Clinical relevance of thyroid-stimulating autoantibodies in pediatric graves' disease-a multicenter study.

CONTEXT AND OBJECTIVE: The incidence of TSH receptor (TSHR) stimulating autoantibodies (TSAbs) in pediatric Graves' disease (GD) is controversial. This large, multicenter study evaluated the clinical relevance of TSAbs in children with GD both with Graves' orbitopathy (GO) and without orbital disease. DESIGN: We conducted a cross-sectional retrospective study. SETTING: Sera were collected in seven American and European academic referral centers and evaluated in a central laboratory. PATIENTS AND SAMPLES: A total of 422 serum samples from 157 children with GD, 101 control individuals with other thyroid and nonthyroid autoimmune diseases, and 50 healthy children were studied. MAIN OUTCOME MEASURES: TSAbs were measured using a novel, chimeric TSHR bioassay and a cAMP response element-dependent luciferase. TSH binding-inhibitory Ig (TBII) and parameters of thyroid function were also determined. RESULTS: In 82 untreated children with GD, sensitivity, specificity, and positive and negative predictive values for TSAb and TBII were: 100 and 92.68% (P = .031), 100 and 100%, 100 and 100%, and 100 and 96.15%, respectively. TSAb and TBII were present in 147 (94%) and 138 (87.9%) of the 157 children with GD (P < .039), respectively; and in 247 (94%) and 233 (89%) of the 263 samples from this group (P < .0075), respectively. In children with GD and GO, TSAb and TBII were noted in 100 and 96% (P < .001), respectively. Hyperthyroid children with GD and GO showed markedly higher TSAb levels compared to those with thyroidal GD only (P < .0001). No significant differences were noted for TBII between the two groups. After a 3-year (median) medical treatment, the decrease of TSAb levels was 69% in GD vs 20% in GD and GO (P < .001). All 31 samples of euthyroid children with GO were TSAb positive; in contrast, only 24 were TBII positive (P = .016). All children with Hashimoto's thyroiditis, nonautoimmune hyperthyroidism, type 1 diabetes, and juvenile arthritis and the healthy controls were TSAb and TBII negative. CONCLUSIONS: Serum TSAb level is a sensitive, specific, and reproducible biomarker for pediatric GD and correlates well with disease severity and extrathyroidal manifestations.

Monthly use of a real-time continuous glucose monitoring system as an educational and motivational tool for poorly controlled type 1 diabetes adolescents.

PURPOSE: Experience with the use of real-time continuous glucose monitoring systems (RT-CGMS) in teenagers with type 1 diabetes mellitus (T1DM) is limited. We aimed to assess the possibility of glycaemic control improvement and to characterize the group of adolescents, who may gain long-term benefits from the use of the RT-CGMS. METHODS: Forty T1DM patients, aged 14.6 ± 2.1 years, with diabetes duration 7.4 ± 3.6 years and initial HbA1c 9.3 ± 1.5% were recruited. The analysis was based on one-month glucose sensors use, combined with the thorough family support. Patients were analysed in groups according to baseline HbA1c: below and above 7.5%, and 10.0%. Comparison between patients with or without improvement in HbA1c after 3-month follow-up was also performed. Patients' satisfaction based on the questionnaire was assessed. RESULTS: HbA1c level in entire study group decreased after three months, from 9.3 ± 1.0% to 8.8 ± 1.6% (P<0.001). In the group with HbA1c improvement, reduction was the highest: 9.0 ± 1.3% vs. 8.0 ± 1.2% (P<0.001). Only the group with initial HbA1c>10% did not achieve significant improvement: 11.2 ± 0.5% vs. 10.9 ± 1.1 (P=0.06). In satisfaction questionnaire the lowest scores (negative opinion) were reported by group of patients with initial HbA1c above 10%, while the highest scores (positive opinion) were found in the group with improvement of HbA1c after 3 month follow-up. CONCLUSION: Short-term use of CGMS RT, united with satisfaction questionnaire, performed in poorly controlled teenagers with T1DM, can be useful in defining the group of young patients, who can benefit from long-term CGMS RT use in metabolic control improvement.

