Annals of Tropical Medicine and Public Health

ORIGINAL ARTICLE
Year
: 2017  |  Volume : 10  |  Issue : 6  |  Page : 1656--1660

Analysis of the association between IL-23R rs11209026 polymorphism in rheumatoid arthritis and systemic lupus erythematosus in an Iranian population


Shadi Soleymani Motlagh1, Mahmood Deghani Ashkezari1, Kolsum Ahmadi2, Azam Soleymani3, Ali Asghar Kiani4,  
1 Medical Biotechnology Research Center, Ashkezar Branch, Islamic Azad University, Ashkezar, Yazd, Iran
2 Student Research Committee, Lorestan University of Medical Sciences, Khorramabad, Iran
3 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran
4 Razi Herbal Medicines Research Center and School of Allied Medical Sciences, Department of Hematology and Blood Transfusion, Lorestan University of Medical Sciences, Khorramabad, Iran

Correspondence Address:
Ali Asghar Kiani
Department of Hematology and Blood Transfusion, Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad
Iran

Abstract

Introduction: Rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE) are two autoimmune inflammatory diseases with possible genetic links. In this study, the polymorphism R381Q IL-23R in patients with RA, SLE, and healthy controls and its relation to genotype was investigated. Materials and Methods: In this case-control study of 100 patients with SLE, 100 patients with RA and 112 healthy individuals who were referred to the rheumatology ward of some hospitals in Khorramabad (Lorestan province, west of Iran) were selected. Genomic DNA was extracted from peripheral blood and genotyping was performed using polymerase chain reaction-restriction fragment length polymorphism method. Fisher's exact test was used to compare data from genotypes and alleles between all groups. Results: It was found that polymorphism A > G rs11209026 of IL-23R gene in the control group and patients with RA had significant difference (P = 0.046). However in the control group and SLE patients, there was no significant difference (P = 0.51). Conclusion: The results indicate that single nucleotide polymorphism rs11209026 G > A of the IL-23R gene is possibly associated with RA. IL-23 is an important inflammatory cytokine and essential for the differentiation of Th17 cells. Hence studies related to the role of Th17 cells in the pathogenesis of RA would help provide more insights into its regulatory mechanisms.



How to cite this article:
Motlagh SS, Ashkezari MD, Ahmadi K, Soleymani A, Kiani AA. Analysis of the association between IL-23R rs11209026 polymorphism in rheumatoid arthritis and systemic lupus erythematosus in an Iranian population.Ann Trop Med Public Health 2017;10:1656-1660


How to cite this URL:
Motlagh SS, Ashkezari MD, Ahmadi K, Soleymani A, Kiani AA. Analysis of the association between IL-23R rs11209026 polymorphism in rheumatoid arthritis and systemic lupus erythematosus in an Iranian population. Ann Trop Med Public Health [serial online] 2017 [cited 2019 Dec 9 ];10:1656-1660
Available from: http://www.atmph.org/text.asp?2017/10/6/1656/222691


Full Text



 Introduction



Rheumatoid arthritis (RA) is a systemic autoimmune disease identified by erosive chronic polyarthritis, progressive joint inflammation, and destruction of articular cartilage.[1],[2] The chronic inflammation process associated with autoantibodies production, formation, and deposition of immune complexes in the joint is responsible for stimulating destructive mechanisms which cause structural damage and lead to joint dysfunction and disability.[3],[4]

Systemic lupus erythematosus (SLE) is an other autoimmune disease that is determined by the production of antibodies against components of the cell nucleus generating specific clinical and immunological complications.[5] The exact etiology of SLE is unknown but pathological findings in many patients are related to inflammation, vasculitis, immune complex deposition, and vasculopathy.[6] Cytokines play a major role in the processes that cause inflammation, articular destruction and the pathological manifestations associated with RA and SLE.[7] IL-23 is one of the important cytokines which belong to the type I cytokines superfamily. This cytokine is secreted by macrophages and promote expansion of Th17 cells from naive CD4 + T cells.[8] CD4+ T cells- derived Th17 have been involved in autoimmune tissue inflammation by producing of IL-17 cytokin. In addition Th17 produce other cytolkines such as IL-6, IL-22, TNF-a, GM-CSF that is involved in inflamation process.[9] Due to the expression of IL-23R in Th17 cells which results in the induction of Th17 cells, IL-17 is produced; therefore, targeting the IL-17/23 axis (as an inflammatory axis) may be a new therapeutic strategy for many autoimmune inflammatory diseases.[10] Regarding the role of IL-23 in Th17 cells development, it seems plausible that genetic variations in IL-23R would influence the cytokine profile of Th17 cells.

