Promoter Region Of Interleukin-10 Gene Research Articles

The Involvement of Interleukin-10 Promoter Genetic Polymorphism in Epstein-Barr Virus-Associated Nasopharyngeal Carcinoma from North Africa

Objectives: Interleukin-10 (IL10) is a pro-inflammatory cytokine that plays a pivotal role in inflammatory diseases as well as in the pathogenesis of diverse tumors, including nasopharyngeal carcinoma (NPC). The present study was designed to evaluate the importance of the functional promoter polymorphisms of IL10 (-1082 A>G and -592 A>C) in the development of NPC. Methods: A total of 384 patients with NPC were recruited into this study, together with 375 matched control subjects. Polymorphisms within the promoter region of IL10 gene were analysed using PCR-RFLP method. Results: We report a lack of association between IL10 polymorphisms and NPC in the overall population (P>0.05). However, the 1082 A>G polymorphism was significantly associated with the susceptibility to NPC among young patients (age ?30 years). The GG genotype was found to be associated with a significantly higher risk of NPC as compared with the AA genotype among young patients (OR=2.534; 95% CI, 1.189–5.398, P=0.016). Conclusion: The results of the present study indicate an association between IL10 -1082 GG genotype and NPC among young North African subjects. This difference in IL10 polymorphism association with different ages at the onset of NPC suggests that the younger and older onset patients are genetically different and may involve different mechanisms. Keywords: Nasopharyngeal Carcinoma, IL10, North African population

Associations of Single Nucleotide Polymorphisms in IL-18 Gene with Plasmodium falciparum-Associated Malaria

IntroductionInterleukin-18 (IL-18) is a pro-inflammatory cytokine, reported to be involved in the initial immune responses against malaria. Genetic variations in the host are an important factor that influences the etiology of malaria at several disease levels. Polymorphisms within the IL-18 gene are associated with susceptibility and clinical outcome of several diseases.MethodsWe genotyped single nucleotide polymorphisms (SNPs) in IL-18 of patients infected with Plasmodium falciparum with varying extent of parasitemia and different age groups.ResultsSNPs rs5744292 (OR = 70.446; 95% CI = 4.318–1149.323; p<0.0001) and rs544354 (OR = 1.498; 95% CI = 1.088–2.063; p=0.013) were found to be significantly associated with parasitemia in P. falciparum-infected patients when compared with healthy control subjects. SNP rs5744292 (OR = 7.597; 95% CI=1.028–56.156; p=0.019) was associated with increased parasite density in infected patients. SNPs rs544354 (OR 0.407; 95% CI=0.204–0.812; p = 0.009) and rs360714 (OR of 0.256; 95% CI=0.119–0.554; p = 0.001) were significantly associated with parasite density in an age-dependent manner, with the risk alleles present more frequently among the younger (1–9 years) patients. Several haplotypes were found to have a significant association with parasitemia. In-vitro expression analysis using luciferase reporter assay showed that SNPs rs1946518 and rs187238 in the IL-18 gene promoter region and rs360728 and rs5744292 in the 3ʹ-untranslated region of the IL-18 gene were associated with enhanced transcriptional activity.ConclusionOur results suggest that polymorphisms within the IL-18 gene are associated with the susceptibility to P. falciparum infection and related parasitemia among groups with different parasite density and across various age groups.

Evaluation of influence of IL-6 C-572G gene polymorphism and clinical factors on positive platelet antibody test

Background: Interleukin-6 (IL-6) promotes antibody production. The objective of this study was to investigate whether IL-6 C-572G single nucleotide polymorphisms (SNP) and clinical factors are associated with positive platelet antibody test. Materials and methods: Thirty platelet recipients with platelet antibodies (responders) and 20 platelet recipients without platelet antibodies (non-responders) were randomly selected. The -572 C&gt;G (rs 1800796) SNPs in the promoter region of IL-6 gene were genotyped by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method. Solid phase red cell adherence assay (SPRCA) was used for platelet antibody detection. Results: Age, sex, percentage patients with benign diseases, and percentage of patients with homozygotes for the C allele at position -572 of the IL-6 gene were similar between responders and non-responders. Although the amounts of platelets pheresis transfused to patients with hematologic diseases were higher than those of non-hematologic diseases (47.2 ± 54.2 vs. 17.4 ± 13.8 units, p = 0.019), detection rate of platelet antibodies was lower in patients with hematologic diseases than that in patients with non-hematologic diseases (42.3% vs. 79.2%, p = 0.01). Conclusion: There was no association between IL-6 C-572G gene polymorphism and positive reactivity in solid phase platelet antibody detection method in platelet recipients.

