and the edges represent interaction between genes, the width of edges represents the combined score.

Table 1 Correlation between TLR4 and PD-L1 expression and various clinicopathological features

Characteristics	Cases, N (%)	TLR4		P-value	PD-L1		P-value
Low	High		Low	High
Age (years)
<60	89 (61.8)	53 (59.6)	36 (40.4)	0.706	50 (56.1)	39 (43.8)	0.537
≥60	55 (38.2)	31 (56.4)	24 (43.6)		28 (50.9)	27 (49.1)
Gender
Male	93 (64.6)	52 (55.9)	41 (44.1)	0.426	55 (59.1)	38 (40.9)	0.744
Female	51 (35.4)	32 (62.7)	19 (37.3)		23 (29.5)	28 (42.4)
Histological type
PTCL-NOS	17 (11.8)	7 (41.2)	10 (58.8)	0.203	7 (41.2)	10 (58.8)	0.148
AITL	88 (61.1)	53(60.2)	35 (39.8)		44 (50.0)	44 (50.0)
ALK^+ ALCL	20 (13.9)	10 (50.0)	10 (50.0)		14 (70.0)	6 (30.0)
ALK^− ALCL	19 (13.2)	14 (73.7)	5 (26.3)		13 (68.4)	6(31.6)
Primary site
Nodal	116 (80.5)	74 (63.8)	42 (36.2)	0.007	65 (56.1)	51 (43.9)	0.360
Extranodal	28 (19.5)	10 (35.7)	18 (64.3)		13 (46.4)	15 (53.6)
Tumor size
<7.5 cm	133 (92.3)	76 (57.1)	57 (42.9)	0.314	69 (51.9)	64 (48.1)	0.055
≥7.5 cm	11 (7.7)	8 (72.7)	3 (27.3)		9 (81.8)	2 (18.2)
No. of extranodal involvement(s)
0–1	15 (10.4)	11 (73.3)	4 (26.7)	0.213	12 (80.0)	3 (20.0)	0.034
>1	129 (89.6)	73 (56.6)	56 (43.4)		66 (51.2)	63 (48.8)
Ann Arbor stage
I–II	39 (27.1)	28 (71.8)	11 (28.2)	0.039	23 (58.9)	16 (41.0)	0.480
III–IV	105 (72.9)	56 (53.3)	49 (46.7)		55 (52.4)	50 (47.6)
B symptoms
Absent	63 (43.7)	32 (38.1)	31 (51.7)	0.106	29 (46.1)	34 (53.9)	0.084
Present	81 (56.3)	52 (61.9)	29 (48.3)		49 (60.5)	32 (39.5)
ECOG score
0–1	127 (88.2)	73 (57.5)	54 (42.5)	0.266	68 (53.5)	59 (46.5)	0.682
2–4	17 (11.8)	11 (64.7)	6 (35.3)		10 (58.2)	7 (41.2)
IPI score
Lower	96 (66.7)	56 (58.3)	40 (41.7)	1.000	51 (53.1)	45 (46.9)	0.723
Higher	48 (33.3)	28 (58.3)	20 (41.7)		27 (56.2)	21 (43.8)
LDH levels (U/L)
<250 U	75 (52.1)	42 (56.0)	33 (44.0)	0.637	41 (17.1)	34 (82.9)	0.900
≥250	69 (47.9)	42 (60.9)	27 (39.1)		37 (53.6)	32 (46.4)
HGB levels (g/L)
<120	42 (29.2)	21 (50.0)	21 (50.0)	0.005	19 (45.2)	23 (54.8)	0.168
≥120	102 (70.8)	63 (61.8)	39 (38.2)		59 (57.5)	43 (42.5)
PLT count (10^9/L)
<100	9 (6.3)	5 (55.6)	4 (44.4)	0.654	8 (88.9)	1 (11.1)	0.031
≥100	134 (93.7)	79 (58.2)	56 (41.8)		70 (51.5)	65 (48.5)
Leukocyte count (10^9/L)
<7.20	68 (47.2)	37 (54.4)	31 (45.6)	0.367	36 (52.9)	32 (47.1)	0.780
≥7.20	76 (52.8)	47 (61.8)	29 (38.2)		42 (55.3)	34 (44.7)
Ki-67 (%)
<60	84 (58.3)	48 (57.1)	36 (42.9)	0.732	50 (59.5)	34 (40.5)	0.127
≥60	60 (41.7)	36 (60.0)	24 (40.0)		28 (46.7)	32 (53.3)

Abbreviations: AITL, angioimmunoblastic T-cell lymphoma; ALCL, anaplastic large cell lymphoma; IPI, international prognostic index; LDH, lactate dehydrogenase; PD-L1, programmed cell death-ligand 1; PTCL-NOS, peripheral T-cell lymphomas not otherwise specified; TLR4, toll-like receptor 4.

Figure 2 Representative images of TLR4 and PD-L1 immunohistochemical staining of PTCL tissues. (A) Negative and (B) positive TLR4 expression in PTCL tissues; (C) Negative and (D) positive PD-L1 expression in PTCL tissues.

Table 2 Correlation between TLR4 expression and PD-L1 expression

TLR4 expression	No. of cases	PD-L1 expression	P-value
Low expression High expression
Low expression	84	63 21	<0.001
High expression	60	15 45

Abbreviations: PD-L1, programmed cell death-ligand 1; TLR4, toll-like receptor 4.

