|Year : 2014 | Volume
| Issue : 3 | Page : 156-163
Interleukin-10 (−1082G/A) gene promotor polymorphism in Egyptian non-Hodgkin lymphoma patients: relation to other prognostic factors
Mohammed Ibrahim Sayed Ahmed MD , Doaa Ibrahim Hashad, Alshimaa E Hassen
Department of Clinical Pathology, Alexandria University, Alexandria, Egypt
|Date of Submission||13-Oct-2014|
|Date of Acceptance||15-Oct-2014|
|Date of Web Publication||31-Dec-2014|
Mohammed Ibrahim Sayed Ahmed
Clinical Pathology Department, Alexandria University, 22 Al-Geish Avenue, El-Shatby, Alexandria 21411, USA
Source of Support: None, Conflict of Interest: None
Background and Objectives Interleukin-10 (IL-10) is an important immunoregulatory cytokine in. It is part of a balanced network of cytokines and can be cancer promoting (immunosuppressive; stimulation of cell proliferation) or cancer inhibiting. The present study aimed at investigating possible association between IL10 (-1082G/A) gene promoter polymorphism in a sample of Egyptian Non-Hodgkin lymphoma patients and its relation to other prognostic factors.
Design and Methods A case control study was carried out on fifty NHL patients at first diagnosis and twenty age and sex- matched normal controls.
Results There was statistically insignificant correlation between IL10 (-1082G/A) genotype and age (P = 0.631), ESR (P = 0.087), serum LDH (P = 0.623), serum albumin (P = 0.108) serum β2 microglobulin (P = 0.578), and hemoglobin (P = 0.696) in patient group. The frequency of IL10-1082G allele was found to be higher in patients with NHL(28.3%) as compared with control subjects (12.5%)(P = 0.298) , with a higher frequency of IL-10-1082GG/GA genotypes in the former population (13.1%, 30.4%) versus (5.0%, 15.0%) χ2P = 0.073.
Conclusion The frequency of IL10-1082G allele was found to be higher in patients with NHL as compared with healthy controls, which is translated into a higher frequency of IL-10-1082GG/GA genotypes, thus may be associated with higher risk of developing NHL. There was no significant correlation between IL-10 polymorphism with other prognostic factors.
Keywords: IL10 (-1082G/A), non-Hodgkin lymphoma, prognosis
|How to cite this article:|
Sayed Ahmed MI, Hashad DI, Hassen AE. Interleukin-10 (−1082G/A) gene promotor polymorphism in Egyptian non-Hodgkin lymphoma patients: relation to other prognostic factors. Egypt J Haematol 2014;39:156-63
|How to cite this URL:|
Sayed Ahmed MI, Hashad DI, Hassen AE. Interleukin-10 (−1082G/A) gene promotor polymorphism in Egyptian non-Hodgkin lymphoma patients: relation to other prognostic factors. Egypt J Haematol [serial online] 2014 [cited 2020 Jan 28];39:156-63. Available from: http://www.ehj.eg.net/text.asp?2014/39/3/156/148251
| Introduction|| |
Interleukin (IL)-10 is an immunoregulatory cytokine produced mainly by the CD4 + Th2. However, it is also produced by some activated B cells, some Th1 cell-activated macrophages, and some nonhematopoietic sources (e.g. keratinocytes, colon carcinoma, and melanoma cells)  . IL-10 is an important immunoregulatory cytokine that inhibits T-cell function by suppressing the expression of proinflammatory cytokines such as TNFa, IL-1, IL-6, IL-8, and IL-12  . It also inhibits antigen-presenting cells by downregulating major histocompatibility complex class II and B7 expression , . In addition to these inhibitory actions, IL-10 promotes B-cell-mediated functions enhancing survival, proliferation, differentiation, and antibody production , . The human IL-10 gene is located on chromosome 1 and encodes for five exons (5.1 kb)  . The IL-10 promoter is highly polymorphic, with two informative microsatellites, IL-10.G and IL-10.R, 1.2 and 4 kb upstream of the transcription start site, and three frequent point mutations: _1082 (G/A), _819 (C/T), and _592 (C/A)  .
