Nucleotide Polymorphisms Of Cytokine Genes Research Articles

Single nucleotide polymorphisms in cytokine genes and their association with primary Sjögren's syndrome in Saudi patients: A cross-sectional study.

To determine the allelic frequencies and effects of genotypic variations in cytokine gene polymorphisms in a Saudi Arabian population. This cross-sectional study involved 41 patients with Primary Sjögren's syndrome (pSS) and 71 healthy controls between October 2018 and May 2019. Single nucleotide polymorphisms genotyping was performed using the SEQUENOM MassARRAY® System, targeting nine polymorphisms in different cytokine genes. Chi-square tests were used to compare the patients and controls. The interleukin-1 beta (IL-1β) rs1143627 CT (control, 52.7%; patients, 21.2%) and TT + CT (p= 0.003; p=0.033) genotypes were less frequent in patients with pSS than in healthy controls. The C allele in rs10488631 in the interferon regulatory factor 5 (IRF5) gene and the A allele in rs12583006 in the B-cell activating factor (BAFF) gene were associated with an increased risk of pSS development in the patient group. The CT genotype at -31 (rs1143627) in the IL-1β gene was not associated with a high risk of pSS development in the Saudi population, in contrast to what has been verified in other ethnicities. However, the C allele in rs10488631 in IRF-5 and the A allele in rs12583006 in BAFF were associated.

Association of Single Nucleotide Polymorphisms of Cytokine Genes with Depression, Schizophrenia and Bipolar Disorder.

Immune gene variants are known to be associated with the risk of psychiatric disorders, their clinical manifestations, and their response to therapy. This narrative review summarizes the current literature over the past decade on the association of polymorphic variants of cytokine genes with risk, severity, and response to treatment for severe mental disorders such as bipolar disorder, depression, and schizophrenia. A search of literature in databases was carried out using keywords related to depressive disorder, bipolar disorder, schizophrenia, inflammation, and cytokines. Gene lists were extracted from publications to identify common genes and pathways for these mental disorders. Associations between polymorphic variants of the IL1B, IL6, and TNFA genes were the most replicated and relevant in depression. Polymorphic variants of the IL1B, IL6, IL6R, IL10, IL17A, and TNFA genes have been associated with schizophrenia. Bipolar disorder has mainly been associated with polymorphic variants of the IL1B gene. Interestingly, the IL6R gene polymorphism (rs2228145) was associated with all three diseases. Some cytokine genes have also been associated with clinical presentation and response to pharmacotherapy. There is also evidence that some specific polymorphic variants may affect the expression of cytokine genes. Thus, the data from this review indicate a link between neuroinflammation and severe mental disorders.

Single Nucleotide Polymorphisms in Cytokine Genes are Associated with the Susceptibility to Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is the most common type of cancer among men in the Indian subcontinent. Cytokines regulate inflammation and angiogenesis in a variety of cancers. Genetic variability in the cytokine genes can potentially influence the predisposition to oral carcinogenesis. The aim of the current study was to investigate the associations of SNPs in cytokine genes with the susceptibility of oral squamous cell carcinoma. In the present study, we have analyzed the allelic frequency of 32 single nucleotide polymorphisms (SNPs) using MassArray-based iPLEX assay in 16 cytokine genes in 166 OSCC patients and 151 healthy subjects from central India. Out of 32 SNPs analyzed, five SNPs were significantly associated with the risk of OSCC. AA and GG genotypes of IL-1β +3953 were associated with an increased and decreased risk of OSCC, respectively. In several genetic models, GG genotype and G allele in IL-12A 3'UTR G>A were found to be associated with an increased risk of OSCC. Similarly, the GG genotype of IL-12B +1188 T>G was associated with increased susceptibility to OSCC. We conclude that SNPs in the genes coding for IL-1β, IL-12A and IL-12B are associated with increased genetic susceptibility to OSCC in the central Indian population.

CHARACTERISTICS OF CYTOKINE GENE POLYMORPHISMS IN CHILDREN WITH DIFFERENT PHENOTYPES OF BRONCHIAL ASTHMA.