IL-10 gene promoter region polymorphisms and their association with celiac disease.

in celiac disease (CD), immune response activation results in local cytokine network impairment. Interleukin 10 (IL-10) is a key anti-inflammatory cytokine in the prevention of inflammatory conditions. to analyze the association of single nucleotide polymorphisms in the IL-10 gene promoter region with CD in a population of Misiones Province, Argentina. DNA from whole blood was extracted from 40 patients with CD and 80 controls and the IL-10 gene promoter region containing polymorphisms rs1800896A/G, rs1800871T/C and rs1800872A/C was amplified. Risk was established by calculating odds ratios (OR) and statistical significance was considered as p < 0.05. there were no significant differences in rs1800896 genotype distribution between celiac patients and controls. The frequency of the CC genotype for rs1800871T/C and rs1800872A/C was lower among celiac patients (35 % vs 65 %; p = 0.002). CD risk was associated with carriers of the more uncommon T allele of rs1800871T/C and the more uncommon A allele of rs1800872A/C, with a dominant model (OR = 2.79; 95 % CI: 1.27-6.09; p = 0.01). A risk effect was found for haplotype ATA (OR = 3.05; 95 % CI: 1.25-7.46; p = 0.01). carriers of the less common T allele of rs1800871T/C and the less common A allele of rs1800872A/C in the IL-10 gene promoter are at high risk of CD with a dominant model. There was no risk for rs1800896A/G. The ATA haplotype showed an association with CD development.

English

The promoter region of human interleukin-10 (IL-10) gene is highly polymorphic while the first intronic region of interferon gamma (IFN-) gene is also highly polymorphic. These polymorphisms are associated with susceptibility or resistance to Schistosoma haematobium infection. Schistosomiasis is known to be a highly inflammatory disease that requires the delicate balance of pro- and anti-inflammatory cytokines. The polymorphisms are associated with low, moderate or high cytokine production resulting in exacerbation of the infection leading to pathological severity. Urine filtration technique was used for diagnosis the S. haematobium. Whole blood samples were collected from 400 children aged between 6 to 13 years. Molecular determination of polymorphism related to resistance or susceptibility to infection was performed using the allele-specific polymerase chain reaction. SNPs in the IL-10 and IFN- cytokine genes were examined in blood samples from 400 school-aged children. Schistosomiasis was detected in 49.8% (199). For IFN- +874A/T, the distribution of TT, TA and AA was 7, 41 and 51% respectively. An analysis of the polymorphisms on IL-10 -1082G/A showed that most of the samples were heterozygous (47% GA) whereas AA (32%) and GG (21%) were monozygous. SNPs within the promoter region of IL-10 gene and in the intronic region of IFN- have been associated with altered profiles of circulating IL-10 and IFN-. Our findings suggest that IL-10 and IFN- polymorphisms participate in the progression of schistosomiasis rather than in its initial development in school aged children. It is recommended to study more polymorphisms in the other cytokines implicated in schistosomiasis. Key words: Polymorphism, cytokine, Schistosoma haematobium, interleukin-10, interferon gamma. &nbsp; &nbsp

Association of interleukin-6 rs1800796 polymorphism with reduced cognitive performance in healthy older adults

With increasing life expectancy, age-associated cognitive impairment is an escalating problem worldwide. Inflammation is one of the features that characterises cognitive decline and can stimulate neurodegeneration. Interleukin 6 (IL-6) is a cytokine frequently associated with a pro-inflammatory phenotype and increased levels have been associated with the pathogenesis of dementia. The rs1800796 polymorphism in the promoter region of IL-6 gene was previously shown to influence IL-6 expression and therefore we hypothesised this gene polymorphism would be associated with IL-6 plasma levels and cognitive performance of older adults.The present study investigated the association of the rs1800796 polymorphism on plasma IL-6 levels and cognition in healthy older adults (n = 207, 74.6 ± 3.4 years, 51% female) that participated in a Pan-European project (MyoAge). The participants were assessed for working memory capacity, executive functioning, episodic memory and global cognition using the Cambridge Neuropsychological Test Automated Battery CANTAB. Fasting plasma IL-6 levels were measured by ELISA and genotyping was performed using the KASP assay.Results showed that the rs1800796 polymorphism was in Hardy-Weinberg equilibrium (P = .16) with the minor allele (C) showing a frequency of 6.3%. There were no differences in plasma IL-6 concentrations between the GG-homozygotes and C-allele carriers (P = .22). The C-allele carriers performed worse on a measure of executive functioning (P = .035) and had lower global cognitive scores (P = .045), compared to GG-homozygotes. These differences remained significant after accounting for age, sex and prior cognitive abilities (P < .05 for both). There were no differences in measures of memory (episodic and working) between the genotypes group.These findings suggest that the rs1800796 variant may be detrimental for executive functioning, but not memory, in healthy older adults.