Tumor cells from B, T, and NK cell lymphoma can produce biologically active IL-10. As early as 1993, Blay et al.  investigated IL-10 serum levels using an ELISA, which detects both viral and human IL-10 in patients with active non-Hodgkin lymphoma (NHL) and healthy volunteers. They described the detection of IL-10 in serum from about 50% of these patients, but none of the control blood donors. IL-10 was detectable with a similar frequency in all subtypes of NHL and in all clinical stages, as well as in both EBV-seropositive and Epstein-Barr virus (EBV)-seronegative patients  .
In different lymphomas, increased IL-10 production has been reported and a negative prognostic factor for responsiveness toward treatment, as well as the disease-free and overall survival in patients with melanoma and solid tumors, particularly with lung, gastrointestinal, and renal-cell cancer  . Several groups reported on increased circulating IL-10 serum levels in gastric, colon, and renal-cell cancer patients. IL-10 serum levels commonly returned to normal in radically resected patients. Persistently elevated IL-10 serum levels after surgery predicted tumor recurrence. Moreover, a further significant increase in IL-10 serum levels has been observed in nonresponders after chemotherapy.
| Aim|| |
The present study aimed at investigating a possible association between the IL-10 (−1082G/A) gene promoter polymorphism in a sample of Egyptian NHL patients and its relation to other prognostic factors.
| Participants and methods|| |
The study was carried out on two groups: 50 NHL patients at first diagnosis and 20 age-matched and sex-matched normal controls recruited from the Clinical and Chemical Pathology Department of the Alexandria University Hospital. An informed consent was obtained from all the participants before the onset of the study.
Both groups were subjected to the following:
- Thorough assessment of history, with a special focus on the presence of B symptoms (night sweats, fever, and weight loss).
- Careful general examination, with a focus on lymphadenopathy (number of extranodal sites), organomegaly, and other system involvement (abdominal mass, neurological signs, gum hypertrophy, and skin lesions).
Blood samples were obtained from the antecubital vein with minimal stasis and using a complete aseptic technique for the following:
- Complete blood count  was performed on a five-part differential automated cell counter system KX-21N (Sysmex, Kobe, Japan).
- Liver function tests: serum alanine aminotransferase, aspartate aminotransferase, albumin, total protein, and total serum bilirubin ,,,, were assessed on an automated chemistry analyzer Dimension RXL (Dade Behring, Marburg, Germany').
- Renal function tests: blood urea and serum creatinine  .
- Erythrocyte sedimentation rate (ESR) was assayed using a Westergren tube  .
- Lactate dehydrogenase (LDH) level  .
- Serum β2-microglobulin assessment using the ELISA technique  .
- Detection of the IL-10 (−1082G/A) polymorphism  DNA extraction: genomic DNA was extracted from EDTA whole blood using the QIAamp DNA Blood Mini Extraction Kit (Qiagen, Hilden, Germany).
The procedure is designed to isolate genomic DNA from white blood cells. First, red blood cells are lysed by the addition of proteinase K to whole blood. This is followed by the addition of buffer AL, which acts to adjust the lysate buffering conditions to allow optimum binding of DNA onto the silica-gel membrane (mini spin column) during centrifugation and to ensure that proteins and other contaminants that can inhibit the PCR reaction are not retained on the silica-gel membrane. Lysis is completed after incubation for 10 min at 56°C, providing a maximum DNA yield. This is followed by the addition of ethanol, which works to dehydrate DNA, making it more amenable to binding onto the silica gel membrane. After application of the mixture to the QIAamp spin column, washing is carried out by buffer AW1 and then by buffer AW2 to remove residual contaminants. Using two types of washes improves the purity of the eluted DNA. The last step is DNA elution using buffer AE.
Preparation of the reagents
- 1.2 ml protease solvent was pipetted into a vial containing lyophilized Qiagen protease. This dissolved protease is stable for 2 months when stored at 2-8°C. As repeated freezing and thawing should be avoided, aliquots of protease were used.
- Buffer AW1 was prepared by adding 25 ml ethanol (96-100%), whereas buffer AW2 was prepared by adding 30 ml of ethanol (96-100%).
- Buffer AL was ready made and had to be shaken before use.
Procedure (protocol for purification of genomic DNA)
- 20 μl protease was pipetted into the bottom of a 1.5 ml microcentrifuge tube.
- 200 μl of the whole-blood sample (after it was brought to room temperature, completely thawed, and mixed thoroughly by gentle inversion) was added to the microcentrifuge tube.
