Interaction of lymphocytes and thyrocytes in Graves' disease and nonautoimmune thyroid diseases in immunohistochemical and ultrastructural investigations.

BACKGROUND: Graves' disease is the archetype for organ-specific autoimmune disorders. It is very important for our understanding of the mechanisms responsible for progression of autoimmunity. The aim of this study was to present interactions of lymphocytes and thyrocytes in the thyroid tissue in Graves' disease and nonautoimmune thyroid diseases. METHODS: The study involved 30 children with Graves' disease, 30 children with nodular goiter, 30 with simple goiter and 30 healthy children. After thyroidectomy, T cells were detected in the thyroid specimens by CD3, CD4, CD8 antibodies, B cells by CD79alpha antibodies and the antigen-presenting dendritic cells with CD1a antibodies (DakoCytomation) and were examined in the EM 900 Zeiss Germany Electron Microscope. RESULTS: The most enhanced immune reaction was observed in the thyroid from children with Graves' disease. The cells of the immune system infiltrated the thyroid follicles and interfollicular compartments; they also formed lymph follicles. CONCLUSION: The immune reaction in Graves' disease and migration of lymphocytes T and B between thyrocytes results in the thickening of the basal membrane of the thyroid follicle. No cytotoxic effect of T cytotoxic/suppressor CD8+ cells on thyrocytes was observed in Graves' disease, while a mild cytotoxic effect was observed in non-autoimmune thyroid disease.

Treatment of severe primary IGF-1 deficiency using rhIGF-1 preparation - first three years of Polish experience.

INTRODUCTION: The objective of this study was to analyse the effects of the first three years of treatment with recombinant human insulinlike growth factor 1 (rhIGF-1) in patients from the Polish population. MATERIAL AND METHODS: Twenty-seven children (22 boys and five girls) aged 2.8 to 16.0 years old were qualified for treatment with rhIGF-1 (mecasermin) in different treatment centres, according to Polish criteria: body height below -3.0 SD and IGF-1 concentration below percentile 2.5 with normal growth hormone (GH) levels. Mecasermin initial dose was 40 µg/kg bw twice a day and was subsequently increased to an average of 100 µg/kg bw twice a day. Body height, height velocity, weight, body mass index (BMI), and adverse events were measured. RESULTS: Mecasermin treatment resulted in a statistically significant increase in body height (1.45 ± 1.06 SD; p < 0.01) and height velocity in comparison with pre-treatment values. The biggest change in height velocity happened during the first year and diminished during subsequent years. Body weight and BMI also increased significantly after treatment (1.16 ± 0.76 SD and 0.86 ± 0.75 SD, respectively; p < 0.01). Eight patients reported adverse events. These were mild and temporary and did not require treatment modification except in two patients. CONCLUSIONS: Treatment with rhIGF-1 was effective and safe in Polish patients with primary IGF-1 deficiency. It had a clear beneficial effect on the height of the patients and significantly accelerated the height velocity, particularly in the first year of treatment.

[Vitamin D3 overdosage due to rashly diagnosed rachitis in a child with distal tubular acidosis]. / Przedawkowanie witaminy D3 z powodu pochopnego rozpoznania krzywicy u dziecka z kwasica cewkowa dystalna.

Metabolic acidoses are diseases causing many diagnostic and therapeutic problems. Compensated metabolic acidosis can be unrecognised for a long time. This refers especially to isolated renal tubular acidosis (RTA). Unrecognised RTA causes calcium and phosphorus balance disturbances with clinical signs of improper bone mineralization. It happens that some patients with mentioned problems are "treated" as rachitic and take high doses of vitamin D. As a result, serum calcium and phosphates as well as urine calcium increase, without the satisfied influence on bone mineralization. We present a case of a 3.5 months old baby, who was "treated" as ricket in vitamin D deficiency. This baby was "cured" with high doses of cholecalciferol (0.0875 mg/24h for 2 weeks, then 0.175 mg/24h for 3 weeks) because of craniotabes. This treatment was carried on without any metabolic tests and caused the following disturbances: 25(OH)D serum level - 102.7 ng/ml (normal 11-54), 1,25(OH)2D serum level - 39.5 pg/ml (normal 15-70), calcaemia 2.7-2.85 mmol/l, phosphataemia 2.1 mmol/l. In this time the considerable hipercalciuria (second morning urine sample Ca/cr ratio 2.06 mmol/mmol) occurred. The other laboratory test showed as follows: serum albumins 4.5 g/dl, alkaline phosphatase 188 U/l, acid phosphatase 10.7 U l, Cl- 114.9 mmol/l, Na 146 mmol/l, K 4.8 mmol/l, HCO3a 16.6-20.1 mmol/l and pCO2 3.63-3.85 kPa, serum anion gap 11 mEq/l; pH of morning urine 6.5-7. These results suggested the presence of distal RTA aside from symptoms of vitamin D overdosage. The high serum levels of calcium and phosphates, craniotabes, rather low serum alkaline phosphatase activity and presence of metabolic acidosis the symptoms after the normalisation of calcium and phosphorus balance suggested that the distal RTA had been prior to calcium disturbances.

