Is There a Link between Thyroid Peroxidase Gene Promoter Polymorphisms and Autoimmune Thyroiditis in the Polish Population?

(1) Autoimmune thyroiditis (AIT) is the most common cause of primary hypothyroidism and one of the most frequent organ-specific autoimmune diseases. Its pathogenesis is polygenic and still requires further research. The aim of the study was to assess, for the first time in the Caucasian population, the role of selected TPO gene promoter polymorphisms (rs2071399 G/A, rs2071400C/T, rs2071402 A/G, and rs2071403 A/G) in the development of AIT. A total of 237 patients diagnosed with AIT and 130 healthy controls were genotyped for four TPO gene polymorphisms, and the results were statistically analyzed to check for the role of these polymorphisms. There were no significant differences in the genotype and allele frequencies of the studied TPO gene promoter polymorphisms between patients and controls (p > 0.05). The haplotype distribution (rs2071400-rs2071402-rs2071403) between the two studied groups was similar for the most common variants (CGA, CAG, TGG). Only a rare haplotype (CGG) occurred more frequently among patients compared to controls (p = 0.04). The studied TPO gene promoter polymorphisms did not show an association with susceptibility to AIT in the Caucasian Polish population, contrary to the results in Japanese patients.

XRCC1 Arg399Gln gene polymorphism and the risk of systemic lupus erythematosus in the Polish population.

It has been shown that DNA repair is reduced in patients with systemic lupus erythematosus (SLE) and that the X-ray repair cross-complementing (XRCC1) Arg399Gln (rs25487) polymorphism may contribute to DNA repair. We evaluated the frequency of the XRCC1 Arg399Gln substitution in patients with SLE (n=265) and controls (n=360) in a sample of the Polish population. The odds ratio (OR) for SLE patients with the Gln/Gln versus Gln/Arg or Arg/Arg genotypes was 1.553 (95% confidence interval [CI]=0.9573-2.520; p=0.0729). OR for the Gln/Gln or Gln/Arg versus Arg/Arg genotype was 1.551 (95% CI=1.122-2.144, p=0.0077). The OR for the 399 Gln allele in patients with SLE was 1.406 (95% CI=1.111-1.779, p=0.0045). There was also a statistically significant p-value of the χ(2) test for the trend observed in the XRCC1 Arg399Gln polymorphism (ptrend=0.0048). We also found a significant contribution of the Gln/Gln or Arg/Gln versus Arg/Arg genotype to the presence of either the malar rash or photosensitivity manifestations of SLE OR=2.241 (1.328-3.781, p=0.0023, pcorr=0.0414). Moreover, the meta-analysis of Taiwanese Han Chinese, Brazilian, and Polish populations showed that the Gln/Gln or Gln/Arg genotype and Gln allele were associated with SLE incidence. OR for the Gln/Gln or Gln/Arg versus Arg/Arg genotype was 1.440 (95% CI=1.15-1.80, p=0.0019) and OR for the Gln allele was 1.27 (95% CI=1.08-1.51, p=0.0051). Our studies may confirm that the XRCC1 Arg399Gln polymorphism may increase the risk of incidence of SLE and the occurrence of some SLE manifestations.

ITGAM Arg77His is associated with disease susceptibility, arthritis, and renal symptoms in systemic lupus erythematosus patients from a sample of the Polish population.

