Epstein-Barr virus copy number in peripheral blood mononuclear cells predicts prognosis in diffuse large B cell lymphoma

Figures & data

Table 1. Baseline characteristics of patients according to EBV DNA copy number.

	All patients (n = 84)	EBV-DNA (copies/ml)
		<500 (n = 46)	500–10^4 (n = 27)	≥10^4 (n = 11)	p-trend
Total case, no. (%)	84 (100)	46 (54.76)	27 (32.14)	11 (13.10)
Median age, years (range)	53 (46–62)	51 (37–63)	57 (51–62)	53 (48–67)	.049
Older than 60 years	25 (29.8)	13 (28.3)	8 (29.6)	4 (36.4)
60 years or less	59 (70.2)	33 (71.7)	19 (70.4)	7 (63.6)	.636
Gender, male (%)	39 (46.4)	15 (32.6)	16 (59.3)	8 (72.7)	.003
B symptoms presence, no. (%)	39 (46.4)	23 (50)	10 (37.0)	6 (54.5)	.669
ECOG performance status, no. (%)
0–1	67 (79.8)	37 (80.4)	22 (81.5)	8 (72.7)
2–4	17 (20.2)	9 (19.6)	5 (18.5)	3 (27.3)	.748
Extranodal involvement, no. (%)
0–1	44 (55.0)	25 (56.8)	17 (68.0)	2 (18.2)
≥2	36 (45.0)	19 (43.2)	8 (32.0)	9 (81.8)	.280
LDH, no. (%)
Upper limit of normal or below	27 (32.9)	12 (26.7)	12 (46.2)	3 (27.3)
Over the upper limit of normal	55 (67.1)	33 (73.3)	14 (53.8)	8 (72.7)	.332
Bone marrow infiltration, no. (%)	16 (20.3)	10 (23.3)	4 (16.0)	2 (18.2)	.517
Ann Arbor stage, no. (%)
I–II	21 (25.6)	11 (24.4)	7 (26.9)	3 (27.3)
III–IV	61 (74.4)	34 (75.6)	19 (73.1)	8 (72.7)	.796
IPI score, no. (%)
0–2	38 (48.7)	18 (41.9)	16 (64.0)	4 (40.0)
3–5	40 (51.3)	25 (58.1)	9 (36.0)	6 (60.0)	.345
Histologic subtype, no. (%)
Germinal center B cell-like	29 (43.3)	19 (48.7)	8 (40.0)	2 (25.0)
Non-germinal center B cell-like	38 (56.7)	20 (51.3)	12 (60.0)	6 (75.0)	.236

ECOG: Eastern Cooperative Oncology group; IPI: international prognostic index; LDH: lactate dehydrogenase.

Table 2. Pathologic characteristics of patients according to EBV DNA copy number.

	All patients^a	EBV-DNA (copies/ml)^b
		<500	500–10^4	p^c	≥10^4	p^c
CD20	69/70 (99)	38 (97.4)	24 (100)	1.000	7/0	1.000
CD79a	46/47 (97.9)	25 (96.2)	16 (100)	1.000	5 (100)	1.000
BCL-2	58/67 (86.6)	31 (83.8)	20 (87.0)	1.000	7 (100)	.568
BCL-6	62/72 (86.1)	34 (85.0)	20 (83.3)	1.000	8 (100)	.558
CD10	22/68 (32.4)	16 (42.1)	6 (26.1)	.207	0 (0)	.087
Mum1	44/62 (71.0)	24 (68.6)	15 (71.4)	.822	5 (83.3)	.804
CD5	16/67 (23.9)	12 (30.8)	1 (4.8)	.045	3 (42.9)	.849
CD30	10/45 (22.2)	4 (16.7)	5 (29.4)	.556	1 (25)	1.000
EBER	5/46 (10.9)	2 (8.3)	2 (11.1)	1.000	1 (25)	.382

a Presented as no. of positive/all (percentage).

b Presented as no. of positive (percentage).

c Compared with the group with EBV-DNA <5 × 10² copies/ml.

EBV: Epstein-Barr virus, EBER: EBV-encoded small RNA.

Table 3. Tumor cytogenetic features of patients according to EBV DNA copy number.

	All patients^a	EBV-DNA (copies/ml)^b
		<500	500–10^4	p^c	≥10^4	p^c
P53 deletion	6/43 (14.0)	3 (15.8)	1 (7.1)	.620	2 (20)	1.000
Bcl-6 rearrangement	14/45 (31.1)	6 (28.6)	6 (42.9)	.477	2 (20)	1.000
c-myc rearrangement	2/44 (4.5)	0 (0)	0 (0)	1.000	2 (22.2)	.083
bcl-2 rearrangement	3/45 (6.7)	2 (9.1)	0 (0)	.519	1 (10)	1.000
Polyploid	8/46 (17.4)	3 (13.6)	3 (21.4)	.658	2 (20)	.637
c-myc copy number variation	5/44 (11.4)	4 (18.2)	1 (7.7)	.630	0 (0)	.295
bcl-2 copy number variation	3/46 (6.5)	2 (9.1)	1 (7.1)	1.000	0 (0)	1.000

a Presented as no. of positive/all (percentage).

b Presented as no. of positive (percentage).

c Compared with the group with EBV-DNA <5 × 10² copies/ml.

EBV: Epstein-Barr virus.

Figure 1. Frequencies and distribution of gene mutations in patients with DLBCL. A total of 41 DLBCL cases were performed with next-generation sequencing. Twenty-seven patients (65.85%) presented at least one variant. The bar chart above presented mutation in each patient. The bar chart nearby presented mutation frequency of each gene.

Figure 2. Kaplan–Meier’s estimates of (A) progression-free survival and (B) overall survival for the negative (<500 copies/ml), low (500–10⁴ copies/ml), and high EBV-CN group (≥10⁴ copies/ml), (C) progression-free survival, and (D) overall survival for positive group (EBV-DNA ≥ 500 copies/ml) and negative group (EBV-DNA <500 copies/ml), (E) progression-free survival and (F) overall survival for high EBV-CN group (EBV-DNA ≥ 1 × 10⁴ copies/ml) and not significantly increased EBV-CN group (EBV-DNA <1 × 10⁴ copies/ml).

Table 4. Univariate and multivariate analyses of prognostic factors.

	Univariate analysis			Multivariate analysis
Prognostic factor	HR^b	95.0%CI^c	p value	HR	95.0%CI	p value
Age
≥60	2.078	0.835–5.170	.116	1.064	0.340–3.325	.915
<60	1			1
Extranodal involvement
≥2	2.652	1.005–6.996	.049	1.354	0.389–4.714	.634
0–1	1			1
Serum LDH
>ULN^a	5.023	1.152–21.893	.032	2.536	0.544–11.817	.236
<ULN	1			1
Ann Arbor stage
III–IV	7.053	0.940–52.911	.057	6.006	0.611–58.989	.124
I–II	1			1
ECOG
2–4	3.374	1.323–8.605	.011	2.797	1.009–7.749	.048
0–1	1			1
Chemotherapy
Yes	0.085	0.029–0.249	.000	0.099	0.020–0.478	.004
No	1			1
EBV DNA, copies/ml
≥1 × 10^4	5.007	1.949–12.861	.001	5.841	1.660–20.548	.006
<1 × 10^4	1			1

a Upper limit of normal.

b Hazard ratio.

c Confidence interval.

LDH: lactate dehydrogenase, ECOG: Eastern Cooperative Oncology group, EBV: Epstein-Barr virus, HR: hazard ratio, CI: confidence interval.