Abstract
 BACKGROUND: Bronchial asthma is a chronic inflammatory disease of the airways, the development of which is based on genetic predictors associated with the differentiation and functioning of T-helpers. Polymorphisms in the genes of cytokines involved in the regulation of the direction of the T-helper mediated immune response are risk factors for the development of the disease and the realization of various phenotypes of bronchial asthma.
 AIMS: Study of the structure and frequency of occurrence of single nucleotide polymorphisms of cytokine genes with an assessment of the risk of various phenotypes of bronchial asthma in children.
 MATERIALS AND METHODS: In this case control study, 250 children were examined, including 150 children with a verified diagnosis of bronchial asthma (including 75 children with virus-induced and 75 children with allergen-induced disease phenotypes) and 100 sexually comparable healthy peers. The children underwent a comprehensive general clinical and allergological examination, genotyping, analysis of the structure, frequency of occurrence of cytokine gene polymorphisms and calculation of the odds ratio of the risk of developing different bronchial asthma phenotypes. DNA samples isolated from peripheral venous blood were used as material for molecular genetic analysis. The following mutation points were selected: IFN (T-874 A), IL-4 (C-589 T), IL-6 (C-174 G), IL-17A (G-197 A), TNF (G-308 A).
 The work followed the ethical principles set forth by the Declaration of Helsinki by the World Medical Association and the Rules of Clinical Practice in the Russian Federation, the study design was approved by the Interdisciplinary Ethics Committee of the Federal State Budgetary Educational Institution of Higher Education Pacific State Medical University of the Ministry of Healthcare of the Russian Federation on April 27, 2015 (Protocol No. 8), informed voluntary consent was signed by the parents.
 When processing digital data, we used the methods of descriptive, parametric and nonparametric statistics of the Statistica 10 program, comparison of unrelated groups by qualitative characteristics, assessment of the correspondence of the distributions of genotypes to the expected values at the Hardy-Weinberg equilibrium. The analysis of the frequency distributions of genotypes and alleles in two subpopulations was carried out using the Chi-square test (2).
 RESULTS: A comparative analysis of the frequencies of alleles and genotypes of cytokines of various Th profiles with the definition of features in allergen-induced and virus-induced phenotypes of the disease revealed in children with bronchial asthma the predominance of homozygous genotypes IFN (A-874A), IL-4 (T-589T), IL- 6 (G-174G), IL-17A (A-197A), TNF (A-308A), and in healthy peers the prevalence is IFN (T-874T), IL-4 (C-589C), IL-6 (C -174C), IL-17A (G-197G), TNF G-308G. Heterozygous genotypes IL-4 (C -589T), IL-6 (G -174C), IL-17A (G -197A), TNF (G -308A) found in children with bronchial asthma more often than in healthy peers, with the exception of the IFN genotype (T -874A). In children with the virus-induced bronchial asthma phenotype, the presence of the IL-4 (C-589T) mutant allele was found in 30,67% of cases with an odds ratio of -19,3 CI 95%. (11,23-33,31). When carrying the mutant A genotype IFN (T-874A), the odds ratio of the risk of developing the disease reflected a high degree of probability of the virus-induced bronchial asthma phenotype (OR = 5,11 (3,18-8,23) CI 95%). Carriage of homozygous genotypes IL-6 (G-174G), IL-17A (A-197A) determined an increased risk of developing allergen-induced bronchial asthma (OR = 2,71 (1,73-4,18) CI 95% and OR = 0,51 (0,32-0,71) CI 95%, respectively). Among children with bronchial asthma, there was a statistically significant increase in the incidence of the functionally unfavorable genotype 308 A /A of the TNF 308G /A gene, and the odds ratio reflects a 2,6-fold increase in the risk of developing a virus-induced bronchial asthma phenotype (2 = 18,66; p = 0,017; OR = 2,60, CI 95% (1,67-4,01).
 CONCLUSIONS: As a result of the study, significant differences were determined in the structure and frequency of occurrence of cytokine gene polymorphisms in children with allergen and virus-induced bronchial asthma, depending on the realized phenotype of the disease. Carriage of mutant alleles IFN (A-874A), IL-4 (T-589T), IL-6 (G-174G), IL-17A (A-197A), TNF (A-308A) can be characterized as genetic predictors of bronchial asthma, for the implementation of the virus-induced phenotype, the odds ratio is higher in the presence of mutant alleles IFN (A-874A), IL-4 (T-589T), TNF (A-308A), for the allergen-induced phenotype of the disease - IL-6 (G -174G), IL-17A (A-197A).