Figure 3 Kaplan Meier analysis for PFS and OS based on TLR4 and PD-L1 expression in PTCL patients. (A) Kaplan-Meier analysis for PFS based on TLR4 expression in patients with PTCL (log-rank test, P=0.021); (B) Kaplan-Meier analysis for OS based on TLR4 expression in patients with PTCL (log-rank test, P=0.016); (C) Kaplan-Meier analysis for PFS based on PD-L1 expression in patients with PTCL (log-rank test, P=0.002); (D) Kaplan-Meier analysis for OS based on PD-L1 expression in patients with PTCL (log-rank test, P<0.001).

Figure 4 Kaplan Meier analysis for PFS and OS based on TLR4 and PD-L1 expression in AITL patients. (A) Kaplan-Meier analysis for PFS based on TLR4 expression in patients with AITL (log-rank test, P=0.208); (B) Kaplan-Meier analysis for OS based on TLR4 expression in patients with AITL (log-rank test, P=0.027); (C) Kaplan-Meier analysis for PFS based on PD-L1 expression in patients with AITL (log-rank test, P=0.091); (D) Kaplan-Meier analysis for OS based on PD-L1 expression in patients with AITL (log-rank test, P=0.021).

Table 3 Prognostic factors affecting the progression free survival of patients with PTCLs

Characteristics		HR (95% CI) for Progression Free Survival
		Univariate HR				Multivariate HR for TLR4				Multivariate HR for PD-L1
HR	Lower	Upper	P-value	HR	Lower	Upper	P-value	HR	Lower	Upper	P-value
TLR4	High vs. Low	1.560	1.032	2.357	0.035	1.780	1.167	2.716	0.007
PD-L1	High vs. Low	1.639	1.071	2.510	0.023					1.718	1.121	2.633	0.013
Age (years)	>60 vs. ≤60	1.416	0.928	2.161	0.106
Gender	Female vs. Male	0.662	0.423	1.036	0.071
Primary site	Extranodal vs. Nodal	0.831	0.484	1.427	0.503
Tumor size	≥7.5 cm vs. <7.5 cm	0.633	0.256	1.565	0.322
No. of extranodal involvement(s)	0–1 vs. >1	0.781	0.377	1.618	0.506
Ann Arbor stage	I–II vs. III–IV	0.501	0.305	0.824	0.006
B symptoms	Present vs. Absent	1.637	1.059	2.525	0.026	1.582	1.105	2.466	0.043	1.584	1.017	2.466	0.042
ECOG score	2–4 vs. 0–1	1.574	0.832	2.979	0.163
IPI score	Higher vs. Lower	1.429	0.923	2.213	0.110
LDH levels (U/L)	≥250 vs. <250	1.315	0.870	1.989	0.194
HGB levels (g/L)	<120 vs. ≥120	0.768	0.475	1.240	0.280
PLT count (10^9/L)	<100 vs. ≥100	1.576	0.633	3.924	0.329
Leukocyte count (10^9/L)	<7.20 vs. ≥7.20	0.638	0.419	0.972	0.036	0.673	0.434	1.043	0.076	0.684	0.446	1.051	0.083
Ki-67 (%)	≥60 vs. <60	1.012	0.665	1.539	0.955

Abbreviations: CI, confidence interval; HR; IPI, international prognostic index; LDH, lactate dehydrogenase; PD-L1, programmed cell death-ligand 1; PTCLs, peripheral T-cell lymphomas; TLR4, toll-like receptor 4.

Table 4 Prognostic factors affecting the overall survival of patients with PTCLs

Characteristics		HR (95% CI) for Progression Free Survival
		Univariate HR				Multivariate HR for TLR4				Multivariate HR for PD-L1
HR	Lower	Upper	P-value	HR	Lower	Upper	P-value	HR	Lower	Upper	P-value
TLR4	High vs. Low	1.672	1.043	2.678	0.033	1.686	1.017	2.797	0.043
PD-L1	High vs. Low	2.379	1.425	3.971	0.001					2.591	1.545	4.345	<0.0001
Age (years)	>60 vs. ≤60	1.696	1.050	2.740	0.031
Gender	Female vs. Male	0.538	0.311	0.929	0.026	0.561	0.316	0.997	0.049	0.446	0.256	0.777	0.004
Primary site	Extranodal vs. Nodal	0.657	0.345	1.251	0.201
Tumor size	≥7.5 cm vs. <7.5 cm	0.687	0.249	1.892	0.467
No. of extranodal involvement(s)	0–1 vs. >1	0.707	0.284	1.764	0.458
Ann Arbor stage	I–II vs. III–IV	0.491	0.280	0.863	0.013
B symptoms	Present vs. Absent	1.421	0.873	2.312	0.157
ECOG score	2–4 vs. 0–1	2.419	1.182	4.947	0.016
IPI score	Higher vs. Lower	1.918	1.174	3.134	0.009	1.934	1.163	3.217	0.011	1.963	1.183	3.256	0.009
LDH levels (U/L)	≥250 vs. <250	1.116	0.693	1.796	0.651
HGB levels (g/L)	<120 vs. ≥120	0.963	0.564	1.645	0.891
PLT count (10^9/L)	<100 vs. ≥100	1.360	0.424	4.363	0.605
Leukocyte count (10^9/L)	<7.20 vs. ≥7.20	0.567	0.350	0.919	0.021	0.631	0.379	1.049	0.076	0.576	0.353	0.940	0.027
Ki-67 (%)	≥60 vs. <60	0.660	0.403	1.079	0.097

Abbreviations: CI, confidence interval; HR; IPI, international prognostic index; LDH, lactate dehydrogenase; PD-L1, programmed cell death-ligand 1; PTCLs, peripheral T-cell lymphomas; TLR4, toll-like receptor 4.