J Hematol

Journal of Hematology, ISSN 1927-1212 print, 1927-1220 online, Open Access

Article copyright, the authors; Journal compilation copyright, J Hematol and Elmer Press Inc

Journal website http://www.thejh.org

Original Article

Volume 1, Number 4-5, October 2012, pages 77-88

Oxidative Stress in Patients With Early Stage Chronic Lymphocytic Leukemia, Assessment and Correlation With Prognostic Factors

Figure 1. Comparison of OS biomarkers values of early stage CLL and healthy control groups. Results are presented as mean ± standard deviation (erythrocytes biomarkers GSH, GSSG, GSSG/GSH, CAT, GPx, SOD and plasma biomarkers GSSG, GSSG/GSH) or geometric mean ± antilog standard deviation (erythrocytes biomarkers TBARS, GR and plasma biomarkers GSH, TBARS) in the healthy control group (n = 37) and early stages of CLL patients group (n = 37). *Statistical differences between healthy control and early stages of CLL patients with a P value < 0.001, ^†with a P value = 0.001, ^‡with a P value = 0.013, ^§with a P value = 0.016.

Figure 2. Frequency distribution of the SOS. Healthy control individuals present a normal distribution centered on zero. The frequency distribution of the SOS on early stage CLL did not follow a normal distribution, since the score was displaced towards positive values.

Figure 3. Correlation of OS biomarkers with evaluated prognostic of early stage CLL. (A) No significant differences could be observed in the levels of OS biomarkers CD38 (A), ZAP70 (B) and cytogenetic karyotipe (C) in relation to the prognostic. Regarding β-2M (D), a tendency of higher SOS could be observed in the group of patients with an adverse prognosis (β-2M > 2.4 mg/L) without significance.

**Table 1.** Scoring Criteria for Each Biomarker to Derive the Score of Oxidative Stress (SOS)
Biomarkers	Value > than upper limit	Value < than lower limit
SOS: -1, antioxidant status; +1, oxidative stress; 0, antioxidant-prooxidant equilibrium.
Antioxidant Enzymes
SOD	-1	+1
CAT	-1	+1
GR	-1	+1
GPx	-1	+1
GSH, GSSH, GSSG/GSH
GSH erythrocytes	-1	+1
GSSG erythrocytes	+1	-1
GSSG/GSH erythrocytes	+1	-1
Lipid peroxidation products
TBARS erythrocytes	+1	-1
TBARS plasma	+1	-1

Table 1. Scoring Criteria for Each Biomarker to Derive the Score of Oxidative Stress (SOS)

Biomarkers

Value > than upper limit

Value < than lower limit

SOS: -1, antioxidant status; +1, oxidative stress; 0, antioxidant-prooxidant equilibrium.

Antioxidant Enzymes

SOD

-1

CAT

-1

GPx

-1

GSH, GSSH, GSSG/GSH

GSH erythrocytes

-1

GSSG erythrocytes

-1

GSSG/GSH erythrocytes

-1

Lipid peroxidation products

TBARS erythrocytes

-1

TBARS plasma

-1

**Table 2.** Characteristics of Known Prognostic Factors of the 37 CLL Patients Included in the Study
*combination of positivity for CD5 and CD23, low positivity for Igs and CD22, and negativity for FMC7.
Prognostic factor (favorable vs. adverse)	Patients with favorable prognostic	Patients with adverse prognostic
LDT > 12 months v. ≤ 12 months	37/37 (100%)	0/37 (0%)
Lymphocyte morphology, Typical v atypical	30/37 (81.08%)	7/37 (18.92%)
LDH ≤ 500 UI/L v. > 500 UI/L	31/37 (83.78%)	6/37 (16.22%)
β-2M ≤ 2.4 mg/L v. > 2.4 mg/L	24/34 (70.59%)	10/34 (29.41%)
Immunophenotypic Score* 5 v. ≤ 4	29/34 (85.29%)	5/34 (14.71%)
CD38 Negative v. positive	25/32 (78.13%)	7/32 (21.82%)
ZAP-70 Negative v. positive	26/27 (96.30%)	1/27 (3.70%)
Cytogenetics 13, +12, normal cariotip v. 11, 17	28/30 (100%)	2/30 (100%)
lymphadenophatic areas by CT ≤ 2 v. > 2	36/37 (97.30%)	1/37 (2.701%)
Liver or spleen enlargement by CT scan Absence v. presence	36/37 (97.30%)	1/37 (2.70%)

Table 2. Characteristics of Known Prognostic Factors of the 37 CLL Patients Included in the Study

*combination of positivity for CD5 and CD23, low positivity for Igs and CD22, and negativity for FMC7.