Analysis of interleukin-10 gene polymorphisms in patients with chronic periodontitis and healthy controls.

Interleukin-10 (IL-10) is an anti-inflammatory cytokine that has important roles in the periodontal diseases. The IL10-1082, -819, and -592 polymorphisms in the promoter region of IL-10 gene have been associated with various IL-10 expressions. The aim of this study was to investigate the association between these gene polymorphisms with chronic periodontitis in a sample of Iranian populations from Southeast of Iran. IL-10 single nucleotide polymorphisms were analyzed in 210 patients with chronic periodontitis (CP) and 100 individuals without CP by polymerase chain reaction-restriction fragment length polymorphism method. Statistical analysis of data was performed using the Chi-square test. The risk associated with single alleles, genotypes, and haplotypes were calculated by performing a multiple logistic regression analysis to estimate the odds ratio (OR) and 95% confidence interval (CI). P < 0.05 for statistical significance. The prevalences of AG and GG genotypes of IL10-1082 were significantly different between CP and control groups in comparison to AA genotype (OR = 2.671; CI = 1.482-4.815; P = 0.001 for AG vs. AA, OR = 4.151; CI = 2.128-8.097; P < 0.001 for GG vs. AA). In addition, subjects with at least one IL10-1082-G allele were significantly had an increased risk for CP (OR = 2.157; CI = 1.531-3.038; P < 0.001). The distribution of the IL10-819 and IL10-592 genotypes was not different between CP and control subjects (P = 0.109 and P = 0.139, respectively). The combination of different genotypes showed that GCC haplotype was significantly different between groups (OR = 4.379; CI = 1.077-17.807; P = 0.039). The results demonstrated that IL10-1082 polymorphism was a putative risk factor for chronic periodontitis and associated with increased susceptibility to CP.

Genetic variations of IL-10: Identification of novel variations and evaluation of the impact of the SNPs/haplotype in the promoter region with the progression of Oral Squamous Cell Carcinoma in Indian population

The correlation of interleukin 10 (IL-10) with the outbreak and progression of cancer has been well established as it contributes to tumor immune evasion. Convincing number of evidences has been accumulated to reflect the critical correlation between IL-10 polymorphism and tumorogenesis. Several polymorphic sites at promoter regions have been reported to be associated with cancer susceptibility. The purpose of this study was to examine the effect of modulated genotypes in the promoter region of IL-10 gene with life-style habits in oral squamous cell carcinoma (OSCC) in the Indian population. A total of 300 subjects (100 OSCC, 50 precancer and 150 healthy controls) were recruited in this study. The IL-10 promoter region was amplified in 14 overlapping fragments by PCR and further screened through the high throughput technique of denaturing high-performance liquid chromatography (dHPLC) followed by sequencing. We identified three novel variations at positions (−924, −1045 & −1066); we also found some known SNPs (−592C/A, −657G/A, −851G/A, −819C/T, −1082A/G). The identified novel variations were submitted to the NCBI Gene Bank (accession numbers KT153594, KT291742 and KT291743). We also noticed a significant association of polymorphisms (−592C/A, −819C/T and −1082A/G) individually as well as in combination (haplotypes) along with lifestyle habits for the risk of oral carcinoma (p<0.0001). We have reported three novel SNPs in the Indian population for the first time, and these SNPs may be associated with OSCC. Besides, we showed the first evidence of IL-10 haplotypes, i.e., CCG and CTG, may act as a biomarker for early detection of oral pre-cancerous/cancerous lesions or treatment management of oral carcinoma.