Single Nucleotide Polymorphisms of Cytokine Genes in Warm Autoimmune Hemolytic Anemias: Relationship with Clinical and Hematological Parameters

Background: Autoimmune hemolytic anemia (AIHA) is due to an increased immune peripheral destruction of red blood cells. The disease is characterized by high heterogeneity regarding anemia severity, treatment requirement and occurrence of complications. It has been shown that Hb levels at baseline, presence of complement activation, and the degree of bone marrow compensation are the main predictors of outcome. Moreover, multi-relapsing cases show bone marrow dysmyelopoiesis, and fibrosis suggesting an immune attack against bone marrow precursors. An interesting area to investigate is the genetic landscape of AIHA, including cytokine gene signature and other molecular mechanisms of refractoriness.Aim: To evaluate the frequency of cytokine single nucleotide polymorphisms (SNPs) in warm AIHA and their correlation with clinical and hematological parameters, including the degree of anemia, treatment refractoriness and number of therapies lines.Methods: We investigated SNPs of five cytokine genes [tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, transforming growth factor (TGF)-beta, interleukin (IL)-6 and IL-10] in 72 Italian warm AIHA patients, using the Cytokine Tray (One Lambda, Canoga Park, CA) according to the manufacturer's recommendations. Allele and genotype frequency results were compared to those described in the Italian population by Uboldi de Capei et al. (2003) and Poli et al. (2002).Results: Seventy-two warm AIHA patients have been evaluated: median age 63 years (range 17-90), classified as per DAT positivity in IgG+ (N=48), IgG+C (N=17), IgA+ (N=1), and DAT negative AIHA (N=6). Demographic and hematological characteristics of patients are described in Figure 1A. Regarding cytokine SNP frequencies, we observed that AIHA patients had a significantly higher prevalence of the TNF-alpha-308 high producers genotypes (G/G and G/A) compared to controls (40.3% vs 16% or 23%, p=0.0003 and p=0.006, respectively). No differences were detected regarding genotype frequencies of the other cytokine SNPs (Figure 1B). By dividing patients according to anemia severity at onset, a more frequent low producer genotype of IFN-gamma (A/A) was observed in patients with Hb levels >7.4 g/dL (median value of patients cohort) compared to the other group (p=0.05). Finally, considering the number of relapses or therapy lines, an increased frequency of TNF-alpha -308 high producers genotypes were found among multi-relapsing (> 3 relapses, 70.5% vs 56%) and multi-treated patients (> 3therapy lines, 52% vs 35%), although not significantly.Conclusions: These preliminary data show that wAIHA patients display a more pro-inflammatory cytokine signature at molecular level as compared to the Italian control population. This is highlighted by an increased prevalence of TNF-alpha high producer genotype that is also associated with multiple relapses and greater treatment requirement. Additionally, cytokine SNPs may be also associated with anemia severity at onset, as observed for IFN-gamma low producer genotype that was more frequent in less severe cases. [Display omitted] DisclosuresFattizzo: Alexion, Amgen, Annexon, Apellis, Grifols, Novartis, Momenta: Consultancy, Speakers Bureau. Bianchi: Agios pharmaceutics: Consultancy, Membership on an entity's Board of Directors or advisory committees. Barcellini: Agios: Honoraria, Research Funding; Bioverativ: Membership on an entity's Board of Directors or advisory committees; Novartis: Honoraria; Alexion Pharmaceuticals: Honoraria; Incyte: Membership on an entity's Board of Directors or advisory committees.