The ITGAM Arg77His (rs1143679) and Ala858Val (rs1143683) polymorphisms have been found to be strong contributors to systemic lupus erythematosus (SLE) development. There are evident population distinctions in terms of SLE distribution and manifestations; therefore, we investigated the distribution of the ITGAM Arg77His and Ala858Val polymorphisms in patients with SLE (n = 154) and control subjects (n = 276) in a sample of the Polish population. We observed that patients with the ITGAM His/His and Arg/His genotypes displayed a 1.811-fold increased risk of SLE incidence (95% confidence intervals [95% CI] = 1.171-2.802, p = 0.0089). Odds ratio (OR) for the homozygous ITGAM His/His genotype was 7.333 (95% CI = 0.8119-66.241, p = 0.0576). We also found that the ITGAM 858Val variant might be a risk factor in the occurrence of SLE; the OR for this allele amounted to 1.458 (95% CI = 1.021-2.080, p = 0.0372). There was an association of the ITGAM His/His and Arg/His genotypes with the occurrence of arthritis OR = 3.486 (95% CI = 1.619-7.508, p = 0.0015). We also observed an association between the ITGAM His/His and Arg/His genotypes and renal symptoms in the course of SLE OR = 2.975 (95% CI = 1.478-5.988; p = 0.0023). Our findings confirmed that there is an association of the ITGAM 77His or 858Val variants with SLE incidence and some clinical manifestation of this autoimmune disorder.

Perforin level in CD4+ T cells from patients with systemic lupus erythematosus.

CD4(+) T cells from patients with systemic lupus erythematosus (SLE) exhibit increased expression of various proteins contributing to defective function of CD4(+) T cells. We evaluated the transcript and protein levels of perforin (PRF1) in CD4(+) T cells from SLE patients (n = 41) and healthy individuals (n = 34). The CD4(+) T cells were obtained by a positive biomagnetic separation system. The amounts of mRNA were determined by reverse transcription and real-time quantitative PCR. The protein levels in the CD4(+) T cells were evaluated by Western blotting analysis. We observed significantly higher levels of PRF1 protein (p = 0.013) in SLE CD4(+) T cells than in controls. There was no significant increase in PRF1 transcript levels (p = 0.908) in CD4(+) T cells from SLE patients as compared to healthy individuals. Moreover, we did not observe a correlation between PRF1 transcript and protein levels in SLE CD4(+) T cells and disease activity expressed by the SLEDAI scale. We confirmed previous observations that demonstrated higher levels of PRF1 protein in CD4(+) T cells from SLE patients. However, we did not find a correlation between PRF1 transcripts and proteins in CD4(+) T cells and SLE disease activity.

Fyn and CD70 expression in CD4+ T cells from patients with systemic lupus erythematosus.

OBJECTIVE: CD4+ T cells from patients with systemic lupus erythematosus (SLE) display defective function that contributes to abnormal activation of B cells and autoantibody production. METHODS: We compared the transcript and protein levels of Fyn and CD70 in CD4+ T cells from patients with SLE (n = 41) and healthy individuals (n = 34). The CD4+ T cells were isolated by positive biomagnetic separation technique. The quantitative analysis of messenger RNA was performed by reverse transcription and real-time quantitative PCR. The protein contents in the CD4+ T cells were determined by Western blotting analysis. RESULTS: We observed significantly higher levels of Fyn (p = 0.03) and CD70 (p = 0.029) transcripts in SLE CD4+ T cells than in controls. There was a significant increase in CD70 protein levels (p < 0.0001), but not Fyn protein levels (p = 0.081) in CD4+ T cells from patients with SLE compared to healthy individuals. In the group with high disease activity [SLE Disease Activity Index (SLEDAI) >/= 9], we observed a significantly higher Fyn protein content than in controls (p = 0.030). There was no correlation between Fyn and CD70 protein levels in SLE CD4+ T cells and disease activity as expressed in the SLEDAI scale. CONCLUSION: We confirmed previous observations of higher expression of CD70 in CD4+ T cells from patients with SLE. Our findings suggest that increased Fyn protein content in CD4+ T cells can be associated with high SLE disease activity.

SDF1-3' G801A polymorphisms in Polish patients with systemic lupus erythematosus.

