|Year : 2016 | Volume
| Issue : 6 | Page : 1123-1128
|Evaluation of interleukin-2, interleukin-8, and tumor necrosis factor-like weak inducer of apoptosis in hemodialysis and renal transplant patients and healthy controls
Nasar Alwahaibi1, Halima Alissaei1, Amal Al-Kalbani1, Nadia Alabri2, Zainab Allawati2, Mohammed Albalooshi3
1 Department of Allied Health Sciences, College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Sultanate of Oman
2 Department of Pathology, College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Sultanate of Oman
3 Department of Microbiology and Immunology, College of Medicine and Health Sciences, Sultan Qaboos University, Muscat, Sultanate of Oman
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|Date of Web Publication||28-Nov-2016|
| Abstract|| |
There is an increasing evidence which suggests that cytokines have an important role in hemodialysis (HD) and renal transplant patients. The aim of this study was to evaluate the levels of interleukin (IL)-2, IL-8 and tumor necrosis factor-like weak inducer of apoptosis (TWEAK) in HD and renal transplant patients and healthy controls. A total of 152 individuals participated in this study over two years including 53 HD patients, 45 renal transplant patients, and 54 healthy controls. Serum levels of IL-2, IL-8 and TWEAK were measured by enzyme-linked immunosorbent assay. The mean ages for HD, renal transplant patients, and healthy controls were 51 ± 13.6, 41.1 ± 13.0 and 40.73 ± 7.0 years, respectively. Serum levels of IL-2, IL-8 and TWEAK were significantly higher in HD patients compared to healthy controls. IL-2 concentrations were significantly higher in renal transplant patients compared to healthy controls. In conclusion, the findings of this study showed high levels of IL-2, IL-8 and TWEAK in HD patients and normal levels of IL-8 and TWEAK but high levels of IL-2 in renal transplant patients.
|How to cite this article:|
Alwahaibi N, Alissaei H, Al-Kalbani A, Alabri N, Allawati Z, Albalooshi M. Evaluation of interleukin-2, interleukin-8, and tumor necrosis factor-like weak inducer of apoptosis in hemodialysis and renal transplant patients and healthy controls. Saudi J Kidney Dis Transpl 2016;27:1123-8
|How to cite this URL:|
Alwahaibi N, Alissaei H, Al-Kalbani A, Alabri N, Allawati Z, Albalooshi M. Evaluation of interleukin-2, interleukin-8, and tumor necrosis factor-like weak inducer of apoptosis in hemodialysis and renal transplant patients and healthy controls. Saudi J Kidney Dis Transpl [serial online] 2016 [cited 2021 Dec 6];27:1123-8. Available from: https://www.sjkdt.org/text.asp?2016/27/6/1123/194594
| Introduction|| |
There is an increasing evidence suggesting that cytokines have an important role in chronic kidney disease (CKD). Despite progress in hemodialysis (HD) and renal transplantation, these patients are still at a high risk of cardiovascular disease.  Cardiovascular disease is a major cause of morbidity and mortality in HD and renal transplant patients. 
Patients with CKD are subject to continuous inflammatory triggers either from the dialysis procedure or from other factors such as diabetes mellitus, hypertension, and hyperlipidemia. There is an activation of immune system in CKD with many mediators such as complements, procoagulants, fibrinolytics, and cytokines that initiate inflammation in CKD.  Many studies suggest a link between cytokines and disease severity. , Cytokines are proteins secreted mainly by leukocytes and other body cells in response to antigenic cause. Interleukin-2 (IL)-2 and IL-8 have been proposed as proinflammatory cytokines in CKD, but their roles remain controversial. , Tumor necrosis factorlike weak inducer of apoptosis (TWEAK) has been suggested as a marker for CKD.  However, there are scanty data available regarding its role in CKD. The current study aimed at evaluating the levels of IL-2, IL-8 and TWEAK in HD and renal transplant patients as possible indicators of disease severity.
| Materials and Methods|| |
A total of 152 individuals participated in this study over a two-year period including 53 HD patients (preand post-dialysis), 45 renal transplant patients, and 54 healthy controls. HD patients were dialyzed for 3-4 h, two to three times weekly on Braun-dialysis machine (B. Braun Medical Inc., Michigan, USA) using biocompatible polysulfone membrane. Patients who had infectious diseases, immunological diseases, and malignancy were excluded from the study. Blood samples (5 mL) were collected in plain tubes. The samples were centrifuged with a speed of 3000 rpm at 4°C for 10 min to separate the serum from the blood cells. Then, the serum was pipetted and stored at −80°C till use.