Analysis of the frequency of single nucleotide polymorphisms in cytokine genes in patients with New Onset Diabetes After Transplant

New Onset Diabetes After Transplantation (NODAT) is a serious metabolic complication. While β-cell dysfunction is considered the main contributing factor in the development of NODAT, the precise pathogenesis is not well understood. Cytokines are thought to be involved in the inflammation of islet β-cells in diabetes; however, few studies have investigated this hypothesis in NODAT. A total of 309 kidney transplant recipients (KTRs) were included in this study. An association between kidney transplants, and the development of diabetes after transplant (NODAT) was investigated. Comparison was made between KTRs who develop diabetes (NODAT cases) or did not develop diabetes (control), using key cytokines, IL-6 G (− 174)C, macrophage mediator; IL-4 C (− 490)T, T helper (Th)-2 cytokine profile initiator; Th-1 cytokine profile initiator interferon-γ T (+ 874) A gene and TGF β1 C (+ 869) T gene polymorphisms were investigated. The genes were amplified using well-established polymerase chain reaction (PCR) techniques in our laboratory. Compared to the AA and AT genotypes of interferon gamma (IFNG), there was a strong association between the TT genotype of IFNG and NODAT kidney transplant recipients (KTRs) versus non-NODAT KTRs (p = 0.005). The AA genotype of IFNG was found to be predominant in the control group (p = 0.004). Also, significant variations of IL6 G (− 174) C, IL-4 C (− 590) T, interferon-γ T (+ 874) A gene and transforming growth factor β1 C (+ 869) T may contribute to NODAT. Our data is consistent with theTh-1/T-reg pathway of immunity. Further larger pan Arab studies are required to confirm our findings.

Clinical implications of IL-10 promoter polymorphisms on disease susceptibility in Indian SLE patients.

Single nucleotide polymorphisms in cytokine genes including interleukin-10 have been described to play a vital role in the overall pathogenesis of systemic lupus erythematosus. However, due to a lack of evidence from the Indian population, this study was conducted to analyse the possible influence of interleukin-10 promoter polymorphisms over the disease susceptibility, serum interleukin-10 level and clinical manifestations of the disease in Indian systemic lupus erythematosus patients. In total, 200 systemic lupus erythematosus patients and 201 controls were recruited under this study. Genotyping of interleukin-10 (−1082A/G; −819T/C and −592C/A) polymorphisms was done by direct DNA sequencing and polymerase chain reaction-restriction fragment length polymorphism methods respectively. Serum interleukin-10 levels were measured by multiplex assay. Interleukin-10 −1082G and −592A allele frequencies were significantly increased in systemic lupus erythematosus patients (corrected p value <0.05). Also, combined −1082AG+GG, −819TC+CC and −592CA+AA genotype frequencies were significantly increased in the patient group. A higher trend of association between −1082AG+GG genotype frequency was observed in patients with serositis (odds ratio = 2.7, p = 0.0233, corrected p value = 0.2097). Serum interleukin-10 levels were significantly higher in systemic lupus erythematosus patients (4.3 ± 3.1 pg/ml) than controls (2.6 ± 1.4 pg/ml) ( p < 0.0001). Furthermore, interleukin-10 levels were positively correlated with disease activity ( p = 0.39, p < 0.0001). The frequency of the GCA (−1082, −819, −592) haplotype was significantly higher in systemic lupus erythematosus patients (10.6%) than controls (1.6%) (odds ratio = 5.4, p = 0.0330). Moreover, ACC, GCC and GCA haplotypes were found to be strongly associated with serositis. However, the frequency of the ACC haplotype was significantly higher in patients with neurologic involvement (odds ratio = 14.9, corrected p value <0.001). Thus, interleukin-10 promoter polymorphisms suggest they have a proactive role in increased susceptibility to the disease among Indian systemic lupus erythematosus patients.

Associations of common IL-4 gene polymorphisms with cancer risk: A meta-analysis.

