concanavalin-a and Thyroid-Diseases

The composition of sugar chains on thyroglobulin (Tg) produced in thyroid carcinoma cells (C-Tg) is different from Tg produced in normal thyroid tissues (N-Tg). In this study, we designed a new method for detecting Tg derived from thyroid carcinoma based on the differences between C-Tg and N-Tg in the reactivity with lectins.. Thyroglobulin preparations obtained from various thyroid tissues were incubated with lectins, and the amount of lectin-unbound Tg (ub-Tg) in the supernatant relative to Tg untreated with lectin was determined by enzyme-linked immunosorbent assay and expressed as ub-Tg(%). In addition, to study further the differences in glycosylation between C-Tg and N-Tg, concanavalin A binding to Tg digested with Staphylococcus aureus V8 protease was analyzed on nitrocellulose membrane after Western blotting.. The ub-Tg(%) in C-Tg from papillary carcinoma was significantly higher than in Tg from Graves' disease, benign goiter, and normal thyroid tissue for both concanavalin A and ricinus communis agglutinin-120. Concanavalin A did not appear to bind to Tg from papillary carcinoma after V8 treatment by Western blot analysis. The ub-Tg(%) in Tg from follicular adenoma was significantly higher than C-Tg from follicular carcinoma, whereas there were no differences in ub-Tg(%) between follicular carcinoma and normal thyroid tissue in concanavalin A treatment.. These results suggest our new methods can distinguish both between C-Tg from papillary carcinoma and N-Tg, and between follicular carcinoma and follicular adenoma in thyroid tissue specimens. Thus, this type of analysis may be applicable to differentiate C-Tg from N-Tg in thyroid aspirates for the adjunctive cytodiagnosis of thyroid carcinoma.

Topics: Biomarkers; Blotting, Western; Concanavalin A; Histocytochemistry; Humans; Lectins; Plant Lectins; Sensitivity and Specificity; Thyroglobulin; Thyroid Diseases; Thyroid Gland; Thyroid Neoplasms; Wheat Germ Agglutinins

Patients with nonthyroid illness (NTI) often have reduced serum T3, free T3, T4, and free T4 concentrations. Paradoxically, serum TSH is usually in the normal range. The data suggest a diagnosis of hypothalamic hypothyroidism, in which TSH may have reduced biological activity because TRH, which is necessary for key steps in the glycosylation of TSH, is deficient. To study the glycosylation of TSH in patients with NTI, we measured the serum TSH concentration in 36 such patients hospitalized on our intensive care units and compared the results with those from a group of 18 normal subjects. Serum TSH was measured in 2 assays: 1) a sensitive TSH RIA of unextracted serum (TSH-RIA) and 2) a RIA of serum TSH after its extraction with Concanavalin-A (Con-A), a lectin which binds glycoproteins containing mannose residues in their oligosaccharide side-chains (TSH-Con-A). The ratio of TSH-Con-A to TSH-RIA was significantly reduced in the NTI patients [0.61 +/- 0.03 (+/- SE) vs. 0.89 +/- 0.05 in the normal subjects] due to reduced binding of the TSH to the Con-A. This change was not dependent on the extent of the abnormalities of thyroid hormone levels. The data suggest that the TSH secreted in NTI has altered glycosylation which is associated with reduced biological activity. This finding may explain in part the low serum T4 level in NTI patients in the face of an apparently normal immunoreactive TSH level.

Topics: Adult; Aged; Concanavalin A; Heart Diseases; Humans; Lung Diseases; Middle Aged; Protein Binding; Radioimmunoassay; Thyroid Diseases; Thyroid Hormones; Thyrotropin; Thyroxine

We examined the expression of HLA-DR antigen induced by mitogen, mitogen-free supernatants from mitogen-stimulated peripheral blood mononuclear cells (PBMC), or autologous and allogeneic PBMC on thyrocytes cultured for 1-2 weeks (precultured) before the addition of the stimulant. Leucoagglutinin (LAG) and concanavalin A, but not lipopolysaccharide induced HLA-DR expression on thyrocytes from normal subjects (NC) and patients with Graves' disease (GD) and Hashimoto's thyroiditis (HT). The degree of DR expression induced by LAG was significantly less in GD than in NC thyrocytes. This response was dependent on contaminating T cells, especially suppressor-cytotoxic T (Ts/c) cells, NK cells, and HLA-DR+ cells, but not helper-inducer T (Th/i) cells or B cells, in the thyrocyte cultures. OKT3 monoclonal antibody, which activates T cells specifically in the presence of monocytes, also induced thyrocyte HLA-DR expression. Furthermore, interferon-gamma (IFN-gamma) was detected in culture supernatants from LAG-stimulated thyrocytes. Anti-IFN-gamma monoclonal antibody eliminated the ability of LAG to induce HLA-DR. Mitogen-free supernatants from mitogen-stimulated PBMC also induced thyrocyte HLA-DR expression, which was inhibited by anti-IFN-gamma. The supernatants of concanavalin A- or LAG-stimulated PBMC from either untreated or recently treated patients with GD or hypothyroid HT induced less thyrocyte DR expression than NC PBMC. Indeed, the levels of IFN-gamma in supernatants from such patients were lower than those in NC, and the correlation between DR expression and IFN-gamma levels was significant. This IFN-gamma production by PBMC required Th/i cells, NK cells, and HLA-DR+ cells. Before the addition of autologous or allogeneic PBMC, only precultured HT thyrocytes expressed HLA-DR, whereas GD and NC thyrocytes did not. The induction or enhancement of DR expression on autologous thyrocytes by direct coculture with PBMC occurred within 8 days in GD and HT, but not in NC. There was a significant correlation between the serum titer of antithyroid microsomal antibodies and the degree of DR expression. Allogeneic normal PBMC also induced DR expression on NC and GD thyrocytes within 8 days, the effect on the latter being more pronounced than with autologous GD PBMC. Thyrocyte HLA-DR expression induced by autologous GD PBMC and allogeneic normal PBMC required monocytes. Th/i, and NK cells and was blocked by anti-IFN-gamma. However, the enhancement of thyrocyte DR expre