In recent years, several studies have suggested R381Q IL23 (IL23R) variants as a predisposing factor in inflammatory diseases.[11]

The aim of this study was to determine the relationship between R381Q IL-23 receptor polymorphism in patients with RA and SLE.

 Materials and Methods



Participants

In this case–control study a total of 200 patients including 100 patients with RA and 100 patients with SLE and 112 healthy controls referred to rheumatology ward of Khorramabad (Iran) Laboratory. Patients were selected based on clinical test and diagnosed with Rheumatologist. All patients and healthy individuals writed the consent and content of study was approved by Ethics Committee of ISLAMIC AZAD UNIVERSITY, Ashkezar Branch (Yazd, Iran). DNA extraction from whole blood samples was performed by Genomic DNA purification kit (Roche, Germany). Yields of DNA were evaluated from both qualitative and quantitative with spectrophotometry and 1% agarose gel electrophoresis, respectively. To genotype, Arg381Gln polymorphism polymerase chain reaction was applied and a PCR-restriction fragment length polymorphism (RFLP) method was used to investigation of single nucleotide polymorphisms in the Arg381Gln IL-23R variant. A pair of oligonucleotide primers were obtained of GenBank and were synthesized to the PCR process. The primer sequences included in study are as follows 50-CTTTTCTGGCAGGGTCATTTTG-30 (sense) and 50-AAGTTGTTTCCTGGGGTAGTTGTG-30 (antisense).[12] The PCR was conducted in a volume of 25 μl containing Master mixes (1X) prepared in 12.5 μl, 2 μl of each primer and 2 μl DNA at a primer annealing temperature of 50 °C. The amplified product was run on a 1% agarose gel and stained with safe stain for observation on an ultraviolet (UV) transilluminator. Amount of,10 μl of amplicons was treated with 0.2 μl of Hpy188I restriction endonuclease (New England Biolabs, MA, USA) under the one hour at 37 °C condition. The digestion products were then run on a 2% agarose gel and stained with safe stain for visualization on a UV transilluminator. Hpy 188I digestion of homozygous normal DNA fragments of 288, 103, 82 and 35 base pairs. In addition, DNA containing the heterozygous mutant fragments of 323, 103 and 82 base pairs as shown in [Figure 1].{Figure 1}

Statistical analysis

Data analysis was performed by Hardy–Weinberg equilibrium and chi-squared tests. Data analysis of genotypes and alleles compared with the control group was used by Fisher's exact test. P values <0.05 were deemed statistically significant in all subjects.

 Results



This study evaluated 100 patients with RA, 71 patients (71%) were female and 29 (29%) were male and 100 patients with SLE, 68 patients (68%) were female and 32 (32%) were male. In the control group of 112 patients, 79 (53.70%) were female and 33 (47.29%) were male. Mean age ± standard deviation of participants during diagnosis was 46.19 ± 10.28 years, 44.91 ± 11.31 years, and 45.03 ± 10.36 years for RA, SLE, and control group, respectively. Age and sex were similar in both the patients and healthy control group and hence that the three groups were not statistically significant. Genotype distributions of SNP of IL-2R and allele frequencies are presented in [Figure 2] and [Figure 3], respectively. The results of Chi-square test indicated a significant difference in the presence of this genetic variant in RA patients and healthy controls which were 10.05% and 21.46%, respectively. In addition, the allelic frequency 1142A in patients with RA compared with the control group was statistically significant (7 and 30, respectively, (P P = 0.283) and allelic frequency (24 and 30, respectively) in SLE patients compared with controls when analyzed separately (P = 0.283).{Figure 2}{Figure 3}

 Discussion



It is found t significant association between A > G rs11209026 polymorphism of the IL-23R gene and RA (P = 0.046). However, there is not found any statistically significant in patients with SLE and the control group. Wild and mutant alleles were G and A, respectively. The allele frequency of mutant allele A was lower in the present study. In recent years, many studies have been conducted to prove the role of inflammation in RA. One important inflammatory pathway that has been considered is IL-23/Th-17 inflammatory axis.[13]