Effect of interleukin-10 gene promoter polymorphisms -1082 G/A and -592 C/A on response to therapy in children and adolescents with chronic hepatitis C virus infection

Background and aimStudying predictors of response to therapy for hepatitis C virus (HCV) infection in children may help avoid the inappropriate use of currently available costly therapy associated with numerous adverse effects. We tested the hypothesis that inheritance of single nucleotide polymorphisms (SNPs) of the interleukin-10 (IL-10) promoter gene might influence response to HCV treatment. Patients and methodsThe impact of SNPs, -1082 G/A and -592 C/A, in the promoter region of IL-10 gene, on response to HCV therapy was assessed in a cohort of 40 children treated with a combination of pegylated interferon (Peg-IFN) α2b and ribavirin. ResultsSustained virological response was achieved in 48.7%. High viral load was associated with non-response to therapy. There was no association between histopathological degree of inflammation or fibrosis and response to therapy. There was no direct statistically significant association between polymorphisms in the IL-10 gene (-1082G/A and -592 C/A) as regards inflammation or response to therapy in children. As for the SNP -592 C/A; there was a statistically significant association with the score of fibrosis (P<0.004), concluding that the A allele was protective from moderate and severe fibrosis. Meanwhile the SNP -1082G/A did not show any association with the fibrosis score. ConclusionWe could not associate response to therapy for HCV with IL-10 polymorphisms -1082 G/A and -592 C/A. For the SNP -592 C/A, the A allele protected from moderate and severe fibrosis.

Lack of association between rs1800795 (-174 G/C) polymorphism in the promoter region of interleukin-6 gene and susceptibility to type 2 diabetes in Isfahan population.

Background:Type 2 diabetes mellitus (T2DM) is an inflammatory autoimmune disease that mostly affects older adults. The etiology of T2DM includes both genetic and environmental factors. rs1800795 (−174 G/C) single nucleotide polymorphism (SNP) linked with autoimmune disorders predispositions, identified by Genome-Wide Association Study among genes, which immunologically related is considerably over signified. The goal of this study was to evaluate the association between rs1800795 (−174 G/C) polymorphisms in the promoter of interleukin-6 (IL-6) gene with susceptibility to T2DM in a subset of the Iranian population.Materials and Methods:In this case–control study, 120 healthy subjects and 120 patients with T2DM were included. Genomic DNA obtained from whole blood samples and the polymerase chain reaction was used to amplify the fragment of interest contain rs1800795 SNP, restriction fragment length polymorphism method was applied for genotyping of the DNA samples with NlaIII as a restriction enzyme. SPSS for Windows software (version 18.0, SPSS, Chicago, IL, USA) was performed for statistical analysis.Results:No significant differences were found between healthy controls and T2DM patients with respect to the frequency distribution of the cytokine gene polymorphism investigated. Odds ratio, adjusted for sex, age, and smoking status has displayed similar outcomes.Conclusion:These results indicated that the rs1800795 SNP is not a susceptibility gene variant for the development of T2DM in the Isfahan population. Further studies using new data on complex transcriptional interactions between IL-6 polymorphic sites are necessary to determine IL-6 haplotype influence on susceptibility to T2DM.

Interleukin 6 promoter polymorphisms in normal Han Chinese population: frequencies and effects on inflammatory markers.

Interleukin 6 (IL-6) is a cytokine involved in different physiologic and pathophysiologic processes, and circulating levels of IL-6 differ greatly between individuals, but the results have not always been concordant among diverse populations. The aim of present study was to determine the prevalence of the 3 polymorphisms (-174G/C, -597G/A, and -634C/G) in the IL-6 gene promoter region and their effects on inflammatory markers in normal Han Chinese population. A total of 232 subjects (143 men and 89 women; mean age, 51.37 ± 17.63 years; range, 22-88 years) of unrelated healthy Han Chinese in Jinangsu area (south of China) were enrolled. Genotyping of the 3 polymorphisms were performed by polymerase chain reaction and restriction fragment length polymorphism combined with gel electrophoresis and then confirmed by direct sequencing. Among the 232 individuals studied, 231 carried the GG wild type of -174G/C; only 1 carried the GC genotype. For -597G/A polymorphism, individuals all carried the GG wild type; the GA or AA genotypes were not detected. The frequencies of -634C/G genotypes CC, CG, and GG were found to be 59.48%, 37.07%, and 3.45%, respectively, the derived allele frequencies for the C and G alleles were 78.02% and 21.98%. There were no significant differences in age, sex, body mass index, or lipids parameters between the -634 CC and CG+GG genotypes. However, individuals with CC genotype showed lower levels of high-sensitivity C-reactive protein and IL-6 than those with CG+GG genotype. IL-6 -174G/C and -597G/A are rare, but -634C/G is common in Han Chinese population, and the -634G allele is associated with circulating levels of IL-6 and C-reactive protein.