Serum levels of IL-2, IL-8 and TWEAK were measured by enzyme-linked immunosorbent assay. These kits were brought from R&D Systems Europe (Abingdon, UK). The manufacturer's instructions were followed.
Ethical approval was obtained from the Medical Research Committee and Ethics Committee of Sultan Qaboos University, College of Medicine and Health Sciences (MREC# 538). All participants gave written consent.
| Statistical Analysis|| |
Statistical analysis was performed using SPSS (version 19.0) statistical package (SPSS Inc., Chicago, IL, USA). The numerical variables are presented as mean ± standard deviation. For data that did not follow normal distribution, Mann-Whitney U-test and Wilcoxon signedrank test (for paired difference between preand post-dialysis) were used for comparison, and P <0.05 was considered statistically significant.
| Results|| |
The HD group consisted of 53 patients (25 females and 28 males) with a mean age of 51 ± 13.6 years, whereas the renal transplant group consisted of 45 patients which included 27 stable transplants (60%) and 18 with renal biopsyproven rejections (40%) (14 females and 31 males) with a mean age of 41.1 ± 13.0 years and the healthy control group consisted of 54 individuals (22 females and 32 males) with a mean age of 40.73 ± 7.0 years.
Levels of IL-2, IL-8 and TWEAK cytokines were measured, and the mean concentration ± standard deviation was presented.
The mean concentration of serum IL-2 was significantly higher in HD and renal transplant samples when compared to the healthy samples. No significant difference was observed between preand post-dialysis samples [Figure 1].
|Figure 1: Serum IL-2 mean concentration (pg/mL) of healthy controls, pre-dialysis, post-dialysis and renal transplant patients. Results are expressed as mean ± SD.|
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The mean concentration of IL-8 was significantly higher in HD samples when compared to the healthy samples. No significant difference was seen between pre and post-dialysis samples and between renal transplant and the healthy samples [Figure 2].
|Figure 2: Serum IL-8 mean concentration (pg/mL) of healthy controls, pre-dialysis, post-dialysis and renal transplant patients. Results are expressed as mean ± SD.|
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The mean concentration of TWEAK was significantly higher in pre-dialysis samples when compared to the healthy and post-dialysis samples. However, TWEAK mean concentration was not significantly high in post-HD and renal transplant samples when compared to the healthy samples.
| Discussion|| |
The findings of this study show that IL-2 was significantly higher in dialysis patients compared to the healthy subjects. However, there was no significant change between preand postdialysis samples. Similar findings were also reported by others. , IL-2 is a large molecule measuring 15-18 kD that is produced by numerous types of immune cells including Band T-lymphocytes, natural killer cells, and dendritic cells. Being a large molecule, the removal of IL-2 during HD is low  compared to IL-6 and IL-10.  Different dialysis membranes can eliminate different cytokines, including IL-2.  In the current study, we used a polysulfone dialyzer, which has an excellent biocompatibility with minimal leukocyte reduction.
Some studies showed that IL-2 concentrations in HD patients are similar to those in the healthy controls. , The reason could be due to the impairment in activation and proliferation of T-cells to synthesize IL-2 in response to the increase in inflammatory cytokines such as tumor necrosis factor-α (TNF-α) and IL-6. , However, these findings are contradictory with other studies that showed that elevated levels of IL-2 in HD patients are associated with survival rate. 
We also observed high levels of IL-2 in renal transplant patients. Other studies have correlated high levels of IL-2 with improved survival for renal transplants. , In contrast, another study has linked high levels of IL-2 with renal rejections.  Elevated levels of IL-2 are usually associated with diseases such as rheumatoid arthritis, scleroderma, gastric cancer, small cell lung cancer, and hepatitis B infection. ,,,, In the current study, renal transplant patients are free of those diseases. Information on serum IL-2 in HD patients and healthy controls are scanty. In this study, we report low levels of IL-2 expression in healthy controls.
TWEAK is a member of TNF superfamily of cytokines. It has a molecular weight of 18 kD and is found in different organs such as kidney, liver, brain, ovary, and intestine. , It is a Type II transmembrane glycoprotein that induces several cell responses including cellular growth, proliferation, inflammation, and angiogenesis.  TWEAK is considered to be a recent marker in patients with CKD. , However, its role in HD and renal transplant patients is not yet clear as little information is available on this issue.
We found that TWEAK levels were high in pre-dialysis patients when compared with the healthy controls. This finding is not in agreement with another study which found that TWEAK levels were low in patients with CKD and high in healthy controls.  Similar findings were also reported. , However, it must be noted that these studies were conducted in nondialysis CKD patients classified according to the estimated glomerular filtration rate. In the current study, we divided the CKD patients into two groups, HD and those who had renal transplants.