- 200 μl buffer AL was added to the sample and then mixed by pulse vortex for 15 s for each sample.
- Incubation at 56°C was performed for 10 min.
- The microtube was then centrifuged briefly to remove drops from the inside of the lid.
- 6-200 μl ethanol (96-100%) was added to the sample and mixed thereafter by pulse vortex.
- The mixture was applied carefully to the QIAamp mini spin column (without wetting the rim) to which a collection tube was attached. The column was centrifuged at 8000 rpm for a minute. The tube containing the filtrate was discarded and the spin column was placed in another clean 2 ml collection tube.
- 500 μl of buffer AW1 was added without wetting the rim, and then the column was centrifuged at 8000 rpm for a minute. The tube containing the filtrate was discarded and the spin column was placed in another clean 2 ml collection tube.
- 500 μl of buffer AW2 was added and then the column was centrifuged at a speed of 14 000 rpm for 3 min.
- The collection tube was discarded and the spin column was placed in a sterile 1.5 ml microtube.
- 200 μl buffer AE was added, followed by incubation at room temperature for a minute, and then centrifugation was performed at 6000g (8000 rpm) for another minute.
- The eluted DNA was then stored at −20°C to be used for genotyping.
5′ Nuclease assay
Principle of the 5′ nuclease assay ,
This method utilizes the 5′-3′ exonuclease activity of Taq polymerase, where the enzyme cleaves 5′ terminal nucleotides of double-stranded DNA. A flourogenic oligonucleotide TaqMan probe is designed to anneal to a position between the two amplification primers in the PCR reaction. The probe is labeled with two florescent dyes: a reporter dye at the 5′ end and a quencher dye at the 3′ end. The quencher dye exerts a quenching effect on the fluorescence of the reporter dye when both dyes are in close proximity. During extension, Taq polymerase partially displaces the hybridized probe and then cleaves it at its 5′ end. Cleavage of the probe between the reporter and the quencher physically separates the two dyes, resulting in increased reporter fluorescence. This indicates that the probe-specific target has been amplified. For allelic discrimination, two probes with different reporters are used: one specific for the wild type and the other for the mutant allele. An increase in the fluorescent signal of a particular dye indicates homozygosity for the allele whose probe was labeled by that dye; an increase in both signals indicates heterozygosity.
- TaqMan Universal PCR Master Mix containing AmpliTaq Gold DNA polymerase enzyme. TaqMan Universal PCR Master Mix was 2× concentrated, containing, in addition to the polymerase enzyme, dNTPs and PCR buffer (with 3 mmol/l MgCl 2 ).
- Genotyping assay mix contained.
- Probe (FAM) specific for the IL-10 −1082G (CTTCCCCCTCCCAAA) allele (wild allele).
- Probe (VIC) specific for the IL-10 −1082A (CTTCCCCTTCCCAAAG) allele (mutant allele).
- Forward primer (5′-CAAATCCAAGACAAC ACTACTAAGGC-3′).
- Reverse primer (5′-GGGTGGAAGAAGTT GAAATAACAAG-3′).
- DNA samples, probes, and primers were removed from storage temperature (−20°C) and thawed at room temperature.
- DNA samples were assessed for DNA concentration and quality using a NanoDrop 2000 Spectrophotometer (Thermo Scientific, Waltham, MA, USA) (ng/μl).
- Specific fluorescent dye-labeled (FAM and VIC) minor groove binder (MGB) probes were diluted to a final concentration of 200 nmol/l.
- PCR primers were used in the reaction, with a final concentration of 600 nmol/l.
- Two tubes for each sample (one for each probe) were prepared.
The first tube contained:
The second tube contained:
Note that a single tube technique was used in IL-10 (−1082G/A) genotyping for the detection of both alleles in the same tube, but yielding no appropriate results; thus, two tubes were used to detect a single allele per tube.
Polymerase chain reaction thermal cycling program on Rotor Gene-Q (QIAGEN GmbH, Hilden, German)
- Initial hold (one cycle): 95°C for 10 min.
- Multistep cycling (45 cycles):
- Denaturing: 95°C for 15 s.
- Annealing/extension: 60°C for 60 s.