Syncope in children and adolescents living in north-eastern Poland - scope of causes.

PURPOSE: To investigate the diagnostic yield of different tests and asses the scope of causes in children referring to the hospital with a syncope in north-eastern Poland. METHODS: A review of 386 consecutive patients (age 2-18 years) living in north-eastern Poland presenting to the cardiology department with a new onset syncope (which appeared to be neurally mediated by history) was undertaken. The patients underwent physical examination, laboratory tests, electrocardiography, 24-holter monitoring, head-up tilt-test, exercise test, echocardiography and electroencephalography. All the tests were performed in most of the patients, without ending the diagnostics after finding the first probable cause of loss of consciousness. RESULTS: 229 potential causes of syncope were found in 191 patients (49.4%), with 2 possible causes in 32 patients and 3 potential causes in 3 patients. The top 3 tests with the highest diagnostic yield were: head-up tilt test (41.4%), 24-holter monitoring (14.5%) and echocardiography (8.4%). Electroencephalography was useful in 3 patients (1.5%) and exercise test did not help in any patient. CONCLUSION: In some children more than one potential cause of syncope was diagnosed. That might confirm multiple factors' causality of syncope. Head-up tilt-test has the highest diagnostic yield in children with syncope. If the diagnostic protocol is not very strict, one might find some crucial conditions in about 8.3% of patients.

Cardiovascular risk in nonobese hypertensive adolescents: a study based on plasma biomarkers and ultrasonographic assessment of early atherosclerosis.

The objective of this study was to investigate the vascular status, left-ventricular mass and biomarkers of endothelial activation in hypertensive (HT) adolescents, with particular attention to comparing nonobese with obese patients. Seventy-nine newly diagnosed HT adolescents aged 15.1±2.1 years (divided into 34 nonobese and 45 obese) were compared with 35 healthy volunteers. Intima-media thickness (IMT), flow-mediated dilation (FMD) and left-ventricular mass index (LVMi) were determined using ultrasound. Adhesion molecules and inflammatory interleukins (ILs), together with lipids and insulin resistance (HOMA), were also studied. HT obese adolescents had higher triglycerides, HOMA, and elevated levels of interleukin-6, tumor necrosis factor-α, soluble intercellular adhesion molecule-1 and soluble E-selectin compared with controls and nonobese HT patients. FMD was lower in HT groups (8.5±4.5% in nonobese, P=0.004; 8.1±4.9%, P=0.01 in obese vs 12.5±4.9%; in control), and IMT was higher (0.52±0.06 mm, P<0.001 in nonobese; 0.54±0.05 mm, P<0.001 in obese vs 0.42±0.05 mm in control). Higher LVMi was found in both HT groups, with the highest value in the nonobese group being 37.8±5.3 g m(-2.7) vs 28.4±5.3 g m(-2.7) in controls (P=0.003). In conclusion, nonobese HT adolescents had the same early cardiovascular deteriorations assessed ultrasonographically as their obese HT peers, although metabolic alterations and endothelial activation measured as plasma biomarkers were more pronounced in obese individuals. The potential mechanisms of early atherosclerosis in nonobese HT adolescents need further evaluation in prospective studies because these factors may differ considerably from those found in young obese individuals with HT.

Lower proportions of CD4+CD25(high) and CD4+FoxP3, but not CD4+CD25+CD127(low) FoxP3+ T cell levels in children with autoimmune thyroid diseases.