It has been reported that stromal cell-derived factor-1 (SDF1), currently also designated CXCL12, plays a significant role in the development of nephritis and death in the lupus mice model. Using restriction length fragment polymorphism (RFLP) analysis we assessed the frequencies of SDF1-3' G801A (rs 1801157) polymorphic variants between systemic lupus erythematosus (SLE) patients (n = 150) and controls (n = 300). There were no significant differences in the prevalence of SDF1-3' G801A polymorphic variants in SLE patients and healthy individuals. However, we observed that the SDF1-3' A/A and G/A genotypes (recessive model) contributed to renal manifestations of SLE OR = 3.042 (95% CI = 1.527-6.058, P = 0.002), and the p value stayed statistically significant after Bonferroni correction (p(corr) = 0.032) in SLE patients. We also found an association of the SDF1-3' A/A and G/A genotypes (recessive model) with dermal manifestations of SLE OR = 2.510 (95% CI = 1.247-5.052, P = 0.0122), (p(corr) = 0.1952) but this did not remain statistically significant after Bonferroni correction. Our observations suggest that the SDF1-3' G801A genotype may be associated with some clinical manifestations in patients with SLE.

Asp327Asn polymorphism of sex hormone-binding globulin gene is associated with systemic lupus erythematosus incidence.

Endogenous sex hormones have been observed to have a role in systemic lupus erythematosus (SLE) predisposition. Sex hormone-binding globulin (SHBG) regulates the bioavailability of sex hormones to target tissues. Therefore, we examined the distribution of the SHBG functional polymorphism Asp327Asn (rs6259) in SLE patients (n = 150) and controls (n = 150) in a Polish population. We found a contribution of the SHBG327Asn variant to the development of SLE. Women with the Asp/Asn and Asn/Asn genotypes displayed a 2.630-fold increased risk of SLE (95% CI = 1.561-4.433, P = 0.0003). SHBG has a much higher affinity for testosterone than estradiol, and the SHBG327Asn variant displays a reduction of estradiol clearance. Therefore we suggest that the opposing effects of estrogens and testosterone on the immune system and imbalance in the levels of these hormones in SLE patients can be enhanced by the SHBG327Asn protein variant.

[Influence of genetic factors on development and severity of rheumatoid arthritis--part I]. / Wplyw czynników genetycznych na rozwój i ciezkosc przebiegu reumatoidalnego zapalenia stawów--czesc I.

Both genetic and environmental factors affect susceptibility, severity and probably drugs' efficacy in patients with rheumatoid arthritis (RA). After initial completion of the Human Genome Project on the 16th February 2001, significant progress has been made in identifying other than HLA genome regions linked to the increased RA susceptibility. As an effect several new genes have been recognized as an HLA-independent genetic risk factors of RA. PTPN22 gene polymorphism, C5/TRAF1 genes region polymorphism and TNFAIP3-OLIG3 genes region polymorphism(s) are among newly identified and already confirmed genetic risk factors, whereas STAT 4, CTLA4, PADI4 and IRF5 genes polymorphisms are listed among probable RA development genetic risk factors.

[Influence of genetic factors on development and severity of rheumatoid arthritis--part II]. / Wplyw czynników genetycznych na rozwój i ciezkosc przebiegu reumatoidalnego zapalenia stawów--czesc II.

In our previous paper we have presented recent advances in the knowledge of the genetic risk factors predisposing to rheumatoid arthritis development. In contrast to the progress that has been made in identification of genes responsible for susceptibility to RA, there is still little known about genetic risk factors of the more aggressive and (or) refractory RA. In this part of our review we present a detailed knowledge of the role of genetic factors in severity of RA. Gene polymorphisms of the TNF-alpha gene (-238G/A, -308G/A, +489G/ A), IL-1beta gene (-11C/T, +3953C/T), IL-4 gene (-590C/T), MMP genes (-1607 ins1G/del2G, -1612 ins/del A) and TRAF1/C5 genes region (rs 10818488) are among the most intensively investigated HLA independent genetic risk factors of the severe course of RA.

Sarcoidosis: selected clinical cases.