The fluorescence profile of each well was assayed on a Rotor Gene-Q detection system. Sequence detection software determines the contribution of each dye to the final fluorescence signal and displays the results on a scatter plot of the wild allele versus the mutant allele and thus the genotype can be determined. A marked increase was observed in only in VIC dye fluorescence: homozygosity for the A allele; only in FAM dye fluorescence: homozygosity for the G allele; both fluorescence signals: heterozygosity.
The data were collected and entered into the personal computer. Statistical analysis was carried out using the statistical package for social sciences (SPSS, version 20; SPSS Inc., Chicago, Illinois, USA) software. To calculate the arithmetic mean and SD for categorized parameters, the χ2 -test was used, whereas for numerical data, a t-test was used to compare two groups and for more than two groups, the ANOVA test was used. The level of significance was 0.05.
| Results|| |
[Table 1] shows the comparison between the two groups studied according to the demographic data. The control group included 10 (50%) men and 10 (50%) women, mean age 50.40 ± 13.3 years. The NHL patients included 27 (54%) men and 23 (46%) women, mean age 53.11 ± 10.13 years. There were statistically insignificant differences between the two groups in sex and age (P = 0.336, 0.401, respectively).
|Table 1 Comparison between the two studied groups according to the demographic data|
Click here to view
Erythrocyte sedimentation rate levels
The mean level of ESR in NHL patients was 67.74 ± 44.14 mm/h (range 22.0-210.0 mm/h). It was higher than that in 20 controls (14.90 ± 4.01 mm/h, range 8.0-20.0 mm/h). There was a statistically significant difference in ESR in patients compared with the control group (P = 0.0025).
Lactate dehydrogenase level
The mean level of LDH in NHL patients was 549.48 ± 213.55 U/l (range 254.0-945.0 U/l). It was higher than in the control group (291.0 ± 45.87 U/l, range 234.0-401.0 U/l). There was a statistically significant difference in LDH in patients compared with the control group (P = 0.0036).
The mean level of β2-microglobulin in NHL patients was 6.39 ± 2.951 μg/ml (range 2.0-11.40 μg/ml). It was higher than that in 20 controls (1.09 ± 0.61 μg/ml, range 0.32-2.60 μg/ml). There was a statistically significant difference in β2-microglobulin levels in NHL patients compared with the control group (P = 0.001). Estimation of serum albumin levels was performed in both NHL patients and the control group.
The mean serum albumin level in NHL patients was 2.77 ± 0.35 g/l (range 2.50-4.0 g/l) compared with a mean level of 4.20 ± 0.65 g/l (range 3.40-5.30 g/l) in the control group. There was a statistically significant difference in serum albumin levels in patients compared with the control group (P ≤ 0.001).
The mean hemoglobin (Hb) level in NHL patients was 9.61 ± 1.50 g/dl (range 6.5-13.0 g/dl) compared with a mean level of 12.83 ± 0.59 (range 12.0-14.0 g/dl) in the control group. There was a statistically significant difference in the Hb level in patients compared with the control group (P 0≤ 0.001).
IL-10 (−1082G/A) polymorphism
The allele G frequency was higher in NHL patients (27%) than in the control group (12.5%), whereas the allele A frequency was higher in the control group (87.5%) than in the group of NHL patients (73%). G/G and G/A genotypes was over-represented (16, 30%) in patients with NHL compared with the control group (5, 15%), whereas genotype A/A was over-represented in the control group (80%) than in the NHL patients (54%). There were insignificant differences between cases and controls in allele frequencies (P = 0.108) and genotype distribution (0.301) ([Table 2]).
|Table 2 Comparison between the two studied groups according to the interleukin-10 (−1082G/A) genotype and allele frequencies|
Click here to view
Sex, age, β2-microglobulin, albumin, hemoglobin, erythrocyte sedimentation rate, and lactate dehydrogenase) was insignificant (P > 0.05) and significant with ESR (P = 0.013) ([Table 3]).
|Table 3 Relation between the interleukin 10 (−1082G/A) genotype with demographic data in the patient group|
Click here to view
| Discussion|| |
Lymphoma is a generic term used to describe malignant expansion of any of the lymphoid cell series  . NHLs are clonal lymph proliferative disorders that are heterogonous with respect to their clinical presentation, pathogenesis, and biologic behavior  . Although the exact cause of NHL remains unknown, emerging evidence suggests that NHL is a complex disorder caused by the interaction of the immune system, multiple genes, and environmental factors  .