The essence of autoimmune thyroid disease (AITD) is loss of tolerance of own tissues caused by malfunction of T lymphocytes, which affects the production of antibodies reacting with particular cell structures and tissues. Foxp3(+) regulatory T cells (Tregs) take part in the regulation of immune response and play a leading role in developing immune tolerance through active suppression. The aim of the study was to estimate the expression of CD4+CD25(high), CD4+CD25+CD127(low)FoxP3(+) and CD4+ FoxP3 T cells in patients with Graves' disease (GD) (n = 24, median age 15.5 years), in patients with Hashimoto's thyroiditis (HT) (n = 30, median age 15 years) in comparison with sex- and age-matched healthy control subjects (n = 30, median age 15 years). Polychromatic flow cytometry using a FACSCalibur (BD Biosciences) cytometer was applied to delineate T regulatory cell populations. In untreated patients with Graves' disease and HT we observed a significant decrease in CD4+FoxP3 (p < 0.001, p < 0.01) and CD4+CD25(high) (p < 0.016, p < 0.048) T lymphocytes as compared to the healthy control subjects. After 6-12 months of L-thyroxine therapy in HT cases these phenotypes of Tregs were normalized, yet no such changes were observed during GD therapy. The analysis of CD4+CD25+CD127(low)FoxP3+ T cells in the peripheral blood revealed comparable percentages of these cells in patients with thyroid autoimmune diseases to the controls. We conclude that the reduction number of Tregs with CD4+CD25(high) and CD4+FoxP3 phenotype suggests their role in initiation and development of autoimmune process in thyroid disorders.

The association of the FTO rs9939609 polymorphism with obesity and metabolic risk factors for cardiovascular diseases in Polish children.

The phenomenon of excessive body mass increase in the general population of children and adults in the past few decades and has been attributed to environmental changes, especially when superimposed on a predisposing genetic background. The fat mass and obesity-associated (FTO) gene has recently become one of the most extensively investigated genes associated with body mass. The aim of the study was to investigate the association of the FTO rs9939609 T>A single-nucleotide polymorphism with selected anthropometric and metabolic parameters and blood pressure in a large population of Polish children. A total of 968 children aged 4 to 18 years were included in the study. Genotyping was performed using a TaqMan assay. The rs9939609 marker was associated with standardised (standard deviation scores, SDSs) values of body mass, height, body mass index (BMI), waist circumference, hip circumference and arm circumference. When we compared normal-weight with obese children we observed a strong recessive predisposing effect of the A-allele (OR=2.11 95% CI: 1.50-2.99, p=2.23 x 10(-6) for AA vs. TT+AT). Univariate analysis revealed associations of the FTO gene variants with the values of blood pressure, triglycerides, fasting glucose and HOMA insulin resistance index. After taking into account SDS BMI only the association with HOMA remained statistically significant. The FTO gene polymorphism may partially explain the predisposition to obesity in the population of Polish children. The potential effect on the remaining cardiovascular risk factors seems indirect and dependent on BMI changes. The polymorphism may be independently associated with insulin resistance.

Generation of T regulatory cells in children with newly diagnosed type 1 diabetes mellitus.

UNLABELLED: There is increasing evidence that T-regulatory (Treg) cells could be used to prevent or cure autoimmune diseases including type 1 diabetes mellitus (T1DM). The aim of the present study was to verify the hypothesis that functional Treg cells can be generated from conventional T-cells separated from a small amount of peripheral blood of children with newly diagnosed T1DM (N=25). METHODS: CD4(+)CD25(-) cells were cultured with Treg expander (CD3/CD28) and IL-2 for generating de novo Treg cells. The assessment of the expression of selected genes and proteins critical to Treg function and the proliferation assays were performed with the use of real-time RT-PCR and flow cytometry. RESULTS: After a 4-week stimulation with Treg expander and IL-2, the percentage of T-regulatory cells was significantly higher compared to the cells treated with medium alone (with no difference between diabetic and control children). However, we found some disturbances in the gene expression at mRNA level for molecules crucial for T-reg function. The induced Tregs from diabetic and control children were fully functional as assessed in proliferation assays. IN SUMMARY: Despite some disturbances at mRNA level in the critical gene expression, the suppressive properties of induced Treg cells from diabetic and control children were effective.

Identification of chosen apoptotic (TIAR and TIA-1) markers expression in thyroid tissues from adolescents with immune and non-immune thyroid diseases.