Topics: Adult; Aged; Agglutinins; Antibodies, Monoclonal; Autoimmune Diseases; Cells, Cultured; Concanavalin A; Fluorescent Antibody Technique; Histocompatibility Antigens Class II; HLA-DR Antigens; Humans; Interferon-gamma; Leukocytes; Lipopolysaccharides; Middle Aged; Proteins; T-Lymphocytes; Thyroid Diseases; Thyroid Gland

Con A-induced production of interleukin 2 was studied in peripheral blood lymphocytes obtained from 15 normal subjects and 42 patients with different thyroid diseases. It was found that thyroid diseases are accompanied by changes in IL 2 production. The level of IL 2 production by T lymphocytes is determined by the functional status of thyroid glands, their anatomy, stages of the disease and mode of treatment.

Topics: Adolescent; Adult; Aged; Concanavalin A; Humans; Interleukin-2; Lymphocyte Activation; Middle Aged; T-Lymphocytes; Thyroid Diseases

High ferritin levels in the aspirate of thyroid cyst (Six yellow clear, 4 yellow turbid and 10 chocolate colored turbid) without apparent sings of malignancy were found. The mean concentration in the 3,000 X g supernatant of the fluid was 40,116 ng/ml, and the 3,000 X g precipitate was 11,147 ng/ml. All cases showed normal levels of serum ferritin. Con A binding with ferritin was distributed from low to high. These ferritins showed a molecular weight of approximately 450,000 which was the same as found in human spleen and liver. The continuous increase in ferritin levels in thyroid cyst fluid was found by a chronological study in some cases. When tumor markers such as CEA, NCA, CA 19-9 and alpha 1-acid glycoprotein (alpha 1-AG, acute phase reactant) were examined simultaneously, an increase in some of the cyst fluid was observed. However, the incidence and the rate of increase of these tumor markers and acute phase reactant were low compared to ferritin. Neither a correlation between ferritin and CEA, nor between ferritin and CA 19-9 was found. The increase in ferritin in thyroid cyst fluids may be due to the increased synthesis, release and storage by the inflammatory cells.

Topics: Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carcinoembryonic Antigen; Chromatography, Gel; Concanavalin A; Cysts; Exudates and Transudates; Female; Ferritins; Humans; Male; Molecular Weight; Thyroglobulin; Thyroid Diseases; Thyroid Function Tests

Human thyroglobulin (Tg) immune complexes (TgIC) were preformed at different ratios from purified Tg, and Tg antibodies (TgAb) obtained from six patients with autoimmune thyroid diseases. TgIC were characterized by precipitation with polyethylene glycol, and sedimentation in sucrose gradient. TgIC from one patient were further characterized by concanavalin A (con A), and binding to the complement factor Clq and rheumatoid factor (RF). Tg and TgIC had almost identical binding to con A, no binding to Clq, but could be partially separated by ultracentrifugation and polyethylene glycol. The highest degree of separation was obtained by a RF-coated plastic tube radiometric assay, using 125-I-rabbit TgAb as indicator. Tg showed no binding to RF, and a dose-response curve of TgIC in serum could be established. There was a dependence of the Tg-TgAb ratio, TgIC at equilibrium and in antibody excess being detected most efficiently. The method may serve as an aid in the evaluation of the fate of Tg, TgAb and TgIC following thyroid damage (surgery, radioiodine) and may be extended as a model system in the investigation of immune complexes in connection with autoimmune disorders.

Topics: Antigen-Antibody Complex; Antigens; Autoantibodies; Autoantigens; Complement C1; Concanavalin A; Humans; Models, Biological; Radioisotopes; Rheumatoid Factor; Thyroglobulin; Thyroid Diseases; Ultracentrifugation

Topics: Adolescent; Adrenal Gland Diseases; Animals; Antibody Formation; Antibody-Producing Cells; Autoimmune Diseases; B-Lymphocytes; Concanavalin A; Cytotoxicity Tests, Immunologic; Female; Genital Diseases, Female; Gonads; Humans; Immunity; Immunoglobulin Fc Fragments; Immunoglobulin G; Lectins; Lymphocytes; Mitogens; Sheep; T-Lymphocytes; Thyroid Diseases