Transient lack of response to TSH of human cultured thyroid cells obtained from hyperfunctioning tissue

1986 ◽  
Vol 113 (3) ◽  
pp. 346-354
Author(s):  
Annalisa Tanini ◽  
Maria Luisa Brandi ◽  
Umberto Modigliani ◽  
Carlo M. Rotella ◽  
Roberto Toccafondi

Abstract. TSH-induced cAMP accumulation in cells obtained from normal and pathological thyroid tissue was studied during the first 12 days of primary culture. In normal thyroid tissue cultures (N = 7), the response of cAMP to TSH was present from the second day of culture and reached its maximum after 8 days. A similar behaviour was observed in cultures obtained from euthyroid sporadic goitres (N = 8), even if the rate of response was slightly lower than that of normal tissue. Similarly, cultured cells from euthyroid 'autonomous' nodules (N = 8) appeared to be responsive to TSH during the period of study, but the rate of response was also lower than in the controls. On the contrary, in cultures obtained from toxic adenomas (N = 5) and from diffuse toxic goitres (N = 5) the response to TSH was absent during the first 4 days of culture. The cells became sensitive to TSH from 6 and 6 day onwards, with the rate of response increasing progressively and reaching its maximum on day 12. Finally, in cultured cells obtained from different areas of multinodular toxic goitres (N = 4), the response to TSH was similar to that of euthyroid goitres in cells prepared from 'cold' areas, and to that of toxic adenomas in cells obtained from 'hot' areas. The present data demonstrate the existence of an inhibitory action of unknown factors, possibly iodothyronines or thyroglobulin, on the TSH effect in short-term cultures obtained from thyrotoxic tissues. A normal TSH responsiveness can be restored when the culture is prolonged.

1988 ◽  
Vol 119 (3) ◽  
pp. 413-419 ◽  
Author(s):  
Mayumi Matsunaga ◽  
Katsumi Eguchi ◽  
Takaaki Fukuda ◽  
Hiroshi Tezuka ◽  
Yukitaka Ueki ◽  
...  

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.


1997 ◽  
Vol 82 (10) ◽  
pp. 3331-3336 ◽  
Author(s):  
Tsukasa Saito ◽  
Toyoshi Endo ◽  
Akio Kawaguchi ◽  
Masato Ikeda ◽  
Minoru Nakazato ◽  
...  

Abstract The Na+/I− symporter (NIS) is important in hormone synthesis in the thyroid gland. NIS activity, as reflected by I− uptake, was increased by TSH (1 mU/mL) or forskolin (10μ mol/L) in primary cultured human thyroid cells. Northern blot analysis revealed that incubation of these cells with TSH or forskolin for 24 h increased the abundance of NIS messenger ribonucleic acid (mRNA) 2.3- and 2.5-fold, respectively. Immunoblot analysis revealed 2.7- and 2.4-fold increases, respectively, in the amount of NIS protein after 48 h, suggesting that elevated levels of intracellular cAMP induced the expression of NIS in human thyrocytes. We then studied the levels of NIS mRNA and protein in Graves’ thyroid tissue and found that the amount of NIS mRNA in thyroid tissue from individuals with Graves’ disease (n = 5) was 3.8 times that in normal thyroid tissue (n = 5). The abundance of NIS mRNA was significantly correlated with that of thyroid peroxidase or thyroglobulin mRNAs, but not with that of TSH receptor mRNA, in the Graves’ and normal thyroid tissue specimens. The amount of NIS protein was also increased 3.1-fold in Graves’ thyroid tissue compared with that in normal thyroid tissue. The increased expression of NIS may thus contribute to the development of Graves’ disease.


1980 ◽  
Vol 86 (3) ◽  
pp. 443-449 ◽  
Author(s):  
B. ANDERBERG ◽  
S. ENESTRÖM ◽  
J. GILLQUIST ◽  
S. SMEDS

Analysis of the protein composition of the thyroid colloid was performed in 28 patients operated on for hyperthyroidism. Fifteen of the patients were treated before the operation with carbimazole combined with thyroxine and 13 were treated with propranolol alone. Colloid was collected by micropuncture of single follicles in peroperative thyroid biopsies. The protein composition was analysed by microgel electrophoresis and densitometry, both in the colloid samples and in the supernatant fraction of homogenates of microbiopsies from the thyroid specimens. The analyses showed that, during treatment with carbimazole and thyroxine, the relative amount of the larger thyroglobulin aggregates (S-TG) was decreased compared with the relative amount observed in the colloid from normal thyroid tissue. In the hyperfunctioning thyroid tissue from the propranolol-treated patients the protein composition of the colloid was similar to that observed in normal tissue and the relative amount of the S-TG fractions was significantly higher than in the carbimazole- and thyroxine-treated group. It may be concluded that the increased release of thyroid hormones in hyperthyroidism is not combined with changes in the protein composition of the thyroid colloid. The decreased relative amount of the S-TG fractions in the thyroid colloid from patients treated with carbimazole and thyroxine was probably due to an insufficient capacity to iodinate thyroglobulin.


1982 ◽  
Vol 95 (2) ◽  
pp. 237-244 ◽  
Author(s):  
C. A. Ollis ◽  
D. S. Munro ◽  
S. Tomlinson

We have described a system for the maintenance in culture of isolated human thyroid cells from both thyrotoxic tissue and non-toxic goitres. The cells isolated from the two thyroid tissue types showed similar cyclic AMP response characteristics to TSH with large increases in intracellular and extracellular cyclic AMP after 20-min incubations. Maximal responses were obtained with 50 mu. TSH/ml and half-maximal responses at 1·0 mu. TSH/ml. With cell passage the cyclic AMP responses to TSH decreased in magnitude and sensitivity. As with other thyroid cultures, growth of the cells with TSH induced arrangement into follicular structures, whereas cells grown in the absence of TSH remained as a monolayer. Basal intracellular cyclic AMP levels were increased in a dose-related fashion in cells grown in the presence of graded concentrations of TSH and the maximal response to further additions of TSH was not greater than in control cultures.