The aim of this study was to estimate sodium iodide symporter (NIS) and thyroid peroxidase (TPO) expression in thyrocytes from patients with GD and no-toxic multinodular goitre (NTMG) in relationship with apoptotic (TIAR and TIA-1) markers. The investigation was performed on thyroid cells isolated from postoperation thyroid tissues from 15 patients aged 12-21 years old with GD and 15 cases aged 13-21 years old with NTMG. Detection of NIS and TPO was performed by immunohistochemistry. Analysis of apoptotic markers in thyroid tissues was performed using antibodies to TIAR and TIA-1 by Western Blot and immunohistochemistry. Identification of proapoptotic TIAR and TIA-1 molecules in the thyroid tissues revealed a higher expression of both proteins in patients with Graves' disease (+++; +, respectively) in comparison to patients with NTNG (+; 0). In addition, TIAR expression was detected in three bands [p50, p42, p38 (kDa)] and TIA-1 in two bands [p22, p17 (kDa)]. using Western Blot test in patients with thyroid autoimmune diseases. In patients with NTNG expression of both apoptotic proteins was lower and identified in single bands: 42 (kDa) for TIAR and 17 (kDa) for TIA-1. The analysis of expression of NIS and TPO in thyroid follicular cells was higher in patients with Graves' disease in compared to their detection in patients with NTMG. In addition, degree of thyroid antigen expression positive correlated with amount of proapoptotic markers (TIAR, p<0.001; TIA-1, p<0.025 for NIS; TIAR, p<0.012 for TPO). We conclude that elevated expression of NIS and TPO in Graves' disease is associated with higher stimulation and activation of apoptosis in thyroid follicular cells during autoimmune process.

Relationship between metabolic parameters and thyroid hormones and the level of gastric peptides in children with autoimmune thyroid diseases.

Overweight and diseases connected with it are increasing problems in children and adults. We often observe change of weight in thyroid disease. It is emphasized that changes in hormones such as peptide levels are in close relationship with regulation of body mass: ghrelin increases appetite and in effect increases body mass, but obestatin decreases appetite and weight. The aim of the study was to analyze the relationship between lipid-carbohydrate metabolism parameters and thyroid hormones and the level of gastric peptides (ghrelin and obestatin) in young patients with Graves' disease, Hashimoto's thyroiditis and in children with simple goiter. The study group formed 78 patients suffering from Graves' disease (29 girls and 2 boys; aged from 6 to 21 - mean 15,2 yrs) and Hashimoto's thyroiditis (29 girls and 3 boys; aged from 9 to 18--mean 14.5 yrs). The control group consisted of children with simple goiter--13 girls and 2 boys; aged from 9 to 18 --mean 14.8 yrs. In all patients, ghrelin and obestatin levels were analyzed by the RIA method (Phoenix Pharmaceuticals, USA). In children and adolescents with untreated Graves' disease we found higher levels of insulin and HOMA-IR index compared to the group of children with simple goiter (34 +/- 8 microIU/mL vs 15 +/- 5; p < 0.03; 7.3 +/- 1.2 vs 3 +/- 0.3, p < 0.03). No significant correlations were observed of gastric hormones with antithyroid antibodies, lipids or h-CRP in patients with untreated hyperthyroidism and subclinical hypothyroidism. Positive correlation was noted of insulin and glucose levels and HOMA-IR index with ghrelin level in children with newly diagnosed Graves' disease (r = 0.109, p < 0.045; r = 0.176, p < 0.036; r = 0.174, p < 0.037). The correlation was also positive between obestatin level and HOMA-IR index in children with subclinical hypothyroidism in the course of Hashimoto's thyroiditis (r = 0.497, p < 0.011). We also examined the relationship between BMI, thyroid hormones and the level of gastric peptides. In untreated GD patients, ghrelin level exhibited a significant negative correlation with fT3 and fT4 (r = -0.38, p < 0.041; r = -0.459, p < 0.012) and positive with TSH (r = 0.38, p < 0.041) and BMI (r = 0.8, p < 0.01). In conclusion, we suggest that the disturbances in carbohydrate parameters in thyroid diseases have an essential effect on change of hormone-controlled appetite: ghrelin (in hyperthyroidism) and obestatin (in Subclinical hypothyroidism).

Expression of Bcl-2 family proteins in thyrocytes from young patients with immune and nonimmune thyroid diseases.

