CD3Z polymorphisms and promoter hypermethylation in dermatomyositis - the role of cytosine-phosphate-guanine-related single nucleotide polymorphisms.

BACKGROUND: Decreased expression of the T cell receptor (TCR) ζ-chain has been reported in autoimmune diseases. Recent evidence suggests that this deficiency may be due to polymorphisms in the CD3Z (CD247) gene and/or due to promoter hypermethylation. METHODS: Altogether 131 subjects - 36 with dermatomyositis (DM) and 95 healthy controls were genotyped for rs1052230 G > C and rs1052231 T > A polymorphisms using TaqMan assay. The rs840015 G > A polymorphism was analyzed by direct sequencing. The promoter methylation status was analyzed by Sanger sequencing of bisulfite converted DNA. RESULTS: The rs1052230GC genotype and C allele and the rs1052231TA genotype and T allele were found to correlate with photosensitivity as well as the rs1052230C/rs1052231T haplotype. The rs1052231TA genotype was found to be associated with cutaneous disease. The rs840015GG genotype was found increased among patients with DM, leading to increased OR 2.4. On the contrary, the rs840015GA genotype appeared to be protective for the development of DM. From the 11 cytosine-phosphate-guanine (CpG) islands analyzed, only the 8th island showed a difference in its methylation due to the polymorphism rs840015 G > A within this island, as our results suggest. In this way the presence of AA genotype led to no methylation and the presence of the GG genotype was associated with hemimethylation. CONCLUSION: The CD247 rs1052230 G > C and rs1052231 T > A polymorphisms appeared to have a disease-modifying role. The rs840015GA genotype being associated with reduced methylation has a protective role for the development of dermatomyositis and our results suggest that CpG related single nucleotide polymorphisms may play an important role in autoimmunity.

The role of IL-17 rs2275913, IL-17RC rs708567 and TGFB1 rs1800469 SNPs and IL-17A serum levels in patients with lupus nephritis.

Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease and polymorphisms in the cytokine genes and their receptors are thought to influence its development. The aim of this case-control study was to investigate the association of the IL-17A rs2275913, IL-17RC rs708567 and TGFB1 rs1800469 polymorphisms with SLE, its clinical manifestations and the polymorphisms influence on the IL-17A serum levels. Altogether 59 SLE patients with lupus nephritis and 95 healthy controls were genotyped by TaqMan assay. Serum levels were determined by Human IL-17A Platinum ELISA kit. From the studied polymorphisms, only TGFB1 T allele was found to be associated with SLE. Within the patient group, IL-17A GG genotype and TGFB1 -509T allele showed an association with the neurological disease and IL-17RC CC genotype appeared to be associated with lupus arthritis. The IL17A serum levels in the SLE and control groups (7.24 pg/ml and 5.76 pg/ml, respectively) did not show any statistical difference. A weak correlation between IL17A levels and SLEDAI-2K was observed. Our results indicate that IL-17A rs2275913, IL-17RCrs708567 and TGFB1 rs1800469 polymorphisms might play a role in the susceptibility and the clinical manifestations of SLE and IL-17A serum levels should be monitored in the course of the disease. The identification of subsets of SLE with an IL-17-driven disease could improve the therapeutic approach leading to more precise personalized treatment.

XRCC1 variants do not represent a risk for dermatomyositis and systemic lupus erythematosus in Bulgarian patients.

INTRODUCTION: Systemic lupus erythematosus (SLE) and dermatomyositis (DM) share a similar pathogenesis, and genetic, hormonal, and environmental factors are known to trigger the autoimmune process. The X-ray repair cross-complementing genes (XRCC1 and XRCC3) are known to play a central role in mammalian DNA repair processes. Evidence suggests that impaired DNA repair efficiency is implicated in the development of autoimmune diseases. This case-control study investigates the association between the XRCC1 Arg194Trp (C>T) and Arg399Gln (G>A) polymorphisms and the susceptibility to DM and SLE in Bulgarian patients. METHODS: Altogether 88 patients, 55 with SLE and 33 with DM, and 94 unrelated healthy controls were included in this study. RESULTS: None of the polymorphisms showed an association with SLE, DM, or their clinical parameters. The allele and genotype frequency of the two single nucleotide polymorphisms was similar to those found in other healthy Caucasian populations. CONCLUSIONS: Our results indicate that the XRCC1 rs1799782 Arg194Trp and rs25487 Arg399Gln polymorphisms do not play a role in the susceptibility to SLE and DM.