We also observed that TWEAK levels in renal transplants were similar to those of healthy controls. The mechanism of TWEAK in CKD is not clear; one theory suggests that binding to its receptor, fibroblast growth factor-inducible 14, triggers many biological effects such as apoptosis, cellular growth, proliferation, and angiogenesis. , Further studies are needed to investigate the role of TWEAK in HD and renal transplant patients.
IL-8 is a member of the CXC chemokine family and secreted by many cells such as monocytes, T-cells, fibroblasts, endothelial cells, keratinocytes, hepatocytes, chondrocytes, neutrophils, and epithelial cells. The current study showed an increase in the levels of IL-8 in HD patients. This finding is in agreement with another similar study which found a significant increase in the serum levels of IL-8 in 37 patients who underwent HD.  Another study reported a significant increase of IL-8 levels in the dialysis fluid of patients with peritonitis undergoing continuous ambulatory peritoneal dialysis.  In fact, serum IL-8 could be used as an early biomarker of acute kidney injury, for example, it was found to be raised 2 h after cardiopulmonary bypass in patients who develop acute kidney injury.  Moreover, in animal models, increased serum IL-8 was used as an early marker for acute kidney injury.  The findings of this study showed that IL-8 levels remained unchanged after a HD session. This finding is not in line with another study which reported that IL-8 decreased after a HD session probably due to IL-8 diffusion through the dialysis membrane. 
We also observed that the levels of IL-8 in renal transplants were insignificantly higher compared with the healthy controls. This finding is in agreement with another study which showed no significant alteration of blood serum levels of IL-8 after renal transplantation.  As a weakness of this study, we should mention that the measurement of a panel of cytokines such as IL-1, IL-6, IL-10, IL-12, IL-12, IL-18, TNF-α, transforming growth factor-β as well as Creactive protein might provide more useful information about the status of HD and renal transplant patients.
| Conclusion|| |
The findings of this study showed high levels of IL-2, IL-8, and TWEAK in HD patients and normal levels of IL-8 and TWEAK and high levels of IL-2 in renal transplant patients.
Conflict of interest: None declared.
| References|| |
Genest J. C-reactive protein: Risk factor, biomarker and/or therapeutic target? Can J Cardiol 2010;26 Suppl A:41A-4A.
Shamseddin MK, Parfrey PS. Sudden cardiac death in chronic kidney disease: Epidemiology and prevention. Nat Rev Nephrol 2011;7:145-54.
Silverstein DM. Inflammation in chronic kidney disease: Role in the progression of renal and cardiovascular disease. Pediatr Nephrol 2009; 24:1445-52.
Oberg BP, McMenamin E, Lucas FL, et al. Increased prevalence of oxidant stress and inflammation in patients with moderate to severe chronic kidney disease. Kidney Int 2004;65:1009-16.
Tashiro K, Koyanagi I, Saitoh A, et al. Urinary levels of monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8), and renal injuries in patients with type 2 diabetic nephropathy. J Clin Lab Anal 2002;16:1-4.
Fallahzadeh MK, Roozbeh J, Geramizadeh B, Namazi MR. Interleukin-2 serum levels are elevated in patients with uremic pruritus: A novel finding with practical implications. Nephrol Dial Transplant 2011;26:3338-44.
Liu KD, Altmann C, Smits G, et al. Serum interleukin-6 and interleukin-8 are early biomarkers of acute kidney injury and predict prolonged mechanical ventilation in children undergoing cardiac surgery: A case-control study. Crit Care 2009;13:R104.
Yilmaz MI, Carrero JJ, Ortiz A, et al. Soluble TWEAK plasma levels as a novel biomarker of endothelial function in patients with chronic kidney disease. Clin J Am Soc Nephrol 2009; 4:1716-23.
Rysz J, Banach M, Cialkowska-Rysz A, et al. Blood serum levels of IL-2, IL-6, IL-8, TNFalpha and IL-1beta in patients on maintenance hemodialysis. Cell Mol Immunol 2006;3:151-4.
Tarakçioglu M, Erbagci AB, Usalan C, Deveci R, Kocabas R. Acute effect of hemodialysis on serum levels of the proinflammatory cytokines. Mediators Inflamm 2003;12:15-9.
Rostaing L, Peres C, Tkaczuk J, et al. Ex vivo flow cytometry determination of intracytoplasmic expression of IL-2, IL-6, IFN-gamma, and TNF-alpha in monocytes and T lymphocytes, in chronic hemodialysis patients. Am J Nephrol 2000;20:18-26.
van Riemsdijk-Van Overbeeke IC, Baan CC, Knoop CJ, Loonen EH, Zietse R, Weimar W. Quantitative flow cytometry shows activation of the TNF-alpha system but not of the IL-2 system at the single cell level in renal replacement therapy. Nephrol Dial Transplant 2001; 16:1430-5.