IL-10 is an important immunoregulatory cytokine in humans. IL-10 is part of a balanced network of cytokines and can be cancer promoting (immunosuppressive, stimulation of cell proliferation) or cancer inhibiting. IL-10 is produced by several cells including normal and neoplastic B cells, stimulated monocytes/macrophages, and subsets of T cells. IL-10 has been implicated in certain infectious diseases, autoimmunity, transplantation tolerance, and tumorigenesis  .
Variable associations between IL-10 production capacity and either the IL-10 microsatellite alleles, single-nucleotide polymorphisms (SNP), or SNP haplotypes in the 7-kb IL-10 5′-flanking region have been reported. Several studies have reported that proximal IL-10 promoter polymorphisms may be related to an increased risk of a diverse range of diseases. This indicates that genetic variations within the IL-10 gene locus are relevant in vivo , .
Recent reports have provided evidence that a risk of developing NHL or the clinical outcome of patients with diffuse large B-cell lymphoma (DLBCL) might be related to certain IL-10 promoter gene variations. An epidemiologic multicenter study described the IL-10 −3538A regulatory SNP to be associated with an increased risk of developing NHL. In one study, it was reported that proximal genotypes or haplotypes with a low IL-10 expression are a risk factor for aggressive lymphoma, whereas a second study suggested that genotypes with high expression potential are a risk factor for developing lymphoma in patients with AIDS , .
The aim of this study was to investigate a possible association between the IL-10 (−1082G/A) promoter polymorphism in NHL lymphoma patients compared with healthy controls and its relation to other prognostic variables (age, sex, ESR, serum LDH, serum albumin, albumin, and serum β2-microglobulin).
The present study included seventy participants divided into two groups: the first group included 50 NHL patients and the second group included 20 age-matched and sex-matched healthy controls. The control group included 50% men and 50% women, mean age 45.4 ± 13.29 years. The NHL patients included 56.5% men and 43.5% women, mean age 54.9 ± 9.38 years. There were statistically insignificant differences between the two groups in sex and age (P = 0.669, 0.294, respectively).
In the present study, there was a statistically significant decrease in serum albumin levels in patients compared with the control group (P ≤ 0.001). The low serum albumin level in NHL patients can be attributed to cytokine release in the circulation. IL-6 inhibits the hepatic synthesis of albumin, causing hypoalbuminemia  . Talaat et al.  reported that low serum albumin in NHL patients at diagnosis is associated with a low response rate and a high death rate during treatment.
The laboratory evaluations of patients showed significant abnormalities in the levels of LDH and β2-microglobulin associated with hypoalbuminemia compared with the control group (P < 0.001). This finding is in agreement with several previous large-scale studies that reported that hypoalbuminemia, elevation of serum β2-microglobulinand, and elevated LDH level were important prognostic factors for lymphoma ,,, .
Increases in LDH may be used both in the clinical diagnosis of cancer patients and in monitoring tumor size following chemotherapy. β2-Microglobulin and LDH were found at increased concentrations and were considered strong markers of disease activity  .
β2-Microglobulin levels tended to increase with advanced NHL. Recently, many investigators have concluded that β2-microglobulin levels alone or in combination with serum LDH levels were critical and independent factors in predicting prognosis  .
Increased serum LDH is a common finding in patients with cancer and is generally attributed to tumor aggressiveness or a high tumor burden. High total serum LDH carries a poor prognosis in myeloma, childhood acute lymphoblastic leukemia (ALL), melanoma, lung adenocarcinoma, and colorectal carcinoma. Many authors have shown that high serum LDH is a major prognostic factor in patients with NHL  .
Total serum LDH is one of the parameters of the International Prognostic Index used in patients with NHL  . LDH is a useful marker of tumor activity and can be measured serially to assess response to treatment. Most high-risk lymphomas have high levels of LDH initially; a rapid decrease reflects tumor responsiveness and is associated with a more favorable prognosis  .
In the present study, ESR values were significantly elevated in patients than in the control group (P < 0.001). This finding is in agreement with that of Elahi et al.  who reported that ESR more than 50 mm/h was associated with poor survival in patients with aggressive NHL. Elevated ESR in NHL patients could be explained by the fact that IL-6 is released in the circulation and acts as an inducer of hepatic fibrinogen synthesis, the major determinant of ESR  .
