Expression and Clinical Significance of Th1/Th2/Th17 Cytokines and Lymphocyte Subsets in PCNSL

Figures & data

Table 1 The Base Information of Patients with PCNSL in This Study

Characteristics		Case (n)	Percentage (%)
Age/year
	<59	18	46.15
	≥59	21	53.85
Sex
	Male	14	35.9
	Female	25	64.1
LDH (U/L)
	<250	32	82.1
	≥250	7	17.95
Ki-67(%)
	<80%	8	20.51
	≥80%	31	79.49
EBV	+	10	25.64
C-Myc/Bcl-2	+	14	33.33
Treatment
	R-MDT	21	53.85
	R-BDT	13	35.9
	R-ZDT	3	7.69
	Symptomatic treatment	2	5.13
Clinical efficacy
	PR+CR	22	56.41
	PD+SD	17	43.59

Notes: “+” indicates positive. “PR+CR” indicates that the treatment was effective. “ PD+SD” indicates that the treatment was ineffective.

Table 2 Plasma Th1/Th2/Th17 Cytokine Levels [Pg/mL, M (Range)]

Group	n	IL-1β	IL-2	IL-4	IL-5	IL-6	IL-8
PCNSL	39	2.00(1.26~4.17)	1.33(0.98~1.82)	1.34(0.96~1.76)	1.12(0.91~1.52)	3.19(1.30~6.73)	2.30(1.41~3.91)
DLBCL	96	2.19(1.81~6.45)	1.23(0.96~2.09)	1.28(0.77~1.44)	1.25(0.98~2.00)	6.24(2.45~17.43)	2.30(1.70~4.38)
Healthy	13	0.41(0.00~2.56)	0.01(0.00~0.09)	0.25(0.23~0.31)	0.71(0.57~1.00)	0.82(0.58~1.15)	3.05(1.84~4.32)
H		13.332	34.597	34.529	13.732	28.653	0.498
P		0.001	<0.001	<0.001	0.001	<0.001	0.78
Group	n	IL-10	IL-12P	IL-17	TNF-α	IFN-α	IFN-γ
PCNSL	39	1.24(0.75~3.23)	1.35(1.09~1.77)	2.00(1.18~2.38)	1.67(1.19~2.29)	1.27(0.61~1.91)	3.00(1.87~5.38)
DLBCL	96	1.30(0.94~3.06)	1.19(0.84~1.77)	1.60(0.97~2.14)	2.21(1.03~2.35)	2.03(1.49~2.08)	3.93(2.39~8.63)
Healthy	13	0.44(0.37~0.49)	0.58(0.45~0.67)	0.38(0.31~0.54)	0.43(0.11~0.78)	0.47(0.10~1.21)	2.03(1.27~5.62)
H		28.432	22.021	30.782	26.322	18.943	3.781
P		<0.001	<0.001	<0.001	<0.001	<0.001	0.151

Notes: Kruskal–Wallis H-test, P<0.05 indicates that the difference is statistically significant.

Table 3 Plasma Th1/Th2/Th17 Cytokine Levels in 22 Patients with PCNSL in Effective Chemotherapy [Pg/mL, M (Range)]

Group	IL-1β	IL-2	IL-4	IL-5	IL-6	IL-8
Initial treatment	1.81(1.26~2.46)	1.09(0.80~1.33)	1.42(1.28~1.74)	1.11(1.08~1.51)	3.53(1.50~6.15)	2.30(1.58~3.53)
(PR+CR)	1.81(1.10~2.75)	1.59(1.43~2.76)	1.28(0.68~2.08)	1.08(0.79~1.94)	2.23(1.08~6.52)	2.30(0.90~4.10)
Z	−0.052	−2.902	−0.362	−0.24	−0.152	−0.414
P	0.959	0.004	0.717	0.811	0.879	0.679
Group	IL-10	IL-12P	IL-17	TNF-α	IFN-α	IFN-γ
Initial treatment	1.14(0.71~1.63)	1.75(1.14~1.77)	2.14(1.36~2.23)	2.21(1.23~2.26)	1.43(0.53~2.03)	3.00(1.77~4.00)
(PR+CR)	0.95(0.69~1.68)	1.23(1.09~1.70)	2.00(0.72~2.81)	1.40(1.02~2.93)	1.68(0.92~2.08)	3.29(2.36~6.11)
Z	−0.937	−0.362	−0.465	−0.284	−1.81	−1.065
P	0.349	0.717	0.642	0.776	0.07	0.287

Notes: Wilcoxon signed-rank, P<0.05 indicates that the difference is statistically significant. “PR+CR” indicates that the treatment was effective.