The Bcl-2 family proteins that control homeostasis of cells play an important role in apoptosis. This group consists of antiapoptotic (Bcl-2, Bcl-XL) and proapoptotic (Bcl-2 associated protein X, Bax; B-cell homologous antagonist/killer, Bak) molecules. In the thyroid, abnormal apoptotic activity may be involved in a variety of diseases such as autoimmune thyroid diseases. The aim of the current study was to estimate the expression of pro- and antiapoptotic proteins in thyroid tissues from young patients with Graves' disease (GD), nontoxic nodular goiter and toxic nodular goiter using Western Blot and immunohistochemistry. Identification of the antiapoptotic Bcl-2 and Bcl-XL molecules in the thyrocytes revealed higher expression of both proteins in patients with GD (assessed as +++/++ and ++/+, respectively). In adolescents with toxic and nontoxic nodular goiter, this expression was lower (Bcl-2 ++/+ , ++/+; Bcl-XL +, +). The tissue material was additionally subjected to Western Blot analysis, which in GD patients showed the presence of Bcl-2 and Bcl-XL in one band p26 kDa. In patients with toxic and nontoxic nodular goiter, the intensity of expression for these two antiapoptotic proteins was lower (referred to band 26 kDa for Bcl-2 and Bcl-XL). Identification of the proapoptotic proteins Bax and Bak revealed their predominance in thyrocytes of GD patients (+, ++/+, respectively) as compared to patients with toxic and nontoxic nodular goiter (0/+, 0/+ for Bax and 0/+, 0/+ for Bak). In GD patients, Western Blot analysis showed Bax expression in one band 21 kDa and Bak in two bands p50, p24 kDa. In patients with nodular goiter, the degree of expression of both proapoptotic proteins was lower and referred to band 21 kDa for Bax (toxic and nontoxic goiter) and 24 kDa for Bak (toxic goiter only). Patients with GD showed a statistically significant correlation between Bcl-2 expression and antibodies against receptor for thyroid stimulating hormone (R = 0.47, p < 0.03); however, such a correlation was not observed in patients with nodular goiter. In conclusion, our findings suggest that the changes in the expression of regulatory proteins of the Bcl-2 family in the thyroid follicular cells indicate the involvement of apoptosis in the pathogenesis of GD.

Identification of apoptotic proteins in thyroid gland from patients with Graves' disease and Hashimoto's thyroiditis.

Apoptosis, i.e. natural programmed cell death, is a physiological phenomenon indispensable for normal functioning of the organism. The signal to apoptosis can be started practically in any cell. Disturbances in the apoptosis regulation determine the essential link of the pathogenesis of many diseases, including autoimmune thyroid disorders. The aim of the study was to assess the expression of Fas/FasL and caspase eight in the tissues of the thyroid gland in patients with Graves' disease (GD), non-toxic nodular goiter (NTNG) and Hashimoto's thyroiditis (HT). The analysis of Fas/FasL expression was performed by western blot and immunohistochemical investigation with DAB-visualization and Mayer's hematoxylin staining. Caspase-8 expression in thyroid follicular cells was assayed by western blot method. Identification of the proapoptotic proteins FasL and Fas exhibited their pronounced expression in the thyroid tissue in GD patients (++; ++) and HT (+++; +++) as compared to the NTNG group (0/+; 0/+). Among the study groups, the expression of caspase-8 was revealed in band 55 kDa from patients with autoimmune thyroid diseases. In GD patients, the percentage of thyrocytes with FasL expression correlated positively with TRAb (R = 0.58, p < 0.02). However, no such correlations were noted in HT or non-toxic multinodular goiter. There were no significant correlations between thyroid hormones and the percentage of thyrocytes with Fas and FasL expression. In conclusion, our findings suggest that the changes in the expression of apoptotic molecules on the surface of T lymphocytes and thyroid follicular cells in patients with autoimmune thyroid disorders reflect their substantial involvement in the pathogenesis of GD and HT. In addition, analysis of Fas/FasL and caspase-8 expression in thyroid tissue may indicate the disease activity and immunological phenotype.