Kimmel PL, Phillips TM, Simmens SJ, et al. Immunologic function and survival in hemodialysis patients. Kidney Int 1998;54:236-44.
Amirzargar A, Lessanpezeshki M, Fathi A, et al. TH1/TH2 cytokine analysis in Iranian renal transplant recipients. Transplant Proc 2005;37: 2985-7.
Orditura M, Romano C, De Vita F, et al. Behaviour of interleukin-2 serum levels in advanced non-small-cell lung cancer patients: Relationship with response to therapy and survival. Cancer Immunol Immunother 2000; 49:530-6.
Forones NM, Mandowsky SV, Lourenço LG. Serum levels of interleukin-2 and tumor necrosis factor-alpha correlate to tumor progression in gastric cancer. Hepatogastroenterology 2001; 48:1199-201.
Kahaleh MB, LeRoy EC. Interleukin-2 in scleroderma: Correlation of serum level with extent of skin involvement and disease duration. Ann Intern Med 1989;110:446-50.
Tebib JG, Boughaba H, Letroublon MC, et al. Serum IL-2 level in rheumatoid arthritis: Correlation with joint destruction and disease progression. Eur Cytokine Netw 1991;2:239-43.
Bozkaya H, Bozdayi M, Türkyilmaz R, et al. Circulating IL-2, IL-10 and TNF-alpha in chronic hepatitis B: Their relations to HBeAg status and the activity of liver disease. Hepatogastroenterology 2000;47:1675-9.
Ortega L, Fornoni A. Role of cytokines in the pathogenesis of acute and chronic kidney disease, glomerulonephritis, and end-stage kidney disease. Int J Interferon Cytokine Mediat Res 2010;2:49-62.
Justo P, Sanz AB, Sanchez-Niño MD, et al. Cytokine cooperation in renal tubular cell injury: The role of TWEAK. Kidney Int 2006; 70:1750-8.
Carrero JJ, Ortiz A, Qureshi AR, et al. Additive effects of soluble TWEAK and inflammation on mortality in hemodialysis patients. Clin J Am Soc Nephrol 2009;4:110-8.
Hassan SB, El-demery AB, Ahmed AI, Abukhalil RE. Soluble TWEAK and cardiovascular morbidity and mortality in chronic kidney disease patients. Arab J Nephrol Transplant 2012;5:27-32.
Yilmaz MI, Sonmez A, Ortiz A, et al. Soluble TWEAK and PTX3 in nondialysis CKD patients: Impact on endothelial dysfunction and cardiovascular outcomes. Clin J Am Soc Nephrol 2011;6:785-92.
Meier P. Plasma sTWEAK and PTX3: New determinant tools of cardiovascular outcome also in patients with CKD. Clin J Am Soc Nephrol 2011;6:697-9.
Polek TC, Talpaz M, Darnay BG, SpivakKroizman T. TWEAK mediates signal transduction and differentiation of RAW264.7 cells in the absence of Fn14/TweakR. Evidence for a second TWEAK receptor. J Biol Chem 2003; 278:32317-23.
Ho DH, Vu H, Brown SA, Donohue PJ, Hanscom HN, Winkles JA. Soluble tumor necrosis factor-like weak inducer of apoptosis overexpression in HEK293 cells promotes tumor growth and angiogenesis in athymic nude mice. Cancer Res 2004;64:8968-72.
Nakanishi I, Moutabarrik A, Okada N, et al. Interleukin-8 in chronic renal failure and dialysis patients. Nephrol Dial Transplant 1994; 9:1435-42.
Ko YC, Mukaida N, Kasahara T, et al. Specific increase in interleukin-8 concentrations in dialysis fluid of patients with peritonitis receiving continuous ambulatory peritoneal dialysis. J Clin Pathol 1995;48:115-9.
Hoke TS, Douglas IS, Klein CL, et al. Acute renal failure after bilateral nephrectomy is associated with cytokine-mediated pulmonary injury. J Am Soc Nephrol 2007;18:155-64.
Rysz J, Kocur E, Blaszczak R, Bartnicki P, Stolarek RA, Piechota M. IL-2, IL-6 and IL-8 levels remain unaltered in the course of immunosuppressive therapy after renal transplantation. Cent Eur J Med 2008;3:199-202.
Department of Allied Health Sciences, College of Medicine and Health Sciences, Sultan Qaboos University, P. O. Box 35, Postal Code 123, Muscat
Sultanate of Oman
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