Role of Interleukin-10 Gene Promoter Region Polymorphism in the Development of Chronic Lymphoid Leukemia.

Relationship between interleukin-10 (IL-10) gene G-1082A (rs1800896) polymorphism and the risk of development and stages of chronic lymphoid leukemia is studied in ethnic Russian residents of the Kirov region of Russia. Associations of allele -1082A and genotypes (-1082AA/-1082AG) with the risk of chronic lymphoid leukemia are detected (OR=1.39, 95%CI=1.09-1.78 and OR=1.66, 95%CI=1.09-2.54, respectively). In addition, association of 1082AA genotype with late stages of the disease by the moment of diagnosis is detected. These data indicate that IL-10 polymorphism G-1082A may be involved in the pathogenesis of chronic lymphoid leukemia.

Interleukin-10 gene promoter region polymorphisms are not associated with BCG osteitis in vaccinated infants.

Complications arising from bacille Calmette-Guérin (BCG) vaccination were recorded in a national register in Finland until 1988. In the period 1960-1988, 222 patients suffered from BCG osteitis. To evaluate whether single nucleotide polymorphisms (SNPs) in the promoter region of the gene encoding interleukin 10 (IL-10) are associated with BCG osteitis after vaccination in neonates. Blood samples of 132 former BCG osteitis patients now aged 21-49 years were analysed in a controlled study for IL10 rs1800896 (-1082G/A), rs1800871 (-819C/T), rs1800872 (-592C/A) and rs1800890 (-3575T/A) polymorphisms. The frequencies of genotypes of IL10 rs1800896, rs1800871, rs1800872 and rs1800890, the frequencies of variant genotypes and the frequencies of major or minor alleles did not differ between patients and controls. Furthermore, the frequencies of the eight possible combinations of the three IL10 alleles located close to each other (IL10 rs1800896, IL10 rs1800871 and IL10 rs1800872) were surprisingly similar. Our results suggest that polymorphisms of the IL-10 encoding gene do not play a central role in the development of complications due to BCG vaccination, although the IL10 gene, especially IL10 rs1800896 (-1082G/A) polymorphism, is known to be associated with tuberculosis risk in Europeans and North Americans.

Gene polymorphisms of interleukin-10 and transforming growth factor beta in allergic rhinitis

BackgroundAllergic rhinitis (AR) is a polygenic inflammatory disorder of the upper respiratory airway with an increasing prevalence worldwide. Interleukin-10 (IL-10) and transforming growth factor-beta (TGF-β), as two cytokines with pleiotropic effects on both innate and adaptive immunity, play important roles in allergic responses. Therefore, this study was performed to evaluate the associations of five polymorphisms of IL-10 and TGF-β genes with AR. Materials and methodsNinety-eight patients with AR along with 140 healthy volunteers with no history of AR and with the same ethnicity of the patients were recruited in this study. Genotyping was done for three polymorphisms in promoter region of IL-10 gene (rs1800896, rs1800871, rs1800872), and two polymorphisms in the exonic region of TGF-β1 gene (rs1982037, rs1800471) using PCR sequence-specific-primers method. ResultsA allele and AA genotype in rs1800896 of IL-10 and TT genotype in rs1982037 in TGF-β were significantly less frequent in the patients than in controls. While the C allele and the CG genotype in rs1800471 in TGF-β1 were associated with a higher susceptibility to AR. C/C and T/C haplotypes (rs1982037, rs1800471) in TGF-β1 gene and A/C/A, A/T/C and G/C/A haplotypes (rs1800896, rs1800871, rs1800872) in IL-10 gene were found with higher frequencies in patients than controls. Patients with CC genotype in rs1800871 in Il-10 had significantly lower levels of IgE. ConclusionWe found that certain genetic variants in IL-10 and TGF-β polymorphisms were associated with susceptibility to AR as well as some clinical parameters in the patients with AR.