Clinical and endocrine features and long-term outcome of Graves' disease in early childhood.

Hyperthyroidism is rare in early childhood and most commonly caused by Graves' disease. We report 14 children (4 boys, 10 girls) aged 3.4-7.5 yr. At diagnosis, all patients had weight loss, hyperkinetic activity, tachycardia, difficulty sleeping, and poor concentration and 11 presented with proptosis. Four patients developed long-term neuropsychological problems. There was a family history in 7 cases. All patients had goiters, clinically assessed to be large and diffuse in 21%, medium-sized in 43%, and small in 36%. At diagnosis, height was increased with median (range) height; 1.25 standard deviation score (SDS) (-0.2-5.24) and body mass index (BMI) was decreased; -0.48 SDS (-1.65-1.26). Height and BMI SDS values were statistically different (p<0.032) Bone age was advanced in 4 of 5 children, who had assessments. Total or free T4 levels were elevated and TSH was undetectable. Ninety percent of patients (12/14) had positive thyroid peroxidase autoantibodies, mean level 680 IU/ml (range 50-1347). Initial treatment was with antithyroid medication using carbimazole; median dose 0.75 mg/kg/day (no.=13) or propylthiouracyl 15 mg/kg/day (no.=1). T4 was added in 6 patients. Normalisation of serum T4 occurred at 4 months (1- 9) and TSH at 7 months (3-24) after start of therapy. Treatment was discontinued after a minimum of 2 yr in 11 patients, relapse occurring in 9. Median duration of total therapy was 58 months (18-132). During adolescence, 4 patients had curative therapy by surgery (no.=2) or radioiodine (no.=2). In conclusion, disturbance of growth, behavioral difficulties and infrequent spontaneous remission are key features of Graves' disease in early childhood.

Application of mouse monoclonal antibodies for identification of antigen regions of human thyroid peroxidase in adolescents with Graves' disease and non-toxic multinodular goiter by flow cytometry.

Thyroid peroxidase (TPO) is the major thyroid autoantigen recognized by serum autoantibodies from patients with Graves' disease (GD) or Hashimoto's thyroiditis directed to two immunodominant conformational regions termed A and B. The epitopes of human TPO have been defined using a panel of mouse monoclonal antibodies (mAbs). The aim of this study was to estimate the expression of chosen surface antigen regions of TPO (1, 18, 30, 64 epitopes) on thyroid cells in 15 patients with non-toxic multinodular goiter (NTMG) and 15 patients with GD. The thyrocytes were identified by indirect method: in the first stage we added mouse monoclonal autoantibodies specific for TPO regions and in the second stage we conjugated this complex with rabbit anti-mouse antibodies IgG (Fab')(2) with FITC. All investigations were performed by flow cytometry using Coulter EPICS XL apparatus. The percentages of thyrocytes with expression of epitopes 1, 18, 30, 64 TPO were measured in relation to the respective anti-TPO concentrations: 50-1600 microg/ml. The analysis of epitopes located in immunodominant regions (IDR) of TPO revealed higher percentages of thyrocytes in cases with GD in comparison to NTNG. The most predominant difference was observed for mAb 64 epitope (48 vs 7%, p < 0.019; 39 vs 5%, p < 0.017) at the concentration of 100-200 microg/ml mAbs. The expression of 18 epitope on thyrocytes was also statistically higher in Graves' patients than in the NTMG (14 vs 6%, p < 0.025) at concentration of 400 microg/ml mAbs. However, this expression was much less pronounced. In all the cases, the percentages of thyrocytes with epitopes 1 and 30 were in low detection (8-15% of positive cells). In conclusion, our findings suggest that the elevated expression of TPO epitopes 18 and 64 in young patients with thyroid autoimmune diseases increase stimulation and activation of thyroid cells during inflammatory reaction within the thyroid gland. In addition, predominant expression of 64 TPO epitope that recognizes B domain in GD patients could be a useful marker of the immune process in the thyroid gland.

Serum levels of cytokines in children and adolescents with Graves' disease and non-toxic nodular goiter.