The Hb concentration in NHL patients ranged between 6.50 and 13.0 g/dl, with a mean level of 9.61 ± 1.50 g/dl, compared with a mean level of 11.45 ± 1.21 (range 9.50-13.50 g/dl) in the control group. There were statistically significant decreases in Hb level in patients compared with the control group (P ≤ 0.001). According to the WHO criteria, anemia is diagnosed as Hb concentration less than 13 g/dl for an adult male and 12 g/dl for an adult female. Accordingly, mild anemia was one of the features in our NHL patients  .
There are many mechanisms of anemia in NHL; it may be anemia of chronic disease, autoimmune hemolytic anemia, hypersplenism, bone marrow (BM) infiltration by lymphoma cells, or because of bone marrow suppression after chemotherapy  .
Conlan et al.  reported that newly diagnosed NHL patients with an Hb concentration less than 12 g/dl have increased death rates during treatment; this is also related to a shortened relapse-free interval and overall survival. The short survival of anemic patients has been found to be independent of bone marrow infiltration by lymphoma cells.
In the present study, no correlation was found between the IL-10 −1082G/A promoter polymorphism and other prognostic variables (age P = 0.631, ESR P = 0.087, serum LDH P = 0.623, serum β2-microglobulin P = 0.578, and Hb P = 0.736). The frequency of the IL-10 −1082G allele was found to be higher in patients with NHL (28.3%) compared with healthy control participants (12.5%) (P = 0.298), which translated into a higher frequency of IL-10 −1082GG/GA genotypes in the former population (13.1, 30.4% vs. 5.0, 15.0%) (χ2 ; P = 0.073). These findings are in agreement with previous reports ,, , thus showing that the GG genotype may be associated with a higher risk of developing NHL.
Lech-Maranda et al.  investigated whether the IL-10 polymorphism influences this cytokine production as well as well as the incidence and outcome of DLBCL. A moderate excess of the IL-10 −1082 allele was found among the patients with DLBCL compared with ethnically matched healthy control participants. These data suggest that the presence of the IL-10 −1082 allele may contribute toward the genetic background of DLBCL occurrence.
Purdue et al.  studied whether IL-10 variants 3757A and 1082G were associated with an increased risk of DLBCL. Additional haplotype analysis of these polymorphisms showed that the increased risk of DLBCL was restricted to the AG haplotype (containing both risk alleles) rather than the TG haplotype (containing only the 1082 risk allele), suggesting that 3575A is more important than 1082G in influencing the risk of lymphoma.
Talaat et al.  studied the impact of the IL-10 (1082G/A, rs1800896 and 819 C/T, rs1800871) gene promoter polymorphism on the susceptibility of Egyptians to develop DLBCL, the major type of NHL. Genotyping polymorphism was performed using a sequence-specific primer PCR in 100 Egyptian DLBCL patients and 119 normal controls. An insignificant change in IL-10 (−1082 and 819) genotypes was recorded. Although the A allele was slightly decreased in DLBCL patients, it did not reach statistical significance. The GT haplotype was significantly elevated in NHL patients (P<0.05).
Cunningham et al.  studied whether polymorphisms in the IL-10 gene polymorphism play a role in predisposing an individual to lymphoma. They analyzed the frequencies of three single base substitutions in the IL-10 promotor in patients with aggressive lymphoma (DLBCL n = 46, other aggressive histologies n = 17), Hodgkin's disease (n = 44), or low/intermediate-grade lymphoma (n = 46), compared with healthy controls.
The frequency of the low IL-10-producing AA allele at position (−1082) was significantly higher in patients with aggressive lymphoma compared with the controls (odds ratio 1.974, 95% confidence interval 1.066-3.655, P = 0.0344). No association was found between IL-10 genotypes and Hodgkin's lymphoma or less aggressive forms of lymphoma. Thus, the polymorphism in the IL-10 gene promoter that is associated with a low IL-10-producing phenotype may influence susceptibility to aggressive forms of lymphoma or may contribute toward the pathogenesis of this disease. The variation documented in our work from that of Cunningham et al.  work could be attributed to the ethnic differences and to the limited number of our patients.
| Conclusion|| |
The IL-10 (−1082G/A) polymorphism may be relatively common among Egyptian NHL patients. No significant difference was detected between NHL patients and healthy controls in the IL-10 (−1082G/A) genotype distribution. The frequency of the IL-10 −1082G allele was found to be higher in patients with NHL compared with healthy controls, which translated into a higher frequency of IL-10 −1082GG/GA genotypes in the former population, and may thus be associated with higher risk of developing NHL. There was no significant correlation between the IL-10 polymorphism with other prognostic factors, age, ESR, serum LDH, serum albumin serum β2-microglobulin, and Hb levels.