Cancer incidence is dramatically increasing worldwide, therefore improved prediction and therapeutic methods are needed. Single nucleotide polymorphisms in cytokine genes may contribute to carcinogenesis. Interleukin (IL)-4 gene polymorphisms have been intensively studied with regard to their associations with cancer. However, the results of these previous studies remain inconclusive. The present study, therefore, aimed to conduct a meta-analysis of previously published studies in order to clarify the association of IL-4 with cancer risk. Eligible published articles were searched in Medline, PubMed, Embase and China National Knowledge Infrastructure databases up to March 2016. Odds ratios and 95% confidence intervals were used to identify potential associations between IL-4 genetic polymorphisms and the risk of cancer. A meta-analysis was then performed on 10,873 patients and 14,328 controls for IL-4 rs2243250 polymorphism, 3,970 patients and 5,686 controls for IL-4 rs2070874 polymorphism, and 1,896 patients and 2,526 controls for IL-4 rs79071878 polymorphism. A significant association with cancer risk was observed for rs2243250 and rs79071878 polymorphisms. In the subgroup analysis by cancer type, rs2243250 polymorphism was demonstrated to be associated with an increased risk of gastric cancer and breast cancer, rs2070874 polymorphism was correlated with leukemia and oral carcinoma, and rs79071878 polymorphism was relevant to bladder carcinoma risk. In the subgroup analysis by ethnicity, IL-4 rs2243250 polymorphism was demonstrated to be associated with cancer risk in both Caucasian and Asian populations, rs2070874 was associated with cancer risk in Asian populations, while rs79071878 polymorphism was associated with cancer risk in Caucasian populations. In conclusion, the present results suggested that the IL-4 rs2243250 and rs79071878 polymorphisms were associated with cancer susceptibility. Further subgroup analyses revealed that the effects of IL-4 gene polymorphisms on cancer risk may vary by cancer type and by ethnicity.

Influence of single nucleotide polymorphisms of cytokine genes on anti-HBs antibody production after hepatitis B vaccination in a Japanese young adult population.

Hepatitis B (HB) vaccination is one of the most efficient tools to prevent the transmission of the virus. Considerable variability exists in HB vaccine responses, with 5-10% of healthy Japanese adults demonstrating no response following a standard vaccination. Recently, polymorphisms of immune-regulatory genes, such as cytokine genes, have been reported to influence the immune response to HB vaccine. The aim of this study was to investigate the underlying mechanisms of the genetic association between several cytokine gene polymorphisms and the immune response to HB vaccination in a Japanese population. One hundred and twenty three vaccinated young adults were classified according to the level of antibody-titer (anti-HBs). Single nucleotide polymorphism typing for IFN-γ (+874, 3'-UTR), IL-10 (-591, -819, -1082), and TNF-α (-308, -857), was accomplished using the PCR-RFLP or SSP-PCR method. The TNF-α (-857) CC type and the IL-10 (-1082) AG type were present more frequently in the low titer group than in the high titer group. The TNF-α (-857) CC type was found to be significantly associated with low response of serum anti-HBs. The anti-HBs antibody was not readily produced in the IL-10 (-1082) AG and TNF-α (-857) CC haplotype. Conversely, the antibody was readily produced in the IL-10 (-1082) AA and TNF-α (-857) CC haplotype, and the IL-10 (-1082) AA and TNF-α (-857) CT haplotype, suggesting a high likelihood of the IL-10 (-1082) AG type to be included in the low anti-HBs group, and high anti-HBs antibody production in those with the TNF-α (-857) CT type. These SNPs may produce ethnically-specific differences in the immune response to HB vaccine in the Japanese population. J. Med. Invest. 63: 256-261, August, 2016.

Single nucleotide polymorphisms for genes encoding cytokines in the context of cardiac surgery. Part I: Heart transplantation.

Cardiovascular diseases remain the leading cause of death in Poland and other countries of the European Union. Patients with end-stage heart failure constitute a patient subgroup for whom the treatment of choice is heart transplantation. Despite advances in immunosuppressive therapy, acute or chronic graft rejection occurs in 20-30% of cases in the first six months after transplantation. The significance of the immune response and inflammation in graft rejection implies the important role of cytokines. Molecular markers are sought to facilitate risk assessment and improve patient care. At present, genetic tests are not used for this purpose, but studies aiming to rectify that have been conducted for years, including studies on single nucleotide polymorphisms of cytokine genes. This paper presents the results of research on the single nucleotide polymorphisms (SNPs) of the IL-2, IL-4, IL-6, IL-10, TGF-β1, PDGF, VEGF, and TNF-α genes in conjunction with heart transplantation. The analyzed data do not allow for reliable application of these genetic tests in clinical practice, but suggest that it is a promising direction which may improve the options of treatment individualization in the future.

