scholarly journals Clinical Significance of a Sensitive Assay for Thyroid-Stimulating Antibodies in Graves' Disease Using Polyethylene Glycol at High Concentrations and Porcine Thyroid Cells.

1999 ◽  
Vol 46 (3) ◽  
pp. 397-403 ◽  
Author(s):  
KEIICHI KAMIJO ◽  
ATSUO NAGATA ◽  
YASUSHI SATO
Keyword(s):  
Polyethylene Glycol ◽  
Clinical Significance ◽  
Graves Disease ◽  
Sensitive Assay ◽  
Thyroid Cells ◽  
Thyroid Stimulating Antibodies ◽  
High Concentrations

A sensitive and practical assay for thyroid-stimulating antibodies using FRTL-5 thyroid cells

1987 ◽  
Vol 115 (1) ◽  
pp. 30-36 ◽  
Author(s):  
Kanji Kasagi ◽  
Junji Konishi ◽  
Yasuhiro Iida ◽  
Yasutaka Tokuda ◽  
Keisuke Arai ◽  
...  
Keyword(s):  
Fold Increase ◽  
Graves Disease ◽  
Thyroid Cells ◽  
Thyroid Stimulating Antibodies ◽  
Silent Thyroiditis ◽  
Poorly Differentiated ◽  
Pre Treatment ◽  
Rat Thyroid ◽  
Serum Tsh ◽  
Bovine Tsh

Abstract. A sensitive, precise and practical assay for thyroid stimulating antibodies was developed in which poorly differentiated rat thyroid cells (FRTL-5) were exposed to crude immunoglobulin fractions precipitated from serum with 15% polyethylene glycol under hypotonic conditions. After the incubation at 37°C for 2 h, cAMP released into Hank's medium without NaCl was determined by radioimmunoassay. The removal of NaCl from the isotonic Hank's medium greatly enhanced cAMP production in response to both TSH and thyroid stimulating antibodies. The assay was sensitive enough to elicit an approximately 30-fold increase in cAMP at 10 mU/l bovine TSH. Thyroid stimulating activities measured using FRTL-5 cells significantly correlated with those measured using cultured porcine (r = 0.918, N = 72) or human (r = 0.830, N = 23) thyroid cells. Thyroid stimulating activities were detected in all of the 50 patients with hyperthyroid Graves' disease, the 14 patients with recurrent hyperthyroid Graves' disease, and the 25 patients with ophthalmic Graves' disease. Thyroid stimulating activity was also detected in some patients (9/24, 37.5%) with Hashimoto's thyroiditis whose serum TSH concentrations were higher than 30 mU/l. However, it was completely abolished by pre-treatment of the sera with anti-TSH antibodies. Although thyroid stimulating activities were detected in one of the patients with simple goitre (N = 10) and in one with thyroid cancer (N = 10), none of the patients with silent thyroiditis (N = 7), adenomatous goitre (N = 11), and thyroid adenoma (N = 9) were positive for thyroid stimulating antibodies.

A sensitive method for assaying thyroid stimulating immunoglobulins of Graves' disease: use of the guanyl nucleotide-amplified thyroid adenylate cyclase assay

1983 ◽  
Vol 103 (3) ◽  
pp. 345-351 ◽  
Author(s):  
E. Macchia ◽  
P. Carayon ◽  
G. F. Fenzi ◽  
S. Lissitzky ◽  
A. Pinchera
Keyword(s):  
Adenylate Cyclase ◽  
Hashimoto’S Thyroiditis ◽  
Plasma Membranes ◽  
Hashimoto's Thyroiditis ◽  
Graves Disease ◽  
Tissue Preparation ◽  
Thyroid Cells ◽  
Significant Stimulation ◽  
Adenylate Cyclase Stimulation ◽  
Sensitive Method

Abstract. The purpose of this study was to develop and validate a sensitive method for evaluating adenylate cyclase stimulation by thyroid-stimulating antibodies (TSAb), based on the measurement of thyroid membrane adenylate cyclase activity in the presence of a non-hydrolyzable GTP analogue, guanyl-5'-yl imidodiphosphate (Gpp(NH)p). The addition of Gpp(NH)p (10−5 m) produced a 10-fold increase of the sensitivity of the system for both TSH and TSAb. Immunoglobulin G preparations from sera of 30 patients with Graves' disease were tested for the adenylate cyclase stimulation either in the presence or in the absence of Gpp(NH)p: a significant stimulation was observed in 27/30 patients when the GTP analogue was added to the system, while only 20/30 patients were positive in the absence of the nucleotide. The advantage of Gpp(NH)p addition was also evident in a large series which included 57 patients with Graves' disease, 15 with Hashimoto's thyroiditis or primary myxoedema and 22 normal subjects. In fact, 88% of patients with Graves' disease resulted positive, while no significant stimulation was elicited by Hashimoto's thyroiditis, primary myxoedema and by normal immunoglobulins. The sensitivity achieved in our system which employs thyroid plasma membranes was similar to that obtained by other investigators with the use of thyroid slices or thyroid cells in primary culture. Furthermore, methods based on thyroid plasma membranes are supposed to have a better reproducibility, since the same tissue preparation, if appropriately stored, may be used in several different tests.