A functional polymorphism in the promoter region of interleukin-10 gene increases the risk for spontaneous abortions--a triad study.

Spontaneous abortion or miscarriage is the natural death of an embryo or foetus in the early stages of prenatal development. Interleukin-10 is an anti-inflammatory cytokine, produced by human cytotrophoblasts, and defects in its production result in specific pathological conditions during pregnancy. The present study is aimed to evaluate the association of IL-10 -1082G/A polymorphism in spontaneous abortions by comparing foetal, maternal and paternal groups--a triad study. A total of 50 families with spontaneous abortions and 60 families with medically terminated pregnancies were considered for the present study. DNA from foetal tissue and parental blood samples were extracted, and the genotype analysis of IL-10 -1082G/A promoter polymorphism was carried out by amplification refractory mutation system-polymerase chain reaction followed by agarose gel electrophoresis. A statistical analysis was applied to test for the significance of the results. There was a statistically significant difference in the distribution of AA genotypes and A allele of IL-10 -1082G/A between the two family groups among foetuses (P = 0.0002) and mothers (P = 0.00005). The paternal group showed no significant difference in the genotype distribution of IL-10 between cases and controls. In conclusion, IL-10 G-1082A gene promoter polymorphism may act as a major genetic regulator in the etiology of spontaneous abortions.

Ethnicity-stratified analysis of the association between IL-18 polymorphisms and systemic lupus erythematosus in a European population: a meta-analysis.

We performed a meta-analysis to identify the association between polymorphisms in the promoter of interleukin-18 (IL-18) and susceptibility for systemic lupus erythematosus (SLE) . Genotype data for three single-nucleotide polymorphisms (SNPs rs360719, rs1946518, and rs187238) in the IL-18 promoter were extracted from 20 studies of three different ethnicities (European, Asian, and South American). Data from each ethnicity group and their combinations were analyzed. We found distinct evidence of an association between rs360719 and SLE (P = 0.001) in the European/South American group [odds ratio (OR) 1.31 per C allele, 95% confidence interval (CI) 1.11-1.53]. Stratification analysis by ethnicity showed a significant association between rs360719 and SLE in the European population (OR 1.33 per C allele, 95% CI 1.11-1.61, P = 0.003) and a lesser effect in the same direction in the South American population (OR 1.18). A significant association was also identified between rs1946518 and SLE in the European population (OR 1.16 per A allele, 95% CI 1.03-1.30, P = 0.017), although there was no association in the Asian or the combined European/Asian population. We also examined genome-wide association study (GWAS) data from an Asian subpopulation (Chinese) for the association between rs1946518 and SLE, but found no association (P = 0.83). The third SNP, rs187238, was not significantly associated with SLE in any of the populations examined. In summary, this study identified a significant association between SLE and two SNPs within the IL-18 gene promoter region (rs360719 and rs1946518) in a European population, but not in populations of Asian origin.

Transcription factor myocyte enhancer factor 2D regulates interleukin-10 production in microglia to protect neuronal cells from inflammation-induced death.

BackgroundNeuroinflammatory responses have been recognized as an important aspect in the pathogenesis of Parkinson’s disease (PD). Transcriptional regulation plays a critical role in the process of inflammation. Transcription factor myocyte enhancer factor 2D (MEF2D) is identified as a central factor in transmission of extracellular signals and activation of the genetic programs in response to a wide range of stimuli in several cell types, including neurons. But its presence and function in microglia have not been reported. We therefore investigated the effect of MEF2D in activated microglia on the progress of neuroinflammation and the survival of neurons.MethodsBV2 cells and primary cultured glial cells were stimulated with lipopolysaccharide (LPS). Samples from cells were examined for MEF2D expression, interleukin-10 (IL-10), and tumor necrosis factor alpha (TNF-α) by immunoblotting, quantitative real-time PCR (qPCR) or enzyme-linked immunosorbent assay (ELISA). The activity of MEF2D was examined by electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP). Recombinant lentivirus expressing shRNA specific to MEF2D was used to silence MEF2D expression in BV2 cells. The role of IL-10 transcriptionally induced by MEF2D on neuronal survival was assessed by anti-IL-10 neutralizing antibody. The survival of neurons was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. Male C57bl/6 mice were used to establish an acute PD model. Brain sections and cell slides were tested by immunofluorescence.ResultsWe demonstrated that MEF2D was present in microglia. Activation of microglia was associated with an increase in MEF2D level and activity in response to different stimuli in vivo and in vitro. MEF2D bound to a MEF2 consensus site in the promoter region of IL-10 gene and stimulated IL-10 transcription. Silencing MEF2D decreased the level of IL-10, increased the TNF-α mRNA, and promoted inflammation-induced cytotoxicity, consistent with the result of inhibiting IL-10 activity with an anti-IL-10 neutralizing antibody.ConclusionsOur study identifies MEF2D as a critical regulator of IL-10 gene expression that negatively controls microglia inflammation response and prevents inflammation-mediated cytotoxicity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12974-015-0258-z) contains supplementary material, which is available to authorized users.