Two interesting cases of sarcoidosis and associated diagnostic challenges have been presented. Clinical similarities and disparities in the course of sarcoidosis and systemic connective tissue diseases, particularly Sjögren's syndrome, have been addressed. It has been highlighted that all organs, not only the lungs but also for example the liver, can be involved in sarcoidosis. Prompt diagnosis and a proper therapeutic approach are of vital importance.

IL-18 105 A>C polymorphism contributes to renal manifestations in patients with SLE.

Systemic lupus erythematosus (SLE) is a multisystem autoimmune connective tissue disorder characterized by various aberrations including increased production of IL-18. As IL-18 105 A>C polymorphic variants have been linked to increased production of this cytokine, we investigated the prevalence of IL-18 105 A>C (rs549908) polymorphic variants in SLE patients (n = 111) and controls (n = 152). There were no significant differences in the distribution of IL-18 105 A>C polymorphic variants in SLE patients and controls. However, there was a significant association between the IL-18 105 AA genotype (recessive model) and renal manifestations OR = 3.360 (1.523­7.415, P = 0.0039) and the P value remained statistically significant after Bonferroni correction (P corr = 0.0351).Our findings indicate that the IL-18 105 AA genotype variant can contribute to renal manifestations in patients with SLE.

Interleukin-1beta gene (IL-1beta) polymorphisms (SNP -511 and SNP +3953) in thyroid-associated ophthalmopathy (TAO) among the Polish population.

OBJECTIVE: The aim of this study was to evaluate whether the IL-1beta gene could be a genetic marker of the thyroid-associated ophthalmopathy (TAO) development. MATERIALS AND METHODS: The IL-1beta gene polymorphisms at -511 and +3953 regions in 117 TAO patients of Polish origin (ATA/NOSPECS class III or greater) and in 106 controls were studied. RESULTS: We found no significant differences in the frequencies of genotypes and allelic variants for SNP -511 and SNP +3953 between the controls and the studied groups. CONCLUSIONS: No association between the IL-1beta polymorphisms and the TAO existed, so those polymorphisms are not suitable genetic markers for TAO.

Genetic variation in the interleukin-10 gene promoter in Polish patients with systemic lupus erythematosus.

Identification of susceptibility genes in systemic lupus erythematosus (SLE) has recently become a topic of interest. The IL-10 promoter contains three single base-pair substitutions at -627C > A, -854C > T and -1117G > A. These single base-pair substitutions produce three different haplotypes, GCC, ACC and ATA, which affect IL-10 expression. We examined the distribution of -627C > A, -854C > T and -1117G > A IL-10 promoter polymorphisms in patients with SLE (n = 103, women only) and matched controls (n = 300). Despite the higher prevalence of the GCC/GCC, GCC/ATA and ATA/ATA genotypes in SLE patients than in controls, we observed that only GCC/GCC genotype frequency distribution was significant between these groups. We observed that women with the GCC/GCC genotype displayed an approximately twofold increased risk of SLE OR = 2.245 (95% CI = 1.354-3.721, P = 0.0022). We did not find any associations between various genotypes of IL-10 promoter haplotypes and clinical manifestations or autoantibody production in patients with SLE. Our observations indicate that the GCC/GCC promoter genotype may contribute to SLE incidence in Polish patients.

Catalase -262C>T polymorphism in systemic lupus erythematosus in Poland.

It has been reported that reactive oxygen species contribute to pathogenesis of systemic lupus erythematosus (SLE). Catalase (CAT) -330C>T transition, known also as -262C>T, generates three genotypes. The CAT -330CC genotype is associated with a significantly lower CAT expression in comparison to -330CT and -330CT genotypes. Therefore, using restriction length fragment polymorphism analysis, we compared the frequencies of CAT -330C>T polymorphic variants between SLE patients (n = 102) and controls (n = 199). We did not observe significant differences in the prevalence of CAT -330C>T polymorphic variants in SLE patients and controls. However, we found that the CAT -330CC genotype (recessive model) showed a significant association with thrombocytopenia OR = 7.314 (1.977-27.057, P = 0.0017). We also observed that the CAT -330CC genotype (recessive model) is linked with leukopenia OR = 3.232 (1.361-7.676, P = 0.0118), renal manifestations OR = 2.403 (1.085-5.321, P = 0.0471) and presence of anti-snRNP Ab OR = 4.206 (95% CI = 1.405-12.590, P = 0.0131), and anti-Scl-70 Ab, OR = 3.143 (95% CI = 1.171-8.433, P = 0.0343) in SLE patients. Our findings suggest that the CAT -330CC genotype may contribute to some clinical manifestations in patients with SLE.