Clinical significance of anti-TSH antibody in sera from patients with Graves’ disease and other thyroid disorders

1989 ◽  
Vol 12 (7) ◽  
pp. 483-488 ◽  
Author(s):  
T. Akamizu ◽  
Toru Mori ◽  
H. Imura ◽  
J. Noh ◽  
N. Hamada ◽  
...  
Keyword(s):  
Clinical Significance ◽  
Graves Disease ◽  
Thyroid Disorders

The effects of cytokines, antithyroidal drugs and glucocorticoids on phagocytosis by thyroid cells

1988 ◽  
Vol 119 (3) ◽  
pp. 413-419 ◽  
Author(s):  
Mayumi Matsunaga ◽  
Katsumi Eguchi ◽  
Takaaki Fukuda ◽  
Hiroshi Tezuka ◽  
Yukitaka Ueki ◽  
...  
Keyword(s):  
Phagocytic Activity ◽  
Interleukin 1 ◽  
Thyroid Tissue ◽  
Interferon Α ◽  
Graves Disease ◽  
Dependent Manner ◽  
Normal Thyroid Tissue ◽  
Thyroid Cells ◽  
Latex Beads ◽  
Thyroid Glands

Abstract. The present study was undertaken to examine whether thyrocytes possess phagocytic activity and whether the phagocytic activity is influenced by cytokines, such as interleukin 1, 2 (IL 1, IL 2) and interferon-α, -β, and -γ (IFN-α, β, and γ), and drugs, such as methimazole and dexamethasone. Thyroid glands were obtained from patients with Graves' disease. Thyrocytes were prepared by collagenase digestion. Thyrocytes were pre-incubated in the presence or absence of cytokines and drugs at 37°C for 20 h and were further incubated with fluoresceinated latex beads at 37°C for 60 min. The number of phagocytic thyrocytes was determined by FACS IV. Phagocytosis of latex beads was indeed seen within thyrocytes and gradually increased in a time-dependent manner. The rate of phagocytosis in thyrocytes was extremely slow as compared with that in macrophages. Phagocytic activity was detected in thyrocytes from patients with Graves' disease and from normal thyroid tissue adjacent to thyroid cancer. Phagocytosis was inhibited by IL 1, but was enhanced by IL 2. Although the enhanced phagocytosis with IFN-β was consistently seen, little effect was detected with IFN-α and -γ. Both methimazole and dexamethasone markedly inhibited phagocytosis. These results indicated that thyrocytes had phagocytic properties and that their phagocytic activity was modulated by cytokines, antithyroidal drugs and dexamethasone.

Studies of membrane fusion. III. Fusion of erythrocytes with polyethylene glycol

1979 ◽  
Vol 36 (1) ◽  
pp. 61-72
Author(s):  
S. Knutton
Keyword(s):  
Polyethylene Glycol ◽  
Cell Fusion ◽  
Close Contact ◽  
Freeze Fracture ◽  
Cell Swelling ◽  
Adjacent Cell ◽  
Cell Contact ◽  
Cell Agglutination ◽  
High Concentrations ◽  
Freeze Fracture Electron Microscopy

Freeze-fracture electron microscopy has been used to investigate the mechanism of polyethylene glycol-induced cell fusion. Interaction of cells with the high concentrations of polyethylene glycol required for cell fusion results in cell agglutination with large planar areas of very close contact between adjacent cell membranes. An aggregation of intramembrane particles into large patches at the sites of cell-cell contact accompanies cell agglutination. Fusion occurs following the removal of most of the PEG when cells only remain in close contact at small (approximately 0.1 micrometer diameter) plaques of smooth membrane resulting in cells connected by one (or more) small cytoplasmic connexions. Expansion to form spherical fused cells occurs by a process of cell swelling.

scholarly journals Low Doses of Methylmercury Induce the Proliferation of Thyroid Cells In Vitro Through Modulation of ERK Pathway

2020 ◽  
Vol 21 (5) ◽  
pp. 1556 ◽  
Author(s):  
Valentina Maggisano ◽  
Stefania Bulotta ◽  
Marilena Celano ◽  
Jessica Maiuolo ◽  
Saverio Massimo Lepore ◽  
...  
Keyword(s):  
Cell Viability ◽  
Prolonged Exposure ◽  
Signal Transduction Pathway ◽  
Immunoblot Analysis ◽  
Thyroid Cell ◽  
Promoting Effect ◽  
Thyroid Cells ◽  
M Phase ◽  
High Concentrations

Exposure to environmental endocrine disruptors has been associated with an increased frequency of thyroid pathology. In this study, we evaluated the effects of various concentrations of methylmercury (MeHg) on immortalized, non-tumorigenic thyroid cells (Nthy-ori-3-1). Exposure to MeHg at 2.5 and 5 µM for 24 h caused a reduction in cell viability with a decrease of the cell population in sub-G0 phase, as detected by MTT and flow cytometry. Conversely, MeHg at the lower concentration of 0.1 µM increased the cell viability with a rise of G2/M phase. An immunoblot analysis showed higher expression levels of phospho-ERK and not of phospho-Akt. Further enhancement of the cell growth rate was observed after a prolonged exposure of the cells up to 18 days to MeHg 0.1 µM. The present findings demonstrate the toxicity of high concentrations of MeHg on thyroid cells, while showing that treatment with lower doses of Hg, as may occur after prolonged exposure to this environmental contaminant, exerts a promoting effect on thyroid cell proliferation, by acting on the ERK-mediated pro-oncogenic signal transduction pathway.

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