The variation documented in this work from other studies in different populations could be attributed to the limited number of participants and ethnic differences. Therefore, larger prospective studies are needed to confirm our findings. Other IL-10 gene polymorphisms, including distal loci -7400InDel, -6752AT (rs6676671), and -6208CG (rs10494879) in comparison with proximal loci −3538AT (rs1800890), −1087AG (rs1800896), and -597AC (rs1800872), should be studied.
| Acknowledgements|| |
Conflicts of interest
There are no conflicts of interest.
| References|| |
Del Prete G, De Carli M, Almerigogna F, Giudizi MG, Biagiotti R, Romagnani S. Human IL-10 is produced by both type 1 helper (Th1) and type 2 helper (Th2) T cell clones and inhibits their antigen-specific proliferation and cytokine production. J Immunol
De Waal Malefyt R, Haanen J, Spits H, Roncarolo MG, te Velde A, Figdor C, et al
. Interleukin 10 (IL-10) and viral IL-10 strongly reduce antigen-specific human T cell proliferation by diminishing the antigen-presenting capacity of monocytes via downregulation of class II major histocompatibility complex expression. J Exp Med 1991
Ding L, Linsley PS, Huang LY, Germain RN, Shevach EM. IL-10 inhibits macrophage costimulatory activity by selectively inhibiting the up-regulation of B7 expression. J Immunol
Rousset F, Garcia E, Defrance T, Péronne C, Vezzio N, Hsu DH, et al
. Interleukin 10 is a potent growth and differentiation factor for activated human B lymphocytes. Proc Natl Acad Sci USA
Jones KD, Aoki Y, Chang Y, Moore PS, Yarchoan R, Tosato G. Involvement of interleukin-10 (IL-10) and viral IL-6 in the spontaneous growth of Kaposi's sarcoma herpesvirus-associated infected primary effusion lymphoma cells. Blood
Eskdale J, Kube D, Tesch H, Gallagher G. Mapping of the human IL10 gene and further characterization of the 5¢ flanking sequence. Immunogenetics
Platzer C, Volk HD, Platzer M. 5¢ Noncoding sequence of human IL-10 gene obtained by oligo-cassette PCR walking. DNA Seq
Blay JY, Burdin N, Rousset F, Lenoir G, Biron P, Philip T, et al
. Serum interleukin-10 in non-Hodgkin's lymphoma: a prognostic factor. Blood
Dummer R, Heald PW, Nestle FO, Ludwig E, Laine E, Hemmi S, Burg G. Sézary syndrome T-cell clones display T-helper 2 cytokines and express the accessory factor-1 (interferon-gamma receptor beta-chain). Blood
Bain B, Bate I. Basic haematological techniques. In: Lewis SM, Bain BJ, editors. Dacie and Lewis practical haematology
. 9th ed. Churchill Livingstone Elsevier: Harcourt Publishing Limited
; 2001. 19-46.
Nuttall K, Klee G. Analytes of hemoglobin metabolism - porphyrins, iron, and bilirubin. In: Burtis CA, Ashwood ER, editors. Tietz fundamental of clinical chemistry
. 5th ed. WB Saunders Company
; 2001. II. 6016.
Henderson A, Enymes I, Burtis C, Ashwood E. Tietz fundamental of clinical chemistry
. 5th ed. WB Saunders Company
; 2001; I
Ahb W. Tietz clinical guide to laboratory tests
. 4th ed. St. Louis: E.B Saunders Company; 2006.