The role of single nucleotide polymorphisms of cytokine genes in viral infections

Gene polymorphisms result from evolutionary processes representing mutations that survive in the population with a frequency higher than 1%. The most investigated type of gene polymorphisms are single nucleotide polymorphisms (SNPs). The SNPs of IL-12B (rs 3212227) A/C among a population of kidney graft CMV-seropositive recipients have an impact on a clinical events in cytomegalovirus (CMV) disease. Constitutive -308 G/A TNF-? polymorphism (rs1800629) is related to the susceptibility of HR-HPV-associated cervical dysplasia and cancer. SNP located 3 kb upstream of the IL- 28B gene (rs12979860) seems to be the strongest host genetic predictor of sustained virologic response (SVR) in hepatitis C genotype 1 patients. It is very important to identify viral and host genetic markers that may facilitate the risk of developing viral disease or some viral-associated cancers. In addition, these markers could be useful in the choice of effective treatments and preventive strategies against virally induced infection.

Role of Tumor Necrosis Factor-Alpha (C-863A) Polymorphism in Pathogenesis of Inflammatory Bowel Disease in Northern India

Inflammatory bowel diseases (IBDs), namely ulcerative colitis (UC) and Crohn's disease (CD), are characterized by chronic and idiopathic inflammatory conditions of gastrointestinal tract that are immunologically mediated. Single nucleotide polymorphisms in cytokine genes have been reported to modulate inflammation. Therefore, we analyzed the association of pro/anti-inflammatory cytokine genes polymorphism with IBD susceptibility. Genotyping of interleukin (IL)-4 repeat polymorphism in intron-3, IL-10 (G-1082A and C-819T), and tumor necrosis factor-alpha (TNF-A) (-1031 T>C, -863 C>A, and -857 C>T) was performed in 153 patients with IBD and in 207 controls. TNF-A -863 AA genotype was associated with enhanced IBD susceptibility (odds ratio (OR), 4.82; 95% confidence interval (CI), 2.60-8.96), more so for UC (OR, 5.79; 95% CI, 2.99-11.21), Crohn's disease [CD] (OR, 3.13; 95% CI, 1.16-8.47). TNF-A T/C/T (OR, 4.40; 95% CI, 1.64-11.81) and C/A/C (OR, 4.15; 95% CI, 2.48-6.96) haplotypes were associated with increased IBD risk. The frequency of IL-4, B2 carrier (B1/B2 + B2/B2) was significantly lower in left-sided UC (17.1%) than proctosigmoiditis (47.6%); p, 0.016. In contrast, TNF-A -863 AA genotype frequency was much higher in pancolitis (45.5) than in proctosigmoiditis (14.2); p, 0.037. Variant genotypes of IL-4 (B1/B2 + B2/B2) were absent in colonic type CD. IL-10 polymorphisms did not demonstrate any association with IBD. None of the polymorphisms were associated with steroid treatment and surgery. The present study depicts that high-producing genotype of TNF-A (-863 AA) was associated with increased risk of IBD more so with UC. Similarly, combined effect of TNF-A polymorphisms in haplotype analysis demonstrated additively increased risk of IBD.

M13.1 Association of single nucleotide polymorphisms in cytokine genes with preeclampsia in a Sinhalese population from Sri Lanka

Prabha Andraweera1, Miss Rachael Nowak1, Steven Thompson1, Vajira Dissanayake2, Rohan Jayasekara2, Gus Dekker3, Claire Roberts1. 1Discipline of Obstetrics and Gynaecology, Research Centre for Reproductive Health, University of Adelaide, Australia; 2Human Genetics Unit, Faculty of Medicine, University of Colombo, Sri-Lanka; 3Women’s and Children’s Division, Lyell McEwin Hospital, Elizabeth Vale, South Australia

Single Nucleotide Polymorphisms of Cytokine Genes are Associated with Fibrosis of the Intrahepatic Bile Duct Wall in Human Clonorchiasis