[Evaluation of lipids, lipoproteins and apolipoproteins in the blood serum of children with type 1 diabetes]. / Zachowanie sie stezen lipidów, lipoprotein i apolipoprotein w surowicy krwi u dzieci z cukrzyca typu 1.

BACKGROUND: Children and youth with type 1 diabetes have a higher risk of atherosclerosis. Therefore all of the atherosclerosis risk factors should be observed in those children early. The disorders of lipid metabolism are characteristic and their intensification depends on the type of diabetes and its balance. OBJECTIVES: The objective of the paper was to evaluate behaviour of lipids, lipoproteins and apolipoproteins behaviour in the blood serum of children with type 1 diabetes. MATERIAL AND METHODS: The study included 53 children (23 girls and 30 boys) with type 1 diabetes aged 8-16 years suffering from diabetes for 3-10 years. During the study the children were in the state of relative metabolic balance. In all of the studied subjects concentration of triglycerides, total and HDL cholesterol, apolipoproteins (Apo-AI, Apo-B) were assayed in the blood serum. The level of LDL and VLDL cholesterol was determined with an indirect method. The obtained results were statistically analysed and compared with the earlier developed own standards for the healthy children. RESULTS: In the girls with type 1 diabetes higher levels of triglycerides and of total cholesterol, LDL and VLDL cholesterol as well as of Apo-B were confirmed and a lower level of HDL cholesterol was noticed as compared with the control group. But in the boys with type 1 diabetes higher level of LDL cholesterol and lower level of HDL cholesterol were confirmed as compared with the healthy children. CONCLUSIONS: 1) In the children and youth with type 1 diabetes the disorders of lipid metabolism were confirmed. 2) Periodical evaluation of lipid metabolism should be taken into account in monitoring children with type 1 diabetes.

Treatment of Graves' disease with methimazole in children alters the proliferation of Treg cells and CD3+ T lymphocytes.

Almost all cases of hyperthyroidism in children result from Graves' disease (GD). Recent studies have confirmed a significant role of T regulatory cells (Tregs) in the development of autoimmune diseases. However, the interactions between T cell responses and Treg proliferation in GD are still poorly understood. The aim of this study was to assess the proliferation of Treg cells (Tregs) and CD3+ T lymphocytes isolated from 50 children with GD before and after treatment with the thyreostatic drug methimazole (MMI). The proliferation rates, measured by methyl-3H-thymidyne incorporation, of CD3+ cells and Tregs stimulated with mitogen phorbol 12-myristate 13-acetate (PMA) were compared with those of unstimulated cells. The proliferation rates of both PMA-stimulated and unstimulated CD3+ cells prior to treatment with MMI were significantly higher than after treatment. Simultaneously, the proliferation rates of both PMA-stimulated and unstimulated Tregs were significantly lower before MMI treatment. Moreover, we observed higher cell proliferation rates of unstimulated and PMA-stimulated Tregs before the initiation of MMI therapy and after treatment in patients who had no relapse of hyperthyroidism. There was a positive correlation between the CD3+ cells proliferation rate before MMI treatment and fT3, as well as fT4 concentration in peripheral blood. The proliferation rates of CD3+ T cells before and after MMI treatment positively correlated with the TSI index. Thus, children suffering from Graves' disease presented lower Tregs proliferative potential compared with CD3+ T cells. Cocultures of CD3+ T cells and Tregs showed that Tregs were not capable of efficiently inhibiting the proliferation of CD3+ T cells in GD patients. Conclusions. MMI treatment reduced the proliferative activity of CD3+ T cells in pediatric GD patients and increased the proliferation rate of Tregs. We suggest that Treg cells that are partly dysfunctional in GD disease are probably suppressed by CD3+ T cells and that methimazole exerts some immunomodulatory effects.

The differences in T and B cell subsets in thyroid of children with Graves' disease and Hashimoto's thyroiditis.