Genetically Determined TNF-α Secretion Influences the Development of Dermatomyositis and Systemic Lupus Erythematosus.

BACKGROUND: Abnormal secretion of TNF-α is known to play a role in the pathogenesis of dermatomyositis and systemic lupus erythematosus. MATERIALS AND METHODS: In the present study we have analyzed the concentrations of TNF-α in the sera of 30 patients with systemic lupus erythematosus (SLE), 28 with dermatomyositis (DM) and 30 healthy controls by standard ELISA tests. RESULTS: We have found that -308A allele increases TNF-α secretion, while -1031C and -863A alleles decrease it. The -857C/T and 489G/A polymorphisms appeared in strong linkage disequilibrium (D'=0.93) but they did not seem to affect TNF-α secretion. CONCLUSION: TNF-α polymorphisms play a significant role in its secretion and influence the development of DM and SLE.

The Role of CD247 Polymorphisms in Bulgarian Patients with Systemic Lupus Erythematosus.

Decreased expression of the TCR ζ-chain has been reported in several autoimmune and inflammatory diseases. Recent evidence suggests that this deficiency may be due to polymorphisms in the CD247 gene. A total 52 patients with systemic lupus erythematosus (SLE) and 95 healthy controls of Bulgarian ethnicity were genotyped for 837C>G, rs1052230, 844A>T, and rs1052231 using a TaqMan genotyping assay. None of the two polymorphisms appeared associated with the diseases. On the other hand, we have found that the -837GG genotype and the G allele were associated with hematological disease. The -844AA genotype and the A allele appeared associated with the hematological disease as well. The -843AA genotype and the A allele were found to be associated with antinuclear antibody (ANA) tests and immunological disease. An association was found between the -837G allele and arthritis. The AG haplotype was found to be associated with hematological disease, ANA, and immunological disease. Our preliminary data confirm the previous findings that the CD247 polymorphisms are mainly associated with the clinical outcome of the disease and less with susceptibility.

Monoclonal Antibody Drugs for Systemic Lupus Erythematosus.

Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disease which engages most of the immune cells in its development. Various studies concerning the application of antibodies against TNF-α, BlyS, CD20, CD22, IL-6R and complement factors in treatment of SLE have been recently conducted and in spite of the good results reported by some of them, no definite conclusion on their risk-benefit profile can be drawn. The current review summarizes the results obtained in the field and reveals the perspectives for the development of new and more effective strategies for SLE treatment in combination with other immunomodulating drugs.

IL-1RN VNTR Polymorphism in Adult Dermatomyositis and Systemic Lupus Erythematosus.

Polymorphisms in the cytokine genes and their natural antagonists are thought to influence the predisposition to dermatomyositis (DM) and systemic lupus erythematosus (SLE). A variable number tandem repeat (VNTR) polymorphism of 86 bp in intron 2 of the interleukin-1 receptor antagonist (IL-1RN) gene leads to the existence of five different alleles which cause differences in the production of both IL-1RA (interleukin-1 receptor antagonist) and IL-1ß. The aim of this case-control study was to investigate the association between the IL-1RN VNTR polymorphism and the susceptibility to DM and SLE in Bulgarian patients. Altogether 91 patients, 55 with SLE and 36 with DM, as well as 112 unrelated healthy controls, were included in this study. Only three alleles were identified in both patients and controls ((1) four repeats, (2) two repeats, and (3) five repeats). The IL-1RN*2 allele (P = 0.02, OR 2.5, and 95% CI 1.2-5.4) and the 1/2+2/2 genotypes were found prevalent among the SLE patients (P = 0.05, OR 2.6, and 95% CI 1-6.3). No association was found between this polymorphism and the ACR criteria for SLE as well as with the susceptibility to DM. Our results indicate that the IL-1RN VNTR polymorphism might play a role in the susceptibility of SLE but not DM.

Role of the promoter polymorphism IL-6 -174G/C in dermatomyositis and systemic lupus erythematosus.