Manganese superoxide dismutase Ala-9Val mitochondrial targeting sequence polymorphism in systemic lupus erythematosus in Poland.

Systemic lupus erythematosus (SLE) is a chronic and progressive autoimmune disease in which reactive oxygen species contribute to pathogenesis. We analysed the distribution of manganese superoxide dismutase (MnSOD2) 47C>T (Ala-9Val) functional polymorphic variants within the mitochondrial targeting sequence in SLE patients (n = 102) and controls (n = 199). We did not find significant differences in the distribution of MnSOD2 47C>T polymorphic variants in SLE patients and controls. However, we found that MnSOD2 Val/Val genotype (recessive model) showed a significant association with Raynaud's phenomenon, odds ratio (OR) = 12.000 [95% confidence interval (CI) = 2.315-62.193], p = 0.0015. We also found that the MnSOD2 Val/Val genotype contributes to immunologic manifestations, OR = 2.957 (95% CI = 1.207-7.243), p = 0.0222, and anti-dsDNA antibody presence OR = 3.365 (95% CI = 1.364-8.304), p = 0.0107, in patients. Our observations indicate that MnSOD2 Val/Val variant can be linked to some clinical manifestations in patients with SLE.

[Cyclophosphamide in the therapy of rheumatoid arthritis and its complications]. / Rola cyklofosfamidu w leczeniu reumatoidalnego zapalenia stawów i jego powiklan.

Cyclophosphamide has been used in the therapy of rheumatoid arthritis (RA) for nearly fifty years. An experience gathered throughout that time helped to identify indications, profits, restrictions and side effects related to its use. As a result of the progress in RA therapy in the recent years, including introduction of anti - TNF alpha therapy, the importance of cyclophosphamide significantly decreased. However, despite all restrictions related to its use, there are still RA patients for whom cyclophosphamide can be considered as a first choice drug. In this article we review the current knowledge on the issue.

A novel mutation (del 1711 G) in the TBG gene as a cause of complete TBG deficiency.

OBJECTIVE: Inherited thyroxine-binding globulin (TBG) deficiency is caused by mutations in the TBG gene (locus: Xq22.2), which result in defective synthesis or changes in the physical properties or biological function of a protein. DESIGN: We report a novel mutation of the TBG gene causing a complete TBG deficiency in three brothers of Polish origin. DNA was extracted from all of the family members and subjected to sequence analysis. We analyzed the family with a heterozygous mother, a normal father, their three hemizygous affected sons, and their two normal sons. MAIN OUTCOME: Our studies revealed a novel mutation, a single nucleotide deletion (guanine) at position 1711, codon 201 (Asp) in exon 2 (GAC --> AC). This mutation led to a frame shift and premature termination at codon 206, causing a short TBG protein of 205 amino acids (AA) compared to 395 AA of the normal TBG. This new TBG-CD variant was found in the mother and her three affected sons. CONCLUSION: This is a new variant of TBG-CD (TBG-CD-PL Poland) containing 205 AA.

The effect of methylprednisolone pulse treatment on cytokine network in Graves ophthalmopathy.