This study examined the association of cytokine gene polymorphisms with intrahepatic bile duct wall fibrosis in human clonorchiasis. A total of 240 residents in Heilongjiang, China underwent ultrasonography, blood sampling, and stool examination. Single nucleotide polymorphism (SNP) sites for IFN-gamma (+874 T/A), IL-10 (-1,082 G/A, -819 C/T, -592 C/A), TNF-alpha (-308 G/A), and TGF-beta1 (codon 10 T/C, codon 25 G/C) genes were observed with the TaqMan allelic discrimination assay. No significant correlation was observed between individual cytokine gene polymorphisms and intrahepatic duct dilatation (IHDD). Among individuals with clonorchiasis of moderate intensity, the incidence of IHDD was high in those with IFN-gamma intermediate-producing genotype, +874AT (80.0%, P = 0.177), and in those with TNF-alpha low-producing genotype, -308GG (63.0%, P = 0.148). According to the combination of IFN-gamma and TNF-alpha genotypes, the risks for IHDD could be stratified into high (intermediate-producing IFN-gamma and low producing TNF-alpha), moderate, and low (low-producing IFN-gamma and high producing TNF-alpha) risk groups. The incidence of IHDD was significantly different among these groups (P = 0.022): 88.9% (odds ratio, OR = 24.0) in high, 56.5% (OR = 3.9) in moderate, and 25.0% (OR = 1) in low risk groups. SNP of IFN-gamma and TNF-alpha genes may contribute to the modulation of fibrosis in the intrahepatic bile duct wall in clonorchiasis patients.

Zytokine in der Diagnostik der Uveitis

Non-infectious uveitis is characterised by a breakdown of immunological tolerance. Several cytokines could be identified as important regulators of the immune system during induction, progression and remission of ocular inflammation in animal models. This review analyses the current knowledge on cytokines in uveitis and their relevance for diagnostic purposes. Elevated levels of pro-inflammatory and decreased concentrations of anti-inflammatory cytokines were detected in the aqueous humor and serum of uveitis patients. In addition, single nucleotide polymorphisms of cytokine genes altering expression levels were found to be associated with an increased risk of uveitis. These studies, however, included only small or heterogeneous patient populations. Moreover, the reported data are partially inconsistent. Thus, the analysis of cytokines does not play a role in the routine diagnostic process of uveitis so far. New multiplex techniques for cytokine analysis will provide comprehensive data for a better understanding of the complex regulation of the immune system during intraocular inflammation. This may lead to a quantitative description of the immunological mechanisms of uveitis in the individual patient to improve the prognostic evaluation and select the most efficient treatment.

Single nucleotide polymorphisms of cytokine genes in the healthy Slovak population

Cytokines are molecules that control and modulate the activities of numerous target cells via binding to specific receptors. The observed differences in the cytokine production among individuals can be, at least partially, explained by gene polymorphisms. Several cytokine gene polymorphisms have been identified to play a role in susceptibility to various diseases, including autoimmune, infectious, allergic or cardiovascular diseases. The aim of the current study was to determine allele and genotype frequencies of 22 polymorphisms in 13 cytokine genes in the healthy Slovak population and to compare them with data available from six populations from Central and Southern Europe. A polymerase chain reaction with sequence-specific primers was used to genotype polymorphisms within genes encoding IL-1alpha, IL-1beta, IL-1R, IL-1RA, IL-4Ralpha, IL-12, IFN-gamma, TGF-beta, TNF-alpha, IL-2, IL-4, IL-6 and IL-10 in a sample of 140 unrelated Slovak subjects. The allelic distribution of all polymorphisms in the Slovak population was very close to that in the geographically and historically closest populations in Central Europe--the Czech and the Polish. However, several differences were found between the Slovak and four populations from Southern Europe. The obtained data represent a basis for further studies on association of cytokine gene polymorphisms with some diseases.

Role of Single Nucleotide Polymorphisms of Cytokine Genes in Spontaneous Preterm Delivery