The promoter polymorphism -174G/C within the interleukin-6 gene (IL-6) has been reported to have a functional importance through the modulation of IL-6 gene expression in vitro and in vivo. IL-6 is thought to play an important role in autoimmune diseases and the effect of its receptor inhibitor-tocilizumab-has been recently studied. The aim of this case-control study was to investigate the association between the interleukin-6 -174G/C single nucleotide polymorphism and the susceptibility to dermatomyositis (DM) and systemic lupus erythematosus (SLE) in Bulgarian patients. Altogether, 87 patients-52 with SLE and 35 with DM-as well as 80 unrelated healthy controls were included in this study. All of them were analyzed by restriction fragment length polymorphism analysis (RFLP). The GG genotype and the G allele appeared to be associated with SLE, especially in women. None of the genotypes showed an association with DM. However, the G allele appeared to be associated with muscle weakness and it is a risk factor for elevated muscle enzymes. Our results indicate that IL-6 -174G/C polymorphism might be associated with the susceptibility to SLE especially in women. Although it is not associated with DM, it seems that IL-6 -174G/C polymorphism could modulate some clinical features in the autoimmune myopathies.

Association of tumor necrosis factor α (TNF-α) and interleukin 10 (IL-10) gene polymorphisms in dermatomyositis patients: a pilot study.

TNF-α and IL-10 single nucleotide polymorphisms have been implicated in various autoimmune diseases but the results are still quite controversial. This case-control study aimed to investigate the association between six TNF-α and five IL-10 polymorphisms with dermatomyositis. The -857CC and +489GG genotypes showed a weak association with dermatomyositis when the analysis was carried out for the whole cohort but they appeared to be a significant risk factor for the development of dermatomyositis in women. The TNF-α -1031CC genotype was found only among dermatomyositis patients. The TNF-α -1031C/-863C/-857C-308G/+489G haplotype showed a significant association with dermatomyositis in women. The IL-10 -3575TT genotype and T allele showed an association with dermatomyositis. The frequency of the IL-10 -2763CC genotype and C allele was higher among dermatomyositis patents and it was associated with an increased OR. Haplotype analysis showed an association between the IL-10 -3575T/-2763C haplotype and dermatomyositis. In conclusion, our results indicate that both TNF-α and IL-10 polymorphisms are associated with the development of dermatomyositis in Bulgarian patients.

Association of TNF-α polymorphisms with adult dermatomyositis and systemic lupus erythematosus in Bulgarian patients.

BACKGROUND: Single-nucleotide polymorphisms (SNPs) of tumor necrosis factor-alpha (TNF-α) have been implicated in various autoimmune diseases; however, the results are quite controversial, and there is still no widely accepted opinion about their role in the pathology of the autoimmune diseases. This is a pilot study to investigate the association of six SNPs of the TNF-α gene with the risk of adult dermatomyositis (DM) and systemic lupus erythematosus (SLE) in Bulgarian patients. MATERIALS AND METHODS: Twenty-seven patients with DM and 27 with SLE were included in this study. Genomic DNA was extracted from the peripheral blood, and six SNPs (-1031T/C, -863C/A, -857C/T, -308G/A, -238G/A, +489G/A) were selected for investigation by polymerase chain reaction-restriction fragment length polymorphism analysis. RESULTS: We found association between the TNF-α-1031CC genotype and SLE (P = 0.025) and tendency for association with DM (P = 0.0876). The association appeared even stronger in the female patients with SLE (P = 0.024) and DM (P = 0.067). The TNF-α-857GG genotype shows weak association with SLE (P = 0.097, OR 2.06, 95% CI 0.81-5.29) when analyzed for the whole group, but it appeared significantly associated with SLE in women (P = 0.048, OR 3.23, 95% CI 0.93-11.14). The -863C allele showed association with arthritis in patients with SLE (P = 0.008). The haplotype analysis revealed a significant association between TNF -1031C/-863C/-857C/-308G/+489G haplotype with both DM (P = 0.022) and SLE (P = 0.007) in women. CONCLUSIONS: The TNF-α polymorphisms are associated with increased relative risk mainly for SLE, particularly in women, while their role for DM is less evident and needs further analysis in an enlarged sample cohort.

A vanadium-dependent bromoperoxidase in the marine red alga Kappaphycus alvarezii (Doty) Doty displays clear substrate specificity.