Dufour DR, Lott JA, Nolte FS, Gretch DR, Koff RS, Seeff LB. Diagnosis and monitoring of hepatic injury. I. Performance characteristics of laboratory tests. Clin Chem
Turner DM, Williams DM, Sankaran D, Lazarus M, Sinnott PJ, Hutchinson IV. An investigation of polymorphism in the interleukin-10 gene promoter. Eur J Immunogenet
Hui L, DelMonte T, Ranade K. Genotyping using the TaqMan assay. Curr Protoc Hum Genet 2008; Chapter 2
Ranade K, Chang MS, Ting CT, Pei D, Hsiao CF, Olivier M, et al
. High-throughput genotyping with single nucleotide polymorphisms. Genome Res 2001; 11
Green AR, Hoffbrand AV, Catovsky D, Tuddenham EG. Postgraduate haematology
. 6th ed. Wiley-Blackwell; 2011.
Cerhan JR, Liu-Mares W, Fredericksen ZS, Novak AJ, Cunningham JM, Kay NE, et al
. Genetic variation in tumor necrosis factor and the nuclear factor-kappaB canonical pathway and risk of non-Hodgkin's lymphoma. Cancer Epidemiol Biomarkers Prev
Lichtman MA, Beutler E, Kaushansky K, Kipps TJ, Seligsohn U, Prchal J. Williams hematology
. 8th ed. McGraw-Hill Medical. McGraw-Hill Professional; 2010. 1846-1863.
Abbas AK, Lichtman AH, Pillai S, Baker A. Properties and overview of immune response. In: Abbas AK, Lichtman AH, Pillai S, editors. Cellular and molecular immunology
. 7th ed. Philadelphia: Elsevier Saunders; 2012. 2-329.
Wang J, Ding Q, Shi Y, Cao Q, Qin C, Zhu J, et al
. The interleukin-10 −1082 promoter polymorphism and cancer risk: a meta-analysis. Mutagenesis 2012; 27
Wong HL, Breen EC, Pfeiffer RM, Aissani B, Martinson JJ, Margolick JB, et al
. Cytokine signaling pathway polymorphisms and AIDS-related non-Hodgkin lymphoma risk in the multicenter AIDS cohort study. AIDS 2010; 24
Talaat RM, Abdel-Aziz AM, El-Maadawy EA, Abdel-Bary N. Interleukin 10 gene promoter polymorphism and risk of diffuse large B cell lymphoma (DLBCL). Egypt J Med Hum Genet
Yildirim R, Gundogdu M, Erdem F, Kiki I, Bilici M. The levels of serum C-reactive protein, beta 2 microglobulin, ferritin, lactate dehydrogenase and some specific proteins in patients with non-Hodgkin's lymphoma before and after treatment. Eurasian J Med
Ferraris AM, Giuntini P, Gaetani GF. Serum lactic dehydrogenase as a prognostic tool for non-Hodgkin lymphomas. Blood
Navarro JT, Ribera JM, Oriol A, Vaquero M, Romeu J, Batlle M, et al
. International prognostic index is the best prognostic factor for survival in patients with AIDS-related non-Hodgkin's lymphoma treated with CHOP. A multivariate study of 46 patients. Haematologica 1998; 83
Chen W, Luo RC, Fan WW, Ma SD. Clinical value of combined detection of LDH, TPS, CEA and beta2-MG in patients with non-Hodgkin's lymphoma. Nan Fang Yi Ke Da Xue Xue Bao
Elahi MM, McMillan DC, McArdle CS, Angerson WJ, Soukop M, Johnstone J, Sattar N The systemic inflammatory response predicts overall and cancer specific survival in patients with malignant lymphoma. Med Sci Monit 2005; 11
Conlan MG, Armitage JO, Bast M, Weisenburger DD. Clinical significance of hematologic parameters in non-Hodgkin's lymphoma at diagnosis. Cancer 1991; 67
[ Lech-Maranda E, Baseggio L, Bienvenu J, Charlot C, Berger F, Rigal D, et al
. Interleukin-10 gene promoter polymorphisms influence the clinical outcome of diffuse large B-cell lymphoma. Blood
Purdue, MP, Lan Q, Kricker A, Grulich AE, Vajdic CM, Turner J, et al
. Polymorphisms in immune function genes and risk of non-Hodgkin lymphoma: findings from the New South Wales non-Hodgkin Lymphoma Study. Carcinogenesis
Cunningham LM, Chapman C, Dunstan R, Bell MC, Joske DJ. Polymorphisms in the interleukin 10 gene promoter are associated with susceptibility to aggressive non-Hodgkin's lymphoma. Leuk Lymphoma
[Table 1], [Table 2], [Table 3]