It has been shown that human thyrocytes can synthesize cytokines which activate T and B lymphocytes. These immune cells play important roles in the initiation and continuation of thyroid autoimmunity. The aim of this study was to estimate serum concentrations of soluble interleukin-6 receptor (sIL-6R), interleukin-6 (IL-6) and interleukin-8 (IL-8) in patients with Graves' disease (GD) (n=44, mean age 14.8 years), in patients with nontoxic nodular goiter (NTNG) (n=36, mean age 15.6 years) and in a group of healthy controls (n=20, mean age 14.5 years). ELISA was used to determine the concentration of cytokines, antithyroglobulin and antithyroid peroxidase antibodies in patients with thyroid disease. Radio receptor assay (RRA) was performed to detect anti-TSH receptor autoantibodies (TRAb). Serum levels of IL-6, sIL-6R and IL-8 were markedly elevated in patients with GD before treatment with methimazole (p<0.0001 for IL-6, p<0.006 for sIL-6R, p<0.004 for IL-8) and after 8 weeks of therapy (p<0.011 for IL-6, p<0.04 for IL-8). However, following 24 months of treatment, normal serum concentrations of these cytokines were restored. Furthermore, patients with NTNG showed a slightly elevated concentration of cytokines (IL-6, IL-8). Serum levels of tri-iodothyronine in patients with GD positively correlated with serum concentrations of IL-6 (r = 0.35, p<0.025) and sIL-6R (r = 0.31, p<0.047), while no correlation was found between thyroxine and cytokines. Moreover, we observed a positive correlation between serum levels of TPO-Abs, TRAb and IL-6 (r = 0.43, p<0.008; r = 0.5, p<0.003) and between TPO-Abs and IL-8 (r = 0.67, p<0.0001). However, in patients with NTNG no correlation was observed between serum levels of antithyroid antibodies or thyroid hormones and serum levels of cytokines. We conclude that the cytokines (IL-6, sIL-6R, IL-8) could play an important role in the development of Graves' disease and that their levels are modulated by thyreostatic treatment.

Serum levels of adhesion molecules in children and adolescents with immune and non-immune thyroid diseases.

Serum levels of sICAM-1, sVCAM-1 and sP-selectin were determined in patients with Graves' disease before and after 8 weeks and 24 months of methimazole therapy, in levothyroxine suppressed patients with non-toxic nodular goiter, and in a group of healthy controls, to elucidate the relationship to circulating antithyroid antibodies and possible role of soluble adhesion molecules as markers of inflammatory activity. sICAM-1, sVCAM-1, and sP-selectin in patients with untreated hyperthyroidism were markedly elevated compared with controls. After 8 weeks of methimazole treatment, the concentrations of these molecules dropped, but were still significantly higher than in healthy children. In patients with clinical and biochemical remission after 24 months of therapy, sICAM-1 values were on the verge of significance, but still higher than in the control group, whereas serum levels of sVCAM-1 and sP-selectin had normalized. Slightly higher serum sICAM-1 values were observed in patients with non-toxic nodular goiter compared with healthy children.

[Serum levels of adhesion molecules (sICAM-1, sVCAM-1 and sP-selectin) in children and adolescents with thyroid diseases]. / Ocena stezenia wybranych czasteczek adhezyjnych (sICAM-1, sVCAM-1 i sP-selectin) u dzieci i mlodziezy w chorobach gruczolu tarczowego.