Table 4 Plasma Th1/Th2/Th17 Cytokine Levels in 17 Patients with PCNSL in Ineffective Chemotherapy [Pg/mL, M (Range)]

Group	IL-1β	IL-2	IL-4	IL-5	IL-6	IL-8
Initial treatment	2.33(1.18~4.94)	1.21(0.83~2.15)	1.21(0.82~1.82)	1.19(0.92~1.62)	3.06(1.07~14.86)	2.35(1.04~3.97)
(PD+SD)	2.46(0.92~5.52)	1.58(1.42~2.28)	1.19(0.70~1.83)	1.18(0.70~1.89)	1.35(0.51~5.85)	3.15(1.41~4.14)
Z	−0.401	−0.863	−0.141	−0.7	−1.54	−0.28
P	0.889	0.388	0.888	0.484	0.123	0.779
Group	IL-10	IL-12P	IL-17	TNF-α	IFN-α	IFN-γ
Initial treatment	1.63(0.83~4.01)	1.25(1.03~1.40)	1.56(1.12~2.24)	1.53(1.22~2.25)	1.16(0.62~1.69)	3.15(1.92~6.43)
(PD+SD)	1.11(0.92~1.26)	1.46(0.58~1.90)	1.46(0.58~1.90)	1.45(1.15~2.08)	1.46(0.58~1.90)	3.58(2.22~6.51)
Z	−0.28	−0.84	−0.28	−1.4	0	−0.42
P	0.779	0.401	0.779	0.161	1	0.674

Notes: Wilcoxon signed-rank, P<0.05 indicates that the difference is statistically significant. “PD+SD” indicates that the treatment was ineffective.

Figure 1 Distribution of IL-2 and IFN-α in patients with PCNSL and the control group.

Notes: (A) The level of IL-2 in PCNSL was significantly higher than that in the healthy people; (B) IFN-α levels in PCNSL were lower than those in DLBCL.

Figure 2 Comparison of IL-2 between initial and after treatment in patients with PCNSL.

Notes: (A) The level of IL-2 after effective treatment was significantly higher than before treatment; (B) there was no significant change in IL-2 levels before and after ineffective treatment.

Table 5 Levels of Lymphocyte Subsets at Initial Treatment [%, M (Range)]

Group	n	Treg	CD3^+	CD4^+	CD8^+	CD4+/CD8+	CD19^+
PCNSL	39	5.53(3.78~6.90)	74.93(69.33~79.09)	69.76(52.14~78.23)	19.40(13.91~27.35)	3.38(2.07~5.65)	11.47(4.16~16.92)
DLBCL	96	4.83(3.78~6.43)	74.82(65.96~81.93)	46.17(37.52~66.07)	24.30(20.61~35.53)	1.82(1.17~2.97)	7.21(3.77~11.02)
Healthy	27	2.56(2.27~2.91)	71.44(66.36~73.57)	37.53(32.86~44.76)	26.17(23.10~34.15)	1.50(0.95~1.91)	9.89(8.23~16.77)
H		39.168	4.132	32.961	14.245	30.819	9.837
P		<0.001	0.127	<0.001	<0.001	<0.001	0.007

Notes: Kruskal–Wallis H-test, P<0.05 indicates that the difference is statistically significant.

Table 6 Levels of Lymphocyte Subsets in 22 Patients with PCNSL in Effective Chemotherapy [%, M(Range)]

Group	Treg	CD3^+	CD4^+	CD8^+	CD4+/CD8+	CD19^+
Initial treatment	5.90(3.70~7.01)	75.67(71.94~79.22)	69.82(61.57~78.10)	20.82(14.29~25.67)	3.35(2.16~5.61)	11.43(8.49~15.62)
(CR+PR)	5.14(3.83~7.68)	83.12(77.33~90.05)	65.48(57.16~72.32)	27.26(22.62~35.02)	2.36(1.65~3.29)	0.04(0.02~0.11)
z	−0.876	−3.315	−1.673	−2.768	−2.403	−4.106
P	0.381	0.01	0.094	0.006	0.016	<0.001

Notes: Wilcoxon signed-rank, P<0.05 indicates that the difference is statistically significant. “PR+CR” indicates that the treatment was effective.