Bromoperoxidase activity was initially detected in marine macroalgae belonging to the Solieriaceae family (Gigartinales, Rhodophyta), including Solieria robusta (Greville) Kylin, Eucheuma serra J. Agardh and Kappaphycus alvarezii (Doty) Doty, which are important industrial sources of the polysaccharide carrageenan. Notably, the purification of bromoperoxidase was difficult because due to the coexistence of viscoid polysaccharides. The activity of the partially purified enzyme was dependent on the vanadate ion, and displayed a distinct substrate spectrum from that of previously reported vanadium-dependent bromoperoxidases of marine macroalgae. The enzyme was specific for Br- and I- ions and inactive toward F- and Cl-. The K(m) values for Br- and H2O2 were 2.5x10(-3) M and 8.5x10(-5) M, respectively. The halogenated product, dibromoacetaldehyde, that accumulated in K. alvarezii was additionally determined.

Sterol and lipid composition of three Adriatic Sea sponges.

The sterol and fatty acid composition of three Adriatic Sea sponges (Geodia cydonium and two unidentified Tedania sp.), collected at the same time and same place, was established. Twenty-four sterols and forty fatty acids were identified. The identical ecological conditions, including the diet, allowed us to apply the results obtained for taxonomical conclusions, based on the biodiversity of the investigated sponges. On the basis of the sterol composition they can be separated into two groups: Tedania and Geodia sponges. The sterol and fatty acid composition indicates that the two investigated Tedania samples might be different species or subspecies.

Comparative investigation on sterols from some Black Sea red algae.

The sterol composition of five Black Sea red algae: Callithamnion granulatum, Ceramium rubrum, Ceramium elegans, Laurencia papillosa and Laurencia coronopus was obtained. Monohydroxy sterols and their oxidized derivatives have been identified. The results obtained were compared with some recent data for the composition of sterols in other Black Sea red algae, which occupy different evolutionary positions. On the basis of their composition, some conclusions about the chemotaxonomy and chemoevolution of the species investigated have been proposed.

Synthesis of some phenylpropanoid monoglycerides via the Mitsunobu protocol.

Natural monoglycerides of cinnamic, ferulic and p-coumaric acids were synthesized in good to high overall yields from isopropylidene glycerol via the Mitsunobu reaction and further deprotection of the corresponding acetonides with Amberlyst 15. The method avoids the need of protection of the phenolic hydroxyls. During the Mitsunobu esterifications a strong influence of the acid strength on the efficacy and outcome of the reaction was observed.

Chemical composition of Corallina mediterranea Areschoug and Corallina granifera Ell. et Soland.

The composition of sterols, volatiles and some polar compounds from three Corallina samples (C. granifera and C. mediterranea from the Black Sea and C. mediterranea from the Mediterranean Sea) was established. The sterol composition of the Black Sea samples was similar but it differs from that of the Mediterranean sample. The composition of the volatiles was very complex. The main groups of constituent were hydrocarbons, alcohols, carbonyl compounds, acids and their esters, terpenes. The composition of the polar components, soluble in n-butanol, was also established. There were some differences in the chemical composition of the two Black Sea species, which may be due to the biodiversity between them, while the differences in the composition of the two C. mediterranea samples could be due to the differences in the environment (salinity, temperature, pollution, etc.).

Lipophylic compounds from Euphorbia peplis L.--a halophytic plant from the Bulgarian Black Sea coast.

The chemical composition of the lipophylic fraction from the halophytic plant Euphorbia peplis L. was investigated. Compared to other terrestrial higher plants an increase of triacylglycerols and especially of glycolipids was observed. The main phospholipid was phosphatidyl choline, followed by almost equal concentrations of phosphatidyl ethanolamine and phosphatidyl glycerol. A relatively high concentration of phosphatidic acids (6.5% of the total phospholipids) was found. The main sterol appeared to be sitosterol and significant amounts of tetracyclic triterpene alcohols were found. The composition of the volatile compounds is relatively simple and only one chlorinated compound, identified as 2,2-diethoxy-1-chloroethane, was found. There was a strong toxicity of the total lipophylic extract towards Artemia salina.

Chemical composition of Cystoseira crinita Bory from the Eastern Mediterranean.

The chemical composition of the brown alga Cystoseira crinita Bory from the Eastern Mediterranean was investigated. Fourteen sterols have been identified, five of them for the first time in algae. The structure of one new sterol was established. The origin of seven sterols with short side chains was discussed. In the volatile fraction 19 compounds and in the polar fraction 15 compounds were identified. The main lipid classes were isolated and their fatty acid composition was established.