Our study was consistent with Geremia and Jewell (2012) that they showed the presence of SNP R381Q in patients with inflammatory bowel disease affected on IL-23 receptor signal transduction by reduce activity.[14] Capon et al. found that 381 Q allelic frequency in patients with the psoriasis disease was 3.6%, whereas in control subjects (healthy) was 7%.[15] Since they observed the higher frequency of 381 Q in healthy control, they suggested that SNP R381Q may play a protective role in autoimmune diseases and inflammation.[15] In addition in a study by Hazlett et al. demonstrated severity of RA disease had significant correlation with plasma IL-17 levels.[16] IL-23 regulates the functional responses of Th17 cells including differentiation and cell survival. Since in contrast to naive T cells, IL-23R frequently expressed on activated T cells. Thus, In the absence of IL-23, Th17 cells do not able to qualified activated T cells, even if other essential cytokines for activation of effector T cells are also present. It thought that IL-23R gene polymorphism affected on the Th17/IL-23axis. IL-23 can induce signal transduction by binding its receptor as IL-23 R.[13] IL-23R is a complex formed by two heterogonous subunits including IL-12RB1 that is shared among different cytokine receptor and the JAK kinases related receptor seems to be a nonspecific subunit.[17] Phosphorylation of tyrosine residues in Jak2 subunits of the intracellular domain of the IL-23R subunit triggers by binding of IL-23 to its receptor. After tyrosine Phosphorylation STAT3 molecules can form complex and be Phosphorylated.[18] STAT3 proteins after Phosphorylation induce the transcription of several cytokines such as IL-17, IL-22 and interferon in the nucleus.[19] R381Q polymorphism is preserved among different species and located in trans membrane domain of IL-23 receptor. The R381Q IL-23R variant may affect the IL-23R signal transduction component and may control phosphorylation and transcriptional activation.[20],[21],[22],[23],[24] In accordance with this, Sarinet al observed that decreased production of IL-22 and IL-17is simultaneously associated with the significant correlation between R381Q and reduced expression of STAT3. They concluded that the protection role of R381Q in autoimmunity disease.[19] Gui-Mei Chenet al. indicated that IL-23R gene polymorphisms were not associated with SLE.[5] However, it is reported the IL-23R Arg381Gln polymorphism has been associated with RA. We can conclude the presences of GG genotype causes a specific conformational of IL-23 protein receptors that lead to a stronger signal transduction than the AG genotype

 Conclusion



RA is known as a multifactorial disease and many factors such as genetics, environment and their interactions that are involved in disease. The study indicates that the polymorphismrs11209026 G > A SNP of IL-23R gene may have a correlation with the incidence of RA.[23] However, whether there is a biologic underlying mechanism (e.g., a sustained malfunction of Th17 cells) dFurther mechanistic investigations are needed to actual determination of functions of IL-23R gene variants in the immunopathogenesis of RA and SLE.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