Prognostic factor (favorable vs. adverse)

Patients with favorable prognostic

Patients with adverse prognostic

LDT > 12 months v. ≤ 12 months

37/37 (100%)

0/37 (0%)

Lymphocyte morphology, Typical v atypical

30/37 (81.08%)

7/37 (18.92%)

LDH ≤ 500 UI/L v. > 500 UI/L

31/37 (83.78%)

6/37 (16.22%)

β-2M ≤ 2.4 mg/L v. > 2.4 mg/L

24/34 (70.59%)

10/34 (29.41%)

Immunophenotypic Score* 5 v. ≤ 4

29/34 (85.29%)

5/34 (14.71%)

CD38 Negative v. positive

25/32 (78.13%)

7/32 (21.82%)

ZAP-70 Negative v. positive

26/27 (96.30%)

1/27 (3.70%)

Cytogenetics 13, +12, normal cariotip v. 11, 17

28/30 (100%)

2/30 (100%)

lymphadenophatic areas by CT ≤ 2 v. > 2

36/37 (97.30%)

1/37 (2.701%)

Liver or spleen enlargement by CT scan Absence v. presence

36/37 (97.30%)

1/37 (2.70%)

**Table 3.** Values of OS Biomarkers in Function of the 3 Groups of NAPF
Biomarkers	Without NAPF (n = 8)	1-2 NAPF (n = 22)	3-4 NAPF (n = 8)
Values are expressed as means ± SD (n) for the OS biomarkers of oxidative stress.
Erythrocytes
GSH (mmol/g Hb)	1.85 ± 1.08	1.69 ± 1.12	1.48 ± 0.68
GSSG (mmol/g Hb)	1.78 ± 1.15	1.75 ± 0.80	1.96 ± 1
GSSG/GSH	1.21 ± 0.82	1.25 ± 0.92	1.81 ± 1.59
TBARS (nmol/g Hb)	3.44 ± 3.34	3.54 ± 3.51	4.02 ± 3.58
CAT (mmol/min/g Hb)	175.5 ± 50.22	176.7 ± 43.28	143.15 ± 78.11
GPx (mmol/min/g Hb)	21.39 ± 4.56	19.46 ± 5.02	16.21 ± 8.58
GR (mmol/min/g Hb)	2.91 ± 2.48	3.22 ± 2.73	2.14 ± 1.17
SOD (U/g Hb)	1,052.6 ± 496.3	1,141.31 ± 483.22	702.74 ± 404.17
Plasma
GSH (mmol/mL)	15.01 ± 5.57	22.2 ± 16.88	24.86 ± 15.89
GSSG (mmol/mL)	29.15 ± 14.55	32.6 ± 16.23	32.63 ± 15.56
GSSG/GSH	2.04 ± 0.94	2.12 ± 1.96	1.66 ± 1.12
TBARS (nmol/mL)	3.16 ± 2.4	3.20 ± 2.49	3.81 ± 2.88
SOS	3.00 ± 1.41	2.81 ± 0.93	3.38 ± 1.41
ORAC	24.36 ± 5.19	32.35 ± 16.37	37.88 ± 23.59

Table 3. Values of OS Biomarkers in Function of the 3 Groups of NAPF

Biomarkers

Without NAPF (n = 8)

1-2 NAPF (n = 22)

3-4 NAPF (n = 8)

Values are expressed as means ± SD (n) for the OS biomarkers of oxidative stress.

Erythrocytes

GSH (mmol/g Hb)

1.85 ± 1.08

1.69 ± 1.12

1.48 ± 0.68

GSSG (mmol/g Hb)

1.78 ± 1.15

1.75 ± 0.80

1.96 ± 1

GSSG/GSH

1.21 ± 0.82

1.25 ± 0.92

1.81 ± 1.59

TBARS (nmol/g Hb)

3.44 ± 3.34

3.54 ± 3.51

4.02 ± 3.58

CAT (mmol/min/g Hb)

175.5 ± 50.22

176.7 ± 43.28

143.15 ± 78.11

GPx (mmol/min/g Hb)

21.39 ± 4.56

19.46 ± 5.02

16.21 ± 8.58

GR (mmol/min/g Hb)

2.91 ± 2.48

3.22 ± 2.73

2.14 ± 1.17

SOD (U/g Hb)

1,052.6 ± 496.3

1,141.31 ± 483.22

702.74 ± 404.17

Plasma

GSH (mmol/mL)

15.01 ± 5.57

22.2 ± 16.88

24.86 ± 15.89

GSSG (mmol/mL)

29.15 ± 14.55

32.6 ± 16.23

32.63 ± 15.56

GSSG/GSH

2.04 ± 0.94

2.12 ± 1.96

1.66 ± 1.12

TBARS (nmol/mL)

3.16 ± 2.4

3.20 ± 2.49

3.81 ± 2.88

SOS

3.00 ± 1.41

2.81 ± 0.93

3.38 ± 1.41

ORAC

24.36 ± 5.19

32.35 ± 16.37

37.88 ± 23.59