The etiology of Graves ophthalmopathy (GO), representing the most common extrathyroidal manifestation of Graves disease, is multifactorial. Among multiple genetic, environmental, and endogenous factors, cytokines play a critical role in its etiopathogenesis. We studied an effect of glucocorticoid therapy on the serum IL-6, IL-4, and IL-13 levels in 18 GO patients. All the patients presented euthyroid GO with over 4 points according to the CAS classification (range 4-6; mean 4.94). The patients were treated with methylprednisolone (1 g every second day for three times) followed by 6 months oral prednisone (60 mg/day, with gradual reduction). The clinical examination (Clinical Activity Score and the GO severity by modified NOSPECS classification) and measurement of anti-TPO, anti-TG, anti-TSHR (TRAK), IL-6, IL-4, as well as IL-13 serum levels were performed before, after 2 weeks, and after 6 months of the glucocorticoid therapy. Significant serum IL-6 increases (p < 0.001) and moderate serum IL-4 and IL-13 increases (p < 0.05) were found in GO patients compared with healthy controls. After 2 weeks of the therapy, the serum IL-6 levels decreased in majority of the patients, however after 6-month observation, lower serum IL-6 levels were only in 8 patients who seemed to respond clinically to the therapy (mean value of the Clinical Activity Score decreased from 4.5 before the therapy initiation to 1.25 after 6 months of the glucocorticoid therapy). No changes in IL-4 and IL-13 serum levels during the therapy were observed. Statistical analysis revealed a good correlation between serum IL-6 level and the Clinical Activity Score (p < 0.01). Based on the obtained data, we conclude that IL-6 plays an important role in GO. It seems that IL-6 may serve as a useful factor in the inflammatory events of GO.

Fucosylation of serum alpha1-acid glycoprotein in rheumatoid arthritis patients treated with infliximab.

To analyze fucosylation of alpha(1)-acid glycoprotein (AGP) and to identify relations between AGP fucosylation and clinical and biochemical indices of disease activity in patients with rheumatoid arthritis (RA) treated with monoclonal antitumor necrosis factor (TNF) antibody infliximab, we examined 22 patients with RA who underwent a 54-week treatment with infliximab according to ATTRACT protocol. Blood samples were collected at baseline and before every infusion of infliximab. AGP fucosylation was measured using lectin-binding enzyme-linked immunosorbent assay utilizing fucose-specific lectin Aleuria aurantia (AAL). Moreover, the clinical status/activity, erythrocyte sedimentation rate, serum C-reactive protein (CRP), antitrypsin, alpha(1)-antichymotrypsin, AGP reactivity with concanavalin A, serum C3 and C4 complement components, and serum concentrations of TNF and soluble TNF type 1 and type 2 receptors were determined. In most patients, the fucosylation of AGP decreased rapidly after first infusion of infliximab and remained low during the 54-week therapy (p < 0.001). The decrease in AGP affinity to AAL closely followed changes in clinical and laboratory activity of RA and correlated with pretreatment concentrations of CRP (r = 0.4986, p < 0.05) and TNF (r = 0.5181, p < 0.05). The fucosylation of AGP can be a part of a negative feedback loop regulating migration of inflammatory cells and collagenase-3 activity in RA. The decrease in AGP fucosylation accompanied by improvement in clinical and biochemical parameters of RA could possibly reflect reduced migration of inflammatory cells to inflamed joints and AGP-mediated inhibition of collagenase-3 as a response to infliximab treatment.

[Amyloidosis--life threatening complication in rheumatoid arthritis patients]. / Skrobiawica--zagrazajace zyciu powiklanie u chorych na reumatoidalne zapalenie stawów.

Rheumatoid arthritis (RA) is a chronic, inflammatory, autoimmune disease leading to severe disability. A secondary amyloidosis (AA amyloidosis) affecting RA patient is a life threatening clinical complication of the illness. The most common symptoms of secondary amyloidosis include: proteinuria, erythrocyturia, abdominal pain and chronic diarrhoea. It is essential to carry out regular screening tests, especially abdominal fat tissue biopsy, to early diagnose and properly manage patients with the condition. Effective anti-inflammatory therapy of RA and eradication of coexisting infections seem to be the best way to decrease the risk of development and prevent progression of the secondary amyloidosis.