BACKGROUND: The differences between Graves' disease (GD) and Hashimoto's thyroiditis (HT) suggest that changes in the subsets of T cells may have an influence on the course of these reactions. METHODS: This study included 90 children: 30 with GD, 30 with HT, and 30 healthy children as controls. After thyroidectomy, standard histological examinations and immunohistochemical reactions were performed in paraffin specimens with monoclonal antibodies against T cell markers CD3, CD4, CD8 as well as against CD79 alpha B cells. Ultrathin sections were examined under a transmission electron microscope. RESULTS: Autoimmune reaction in GD consisted of an increased number of CD4+ T cells (3.17±4.27%) and plasma cells (22.89±8.61%) producing thyroidstimulating hormone-receptors and stimulating thyrocytes to activity. The number of CD8+ T cells was increased in children with HT (20.54±0.68%) as compared with the controls (0.65±0.30%). The autoimmune reaction in the HT children showed antibody dependent cytotoxicity with a low number of CD4+ T cells and an increased number of CD8+ T cells in the thyroid tissue in comparison with that in the GD children and the controls. Plasma cells (31.65±9.11%) in this situation produced the antibodies involved in cytotoxic reactions against thyrocytes. CONCLUSIONS: Graves' disease is characterized by the increased number of CD4+ T cells and CD8+ T cells. Hashimoto's thyroiditis is characterized by the low number of CD4+ T cells and increased number of CD8+ T cells. CD8+ T cells have cytotoxic properties only in Hashimoto's thyroiditis.

Interactions of lymphocytes, thyrocytes and fibroblasts in Hashimoto's thyroiditis: an immunohistochemical and ultrastructural study.

BACKGROUND: The mechanism of autoimmune reaction, a diffuse process consisting of a combination of epithelial cell destruction, lymphoid cellular infiltration, and fibrosis in Hashimoto's thyroiditis, is not well known. The aim of this study was to analyse the cell subsets in thyroid tissue of patients with Hashimoto's thyroiditis. METHODS: We studied paraffin-embedded thyroid specimens obtained from children with Hashimoto's thyroiditis and children without an autoimmune thyroid disease. Mononuclear T cells were detected by means of CD3+, CD4+, CD8+ antibodies, B cells by CD79 alpha+ antibodies, and antigen-presenting cells by CD1a+ antibodies, and they were counted in every 1,000 cells. The specimens from each patient were routinely estimated and investigated under the electron microscope. RESULTS: In Hashimoto's thyroiditis, we observed a statistically significant increase in T suppressor/cytotoxic cells CD8+ (20.54 ± 0.68%) in comparison to the control group (0.65 ± 0.30%), simple goitre (4.01 ± 5.54%) and nodular goitre (8.53 ± 2.37%), and a statistically significant increase in plasma CD79 alpha+ cells (31.65 ± 9.11%) in comparison to the control group (4.11 ± 1.94%), simple goitre (1.83 ± 0.64%) and nodular goitre (5.22 ± 1.63%). Simultaneously, we observed a low number of CD4+ T helper cells in the thyroid gland (0.93 ± 0.99%) in Hashimoto's thyroiditis (0.19 ± 0.05% in the control group, 1.05 ± 2.71% in simple goitre, 2.03 ± 1.06% in nodular goitre). The ultrastructural investigations showed interactions between T cells, plasmocytes, fibrocytes and thyrocytes leading to apoptosis of thyrocytes. An immunological synapse between T cells, plasmocytes and thyrocytes in the thyroid gland was noticed. CONCLUSIONS: In Hashimoto's thyroiditis, autoantigen presentation in combination with a low number of CD4+ T helper cells and a high number of CD8+ cells and plasmocytes caused the development of a cytotoxic reaction against thyrocytes, leading to apoptosis of the thyrocytes.

[A patient with Klinefelter's syndrome and growth hormone deficiency]. / Pacjent z zespolem Klinefeltera i niedoborem hormonu wzrostu.

INTRODUCTION: Short stature in Klinefelter's syndrome was reported sporadically and was connected with the karyotype 49,XXXXY. AIM OF THE STUDY: The presentation of growth in a boy with Klinefelter's syndrome and somatotropine deficiency. CASE REPORT: A 10-year-old boy was examined in the Department of Endocrinology because of short stature and mild mental retardation. On admission, a height 117 cm (<3 percentile) and a body weight 20 kg (<3 percentile) were reported. The boy was in the prepubertal stage of life. The bone age was delayed by 5 years in comparison to the calendar age. Serum levels of thyroid hormones were within the normal range. Deficiency of growth hormone was diagnosed because maximal serum levels of growth hormone in stimulating tests with clonidine and insulin did not exceed 10.7 muIU/ml. During substitutive therapy with growth hormone an acceleration of growth rate was achieved. Because of the lack of consent to therapy from the boy and his family, the treatment was withheld after 1 year and 7 months - the boy was 135.6 cm high then. The patient was again examined in the Department of Endocrinology at the age of 14 years and 8 months because of breast enlargement. He had grown 25.4 cm and was 161.0 cm high (10 percentile). Puberty stage G1P3A3 was observed. Because of the low testosterone and increased gonadotropin serum levels hypergonadotropic hypogonadism was diagnosed and testosterone therapy was employed. The genetic investigation revealed Klinefelter's syndrome with 47,XXY karyotype. In the 16th year of life the patient was 175 cm high (25-50 percentile). CONCLUSION: Patients with Klinefelter's syndrome may accelerate growth rate in the pubertal period of life even if they have growth hormone deficiency.