Table 7 Levels of Lymphocyte Subsets in 17 Patients with PCNSL in Ineffective Chemotherapy [%, M(Range)]

Group	Treg	CD3^+	CD4^+	CD8^+	CD4+/CD8+	CD19^+
Initial treatment	5.80(3.64~7.01)	72.78(64.22~77.09)	66.45(49.04~79.49)	16.09(12.75~29.29)	3.58(2.07~6.36)	12.22(3.97~20.59)
(PD+SD)	5.65(4.95~6.06)	85.29(77.68~93.88)	62.46(44.51~74.32)	23.41(18.51~40.71)	2.90(1.30~3.35)	0.01(0.00~0.03)
Z	−1.014	−2.756	−0.267	−2.134	−1.334	−2.803
P	0.31	0.006	0.155	0.033	0.033	0.005

Notes: Wilcoxon signed-rank, P<0.05 indicates that the difference is statistically significant. “PD+SD” indicates that the treatment was ineffective.

Figure 3 Distribution of Treg, CD3+, CD4+, CD8+, CD4/CD8, and CD19+ cells in patients with PCNSL and the control group.

Notes: (A) Treg levels were higher in PCNSL patients than in healthy subjects; (B) compared with DLBCL patients, PCNSL patients had higher CD19+ levels; (B), (D), (E) compared with DLBCL patients and healthy volunteers, PCNSL patients had higher levels of CD4+ and CD4+/CD8+ and lower levels of CD8+. ***P < 0.001.

Figure 4 Comparison of CD19+, CD3+, CD8+, CD4+/CD8+ between initial and after treatment in patients with PCNSL.

Notes: (A and B) After chemotherapy (whether effective or not), the level of CD19+ and CD4+/CD8+ decreased significantly compared with before chemotherapy; (C and D) after chemotherapy (whether effective or not), the level of CD8+ and CD3+ were significantly higher than that before chemotherapy.

Table 8 Analysis of Prognostic Factors

Factors		n	PFS M(95% CI) (Months)	Log Rank P	OS M(95% CI) (Months)	Log Rank P
Sex				0.802		0.886
	Male	14	18.00(14.43–46.28)		N
	Female	25	N		N
Age/year				0.71		0.123
	≥59	21	N		N
	<59	18	18.00(0.00–40.18)		N
IL-2(ng/L)				0.401		0.14
	≥1.33	14	3.00(0.00–9.78)		11.00(0.00–26.23)
	<1.33	15	N		N
CD19+ (%)				0.705		0.957
	≥11.47	20	18.00(0.00–41.50)		N
	<11.47	19	N		N
CD3+ (%)				0.405		0.773
	≥74.93	17	N		N
	<74.93	22	7.00(0.00–27.95)		N
CD8+ (%)				0.518		0.953
	≥19.40	18	N		N
	<19.40	21	18.00(0.00–47.43)		N
CD4+/CD8+				0.448		0.507
	≥3.38	20	5.00(-)		N
	<3.38	19	N		N
LDH(U/L)				0.351		0.957
	≥250	7	N		N
	<250	32	18.00(-)		N
Ki67(%)				0.141		0.984
	≥80	31	N		N
	<80	6	7.00(0.00–18.85)		N
C-Myc/Bcl-2	(+)	14	5.00(-)	0.532	N	0.922
EBV	(+)	10	18.00(4.04–31.96)	0.665	N	0.537

Notes: “+” indicates positive. “N” indicates positive median survival time was not reached.

Figure 5 Kaplan–Meier curves for progression-free and overall survival of all patients with PCNSL.

Notes: (A) progression-free survival curve; (B) overall survival curve.

Figure 6 Kaplan–Meier curves for progression-free and overall survival of all newly diagnosed patients with PCNSL.

Notes: (A and B) Kaplan–Meier curves for progression-free and overall survival of all newly diagnosed PCNSL patients with different level of IL-2; (C and D): Kaplan–Meier curves for progression-free and overall survival of PCNSL patients with different treatment options. Note (a) symptomatic treatment; (b) chemotherapy based on MTX(R-MDT); (c) chemotherapy based on osimertinib (R-BDT, R-BCT); (d) chemotherapy based on ibrutinib (R-ZDT).