References

1Chan PS, Kong KO. Natural history and imaging of subtalar and midfoot joint disease in rheumatoid arthritis. Int J Rheum Dis 2013;16:14-8.
2van Boekel MA, Vossenaar ER, van den Hoogen FH, van Venrooij WJ. Autoantibody systems in rheumatoid arthritis: Specificity, sensitivity and diagnostic value. Arthritis Res 2002;4:87-93.
3Majithia V, Geraci SA. Rheumatoid arthritis: Diagnosis and management. Am J Med 2007;120:936-9.
4Michou L, Teixeira VH, Pierlot C, Lasbleiz S, Bardin T, Dieudé P, et al. Associations between genetic factors, tobacco smoking and autoantibodies in familial and sporadic rheumatoid arthritis. Ann Rheum Dis 2008;67:466-70.
5Chen GM, Feng CC, Ye QL, Wang J, Cen H, Li R, et al. Lack of association between IL-23R gene polymorphisms and systemic lupus erythematosus in a chinese population. Inflamm Res 2013;62:791-5.
6Jakes RW, Bae SC, Louthrenoo W, Mok CC, Navarra SV, Kwon N, et al. Systematic review of the epidemiology of systemic lupus erythematosus in the Asia-Pacific region: Prevalence, incidence, clinical features, and mortality. Arthritis Care Res (Hoboken) 2012;64:159-68.
7Veldhoen M, Hocking RJ, Atkins CJ, Locksley RM, Stockinger B. TGFbeta in the context of an inflammatory cytokine milieu supports de novo differentiation of IL-17-producing T cells. Immunity 2006;24:179-89.
8Duerr RH, Taylor KD, Brant SR, Rioux JD, Silverberg MS, Daly MJ, et al. A  genome-wide association study identifies IL23R as an inflammatory bowel disease gene. Science 2006;314:1461-3.
9Zhu S, Qian Y. IL-17/IL-17 receptor system in autoimmune disease: Mechanisms and therapeutic potential. Clin Sci (Lond) 2012;122:487-511.
10Isailovic N, Daigo K, Mantovani A, Selmi C. Interleukin-17 and innate immunity in infections and chronic inflammation. J Autoimmun 2015;60:1-1.
11Sato K, Suematsu A, Okamoto K, Yamaguchi A, Morishita Y, Kadono Y, et al. Th17 functions as an osteoclastogenic helper T cell subset that links T cell activation and bone destruction. J Exp Med 2006;203:2673-82.
12Venegas M, Beltrán CJ, Alvarez L, Castro A, Torres T, Leal AD, et al. IL-23R arg381Gln polymorphism in Chilean patients with inflammatory bowel disease. Eur Cytokine Netw 2008;19:190-5.
13Wong CK, Lit LC, Tam LS, Li EK, Wong PT, Lam CW, et al. Hyperproduction of IL-23 and IL-17 in patients with systemic lupus erythematosus: Implications for xTh17-mediated inflammation in auto-immunity. Clin Immunol 2008;127:385-93.
14Geremia A, Jewell DP. The IL-23/IL-17 pathway in inflammatory bowel disease. Expert Rev Gastroenterol Hepatol 2012;6:223-37.
15Capon F, Di Meglio P, Szaub J, Prescott NJ, Dunster C, Baumber L, et al. Sequence variants in the genes for the interleukin-23 receptor (IL23R) and its ligand (IL12B) confer protection against psoriasis. Hum Genet 2007;122:201-6.
16Hazlett J, Stamp LK, Merriman T, Highton J, Hessian PA. IL-23R rs11209026 polymorphism modulates IL-17A expression in patients with rheumatoid arthritis. Genes Immun 2012;13:282-7.
17Di Meglio P, Di Cesare A, Laggner U, Chu CC, Napolitano L, Villanova F, et al. The IL23R R381Q gene variant protects against immune-mediated diseases by impairing IL-23-induced th17 effector response in humans. PLoS One 2011;6:e17160.
18Abdollahi E, Tavasolian F, Momtazi-Borojeni AA, Samadi M, Rafatpanah H. Protective role of R381Q (rs11209026) polymorphism in IL-23R gene in immune-mediated diseases: A comprehensive review. J Immunotoxicol 2016;13:286-300.
19Sarin R, Wu X, Abraham C. Inflammatory disease protective R381Q IL23 receptor polymorphism results in decreased primary CD4+and CD8+human T-cell functional responses. Proc Natl Acad Sci U S A 2011;108:9560-5.
20Gaston JS, Goodall JC, Baeten D. Interleukin-23: A central cytokine in the pathogenesis of spondylarthritis. Arthritis Rheum 2011;63:3668-71.
21Pidasheva S, Trifari S, Phillips A, Hackney JA, Ma Y, Smith A, et al. Functional studies on the IBD susceptibility gene IL23R implicate reduced receptor function in the protective genetic variant R381Q. PLoS One 2011;6:e25038.
22Abdi J, Chen G, Chang H. Drug resistance in multiple myeloma: Latest findings and new concepts on molecular mechanisms. Oncotarget 2013;4:2186-207.
23Rueda B, Orozco G, Raya E, Fernandez-Sueiro JL, Mulero J, Blanco FJ, et al. The IL23R arg381Gln non-synonymous polymorphism confers susceptibility to ankylosing spondylitis. Ann Rheum Dis 2008;67:1451-4.
24Soltanpour MS, Soheili Z, Pourfathollah AA, Samiei S, Meshkani R, Kiani AA et al. The A1298C mutation in methylenetetrahydrofolate reductase gene and its association with idiopathic venous thrombosis in an Iranian population. Lab Medicine 2011;42:213-6.