The targeting and recruitment of inflammatory cells to vascular endothelium in Graves' disease (GD) is mediated by intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and P-selectin. The aim of this study was to estimate the serum concentrations of soluble (sICAM-1), soluble (sVCAM-1) and soluble (sP-selectin) in patients with (GD) (n=44, mean age 14.8 years), with nontoxic nodular goiter (NTNG) (n=36, mean age 15.6 years) and in a group of healthy controls (n=20, mean age 14.5 years). ELISA method was used to determine the concentrations of adhesion molecules and antithyroglobulin and antithyroid peroxidase antibodies in patients with thyroid diseases. The anti-TSH receptor autoantibodies (TRAbs) were evaluated using a radio receptor assay (RRA). The serum levels of sICAM-1,sVCAM-1 and sP-selectin were markedly elevated in patients with GD before treatment with methimazole (p<0.0001 for all) and after 8 weeks of therapy (p<0.001 for sICAM-1 and sVCAM-1, p<0.014 for sP-selectin). After 24 months of therapy, serum concentrations of sVCAM-1 and sP-selectin were normalized, whereas serum level of sICAM-1 remained elevated. Furthermore, patients with NTNG showed a slighty elevated sICAM concentration. Serum levels of sICAM-1,sVCAM-1 and sP-selectin in patients with GD correlated with the serum concentrations of triiodothyronine (r=0.42, 0.45; 0.42, respectively) and thyroxine (NS, r=0.39; 0.46, respectively). Moreover, we observed a positive correlation between serum levels of TPO-Abs, TG-Abs, TRAbs and sICAM-1 (r=0.51; 0.40; 0.41 respectively) and sVCAM-1 (r=0.49; 0.38; 0.52, respectively). However, in patients with NTNG no correlation between serum level of adhesion molecules or thyroid hormones and serum level of antithyroid antibodies, respectively, was found. We conclude that the antithyroid antibodies involved in the pathogenesis of Graves' disease cause the elevation of the levels of adhesion molecules, which act as mediators of lymphocyte inflow and adhesion to the tissue of the thyroid gland. The evaluation of the serum level of soluble forms of adhesion molecules in children and adolescents with hyperthyroidism allows control over the autoimmune process.

[Cytokins and adhesion molecules in Graves's disease]. / Udzial wybranych cytokin i czastek adhezyjnych w chorobie Gravesa-Basedowa.

The course of thyroid autoimmune diseases is associated with the inflow of lymphocytes to the thyroid gland and with the expression of anti-TSH receptor, anti-thyroglobulin and antithyroid peroxidase antibodies. Vascular endothelium of the thyroid, which is not only the source of cytokines but also a location for adhesion molecules, plays a key role in the immune process. These microscopic glycoproteins participate in cell adhesion and transmission of intercellular information, which eventually leads to their activation or proliferation. Strongly expressed molecules within thyroid endothelium are involved in lymphocyte homing in the autoimmune process. A special role is assigned to ICAM and VCAM, which interact with lymphocytes or rather with integrins LFA-1 and VLA-4 present on their surface and responsible for adhesion and inflow of these cells to the sites of inflammation. The increased expression of adhesion molecules on endothelium is caused by the activation of locally produced cytokines (TNF-alpha, INF-gamma, IL-1, IL-6, sIL-6R), which are released during the inflammatory process. Such induction promotes migration of cells through endothelium during inflammation and contributes to the phasic inflow of various populations of leucocytes.

[Serum levels of interleukin 6 (IL-6) and soluble IL-6 receptor (sIL-6R) in children and adolescents with Graves-Basedow disease]. / Ocena stezenia interleukiny 6 (IL-6) i jej receptora (sIL-6R) u dzieci i mlodziezy w przebiegu choroby Gravesa-Basedowa.

The concentration of interleukin 6 (IL-6), soluble IL-6 receptor (sIL-6R) and intercellular adhesion molecule-1 (sICAM-1) was assayed in 24 children with Graves' disease, aged 6.5-17 years, before treatment with methimazole, during clinical and biochemical remission (8 weeks), and following the eighteen-month therapy with the thyrostatic. ELISA method was used to determine the concentration of interleukin 6, sIL-6R and sICAM-1. An increase was observed in serum concentration of IL-6 and its receptor sIL-6R and soluble intercellular adhesion molecule-1 in the course of Graves' disease. A significant reduction in sIL-6R (8 weeks) and IL-6 (18 months) was noted after therapy with methimazole. A positive correlation was found between microsomal antibodies and the concentration of sICAM-1 and sIL-6R, and between antithyroglobulin antibodies and concentration of sIL-6R in children with fresh hyperthyroidism. No correlation was revealed between the level of thyroid hormones and the concentration of cytokines and adhesion molecules.