Intra-amniotic infections are implicated in spontaneous preterm delivery (PTD). Certain genetic polymorphisms are associated with increased production of proinflammatory and/or decreased production of anti-inflammatory cytokines, thereby possibly promoting PTD. We determined the relationship between maternal and fetal cytokine gene polymorphisms with occurrence and severity of spontaneous PTD (PTD after spontaneous-onset preterm labor and/or preterm prelabor rupture of membranes) and their association with intrauterine inflammation and infection. DNA from buccal brushings of 80 preterm (gestation < 35 weeks) and 80 matched term mother-infant pairs was assayed for tumor necrosis factor α (TNF-α [−308G/A]), interferon-γ (IFN-γ [+874A/T]), interleukin-6 (IL-6 [−174C/G]), interleukin-10 (IL-10 [−1082G/A, −819C/T, −592C/A]), and transforming growth factor β 1 (TGF-β 1 [T/Ccodon10,G/Ccodon25]) by using polymerase chain reaction (PCR) with sequence-specific primers. The presence of histologic chorioamnionitis was determined for PTDs. Conditioned on maternal IFN-γ genotypes, fetal high IFN-γ producing allele (IFN-γ[+874T]) was associated with spontaneous PTD (odds ratio = 2.3 [1.2–4.4]). Among preterm deliveries, maternal low TGF-β 1 (TGF-β 1 [codon10C]) producing genotypes correlated negatively with gestation. Fetal TNF-α (−308G) was significantly associated with histologic chorioamnionitis. Underlying genitourinary infections and/or inflammation were significantly associated with maternal and fetal IL-6 (−174G), fetal TNF-α (−308GG), and fetal IL-10 (−1082A). We conclude that certain fetal and maternal cytokine gene polymorphisms may be associated with occurrence and/or severity of spontaneous PTD and with intrauterine inflammation and infection.

Cytokine gene polymorphisms involved in chronicity and complications of anterior uveitis

The aim of this study was to identify key cytokine polymorphisms associated with disease susceptibility, clinical phenotype, and outcome in patients with chronic anterior uveitis (CAU) as compared to those with recurrent self-limiting anterior uveitis (RAU). One hundred fifty seven British Caucasian patients with anterior uveitis were identified and divided into those where the inflammatory process lasted less than 3 months (RAU = 118) and those where the inflammation persisted longer than 3 months (CAU = 39). Patients with CAU were further sub-divided into idiopathic CAU, CAU associated with systemic disease, CAU with and without complications (posterior synechiae, posterior subcapsular lens opacity, raised intraocular pressure, cystoid macular oedema, and poor response to treatment). Sixty-six healthy controls were ethnically matched. TaqMan PCR amplification was used to genotype five single nucleotide polymorphisms in cytokine genes; IL-1RN+2018, IL-6−174, IL-10−1082, TNF−238, TNF−308 and these were correlated with clinical phenotype.

Genotyping of single nucleotide polymorphisms in cytokine genes using real-time PCR allelic discrimination technology

Single nucleotide polymorphisms (SNPs), particularly those within regulatory regions of genes that code for cytokines often impact expression levels and can be disease modifiers. Investigating associations between cytokine genotype and disease outcome provides valuable insight into disease etiology and potential therapeutic intervention. Traditionally, genotyping for cytokine SNPs has been conducted using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), a low throughput technique not amenable for use in large-scale cytokine SNP association studies. Recently, Taqman ® real-time PCR chemistry has been adapted for use in allelic discrimination assays. The present study validated the accuracy and utility of real-time PCR technology for a number of commonly studied cytokine polymorphisms known to influence chronic inflammatory diseases. We show that this technique is amenable to high-throughput genotyping and overcomes many of the problematic features associated with PCR-RFLP including post-PCR manipulation, non-standardized assay conditions, manual allelic identification and false allelic identification due to incomplete enzyme digestion. The real-time PCR assays are highly accurate with an error rate in the present study of <1% and concordance rate with PCR-RFLP genotyping of 99.4%. The public databases of cytokine polymorphisms and validated genotyping assays highlighted in the present study will greatly benefit this important field of research.

TNF-α, IL-8 and CD14 gene polymorphisms in Helicobacter pylori-positive duodenal ulcer patients

Background: The activation of inflammatory cytokines plays an important role in the pathogenesis of the development of Helicobacter pylori (HP)-positive duodenal ulcer (DU). Cytokine production is determined in part by the nucleotide polymorphisms of cytokine genes. The aim was to evaluate the frequency of TNF-α, IL-8 and CD14 gene polymorphisms in HP-positive DU patients. Patients. 69 patients with HP-positive DU (mean age 46.8 years) and 47 HP- positive healthy blood donors (mean age 42.6 years) were included in the study. Methods: The TNFα-308A (G to A) gene polymorphism was determined by NcoI RFLP (restriction fragment length polymorphism), the IL-8–251A (T to A) gene polymorphisms by ARMS (amplification refractory mutation system) methods and the CD14–159T (C to T) polymorphism by melting point analysis.