Lack of association between IL-10 -1082G/A polymorphism and chronic periodontal disease in adults.

Because of the complex interaction between periodontal pathogens and the host defense system, periodontitis is considered an inflammatory disorder of bacterial etiology that results in periodontal tissue damage. Genetic mechanisms may interfere with the gene expression of important inflammation mediators, modulating the immunologic response of an individual. In this study, we evaluated the single nucleotide polymorphism -1082G/A in the promoter region of interleukin-10 gene and its relationship with periodontal disease in Central Brazil. We included 36 cases classified according to disease severity (mild, moderate, or severe) and 30 controls. The allelic distribution of the cases was 16 (44%) AG, followed by 13 (36%) GG and 7 (20%) with the genotype AA. In the control group, 13 (43%) presented the genotype AG, 12 (40%) GG and 5 (17%) were classified as AA. The populations examined were in Hardy-Weinberg equilibrium. Analysis of allelic and genotypic frequencies revealed no casual relationship with the presence of genotype G or A and the development of periodontal disease in adults. The single nucleotide polymorphism -1082G/A of the interleukin-10 gene was not predictive of periodontal disease.

IL-18 gene polymorphism in patients with visceral leishmaniasis in East Azarbaijan, Iran.

Visceral leishmaniasis (VL) is a parasitic disease caused by Leishmania species. According to the important role of cellular immunity against VL, this study was directed to determine the frequency of-607A/C and-137G/C genotypes on promoter region of interleukin-18 gene. The study groups included 91 patients with confirmed history of VL, 106 healthy seronegative, and 79 healthy seropositive individuals. All three groups were analyzed by amplification refractory mutation system polymerase chain reaction (ARMS-PCR). The highest rate of -607/A, and -607/C alleles was observed in seronegative individuals (66/67%) and in the patients (72/83%). Allele frequency of -607/C is more than -607/A allele in all groups. In position of-137, frequency of-137/G allele in all groups was more than -137/C. Statistical analysis of distribution of genotypes, did not reveal any significant difference among groups. On the basis of the results, there was no significant association between VL and polymorphism of IL-18 promoter. The results of this study showed that IL-18 gene promoter polymorphisms at positions-607 and-137 are not associated with VL in East Azerbaijan, Iran.

Interleukin‐6 gene‐174 G/C promoter polymorphism and its association with clinical profile of patients with multiple myeloma

In multiple myeloma (MM), the growth and survival of myeloma cells is controlled by interleukin-6 (IL-6), the plasma levels of which is controlled by a guanine/cytosine substitution occurring in position -174 of IL-6 gene promoter region. We studied the occurrence of IL-6-174 G/C polymorphism in patients of MM and correlated the presence of genotypes with serum IL-6 levels and tumor staging. One hundred three patients with MM and 117 age- and sex-matched healthy controls were staged by International Staging System. IL-6 genotypes were evaluated by polymerase chain reaction and restriction enzyme analysis. Serum levels of IL-6 were assessed by enzyme-linked immunosorbent assay. Frequency of GG, GC and CC genotypes did not differ significantly between cases (GG 52%, GC 40%, CC 9%) and controls. The median serum level of IL-6 was significantly higher among the GC genotype versus other genotypes (24 ng/mL, P = 0.007) as compared with the GG versus other genotypes (12 ng/mL, P = 0.001). GC was associated more with stage 3 disease (27%) than was GG (11%) or CC (22% P = 0.001). At position 174 of the IL-6 promoter, patients with GC genotype had higher serum levels of IL-6 and presented with more severe disease compared with patients with GG or CC genotype.