2001 ◽  
pp. 591-597 ◽  
Author(s):  
D Russo ◽  
S Bulotta ◽  
R Bruno ◽  
F Arturi ◽  
P Giannasio ◽  
...  

OBJECTIVE: The expression of two iodide transporters, the sodium/iodide symporter (NIS) and pendrin, was analyzed in thyroid tissues of patients with toxic multinodular goiter (TMNG) and non-toxic multinodular goiter (MNG). METHODS: The levels of NIS and pendrin proteins were analyzed in total protein extracts from nodular and non-nodular tissues by Western blot. RESULTS: In tissue samples from TMNG, we found an increased expression of NIS (2.5-fold) in the hot nodules, and similar levels between cold nodules and non-nodular tissues. In contrast, the levels of pendrin were slightly increased in both hot and cold nodules from TMNG, and decreased (about twofold) in cold nodules from MNG. We also noticed that there was no relationship between NIS and pendrin expression. CONCLUSIONS: Our data demonstrate that hot nodules from TMNG express a higher number of iodide transporters (mainly NIS), whereas cold nodules from TMNG, but not from MNG, show levels of the two proteins comparable with normal tissue, suggesting a role in vivo of TSH in maintaining the expression of NIS and pendrin protein in normal thyroid tissue. Finally, different mechanisms are involved in the regulation of NIS and pendrin expression.


1981 ◽  
Vol 98 (3) ◽  
pp. 370-376 ◽  
Author(s):  
Stephen P. Bidey ◽  
Nicholas J. Marshall ◽  
Roger P. Ekins

Abstract. The cyclic AMP response to thyrotrophin (TSH) has been investigated in cells prepared from human thyroid tissue obtained during surgery for sub-total laryngectomy, and maintained under in vitro conditions as primary monolayer cultures. When cells were incubated with 1.0 mU TSH/ml, a maximal level of intracellular cyclic AMP was reached after 20 min of incubation in the presence of 0.5 mm 3-isobutyl-1-methyl xanthine (MIX). This level of cyclic AMP was sustained for at least 2 h. Half-maximal stimulation of cyclic AMP was produced by TSH doses of between 1 and 5 mU/ml. In a study of a series of eight groups of monolayer cultures, each derived from a single, different thyroid gland, the mean stimulation of cyclic AMP given by 50 mU TSH/ml was 37.8-fold greater than in non-stimulated cell monolayers. Significant stimulation to 50 μU TSH/ml was observed in some monolayers and the precision of measurement of TSH was better than 15% over the TSH dose range 0.2–1.0 mU/ml. The magnitude of the cyclic AMP response to TSH was unaffected by the presence in the incubation medium of 20% (v/v) normal human serum. A cyclic AMP response to TSH was still demonstrable in cells that had been maintained for a period of 22 days in monolayer culture, although the response was reduced in comparison with that given by 4–5 day old cultures.


1987 ◽  
Vol 113 (3) ◽  
pp. 403-412 ◽  
Author(s):  
S. Y. Chow ◽  
Y. C. Yen-Chow ◽  
H. S. White ◽  
D. M. Woodbury

ABSTRACT Iodide uptake by primary cultures of turtle thyroid follicular cells is directly proportional to the Na + concentration and is inversely proportional to the HCO3− concentration in culture medium, but is not affected by the Cl− concentration. Addition of 4,4′-di-isothiocyano-2,2′-stilbene disulphonate (DIDS; 10 μmol/l and higher doses) to medium containing different concentrations of Na+ (5–140 mmol/l), HCO3− (0–40 mmol/l) and Cl − (120 mmol/l) generally enhanced iodide uptake by the cultured cells; however, there was no significant effect in Na+-free and in low Cl− (90 mmol/l and less) medium. The inhibitory effects on iodide uptake of ouabain, frusemide and perchlorate were attenuated by DIDS which also antagonized the stimulatory effects on iodide uptake of TSH, although both DIDS and TSH increased the 125I− cell/medium ratio when they were given alone. At doses of 100 μmol/l and higher, DIDS lowered the intracellular pH of cultured cells when the pH of the medium was maintained at a constant level. It also increased the intracellular Cl − concentration, but had no effect on intracellular Na+ or K +. The input and specific resistances of cell membranes in cultured thyroid cells and in isolated thyroid slices increased (decreased conductance) after adding DIDS to the perfusion fluids. Both Na+/K+- and HCO3−-ATPase activities in homogenates of turtle thyroid tissue were inhibited by DIDS. Results from this investigation demonstrate (1) that in addition to preventing the leak of iodide from thyroid cells, DIDS may act to increase the sensitivity of the Na + -anion carrier to I− and thereby increases iodide uptake, and (2) that a HCO3−–Cl− exchange system is present in the thyroid cell membrane and appears to be linked to the transport of iodide into thyroid cells. J. Endocr. (1987) 113, 403–412


1964 ◽  
Vol 62 (4) ◽  
pp. 413-424 ◽  
Author(s):  
Anne E. Caunt ◽  
D. Taylor-Robinson

Infected human thyroid tissue cultures have been found to yield free V-Z virus on ultrasonic disintegration and some virus is also present in the fluid phase of intact tissue cultures, while the yield of virus from infected human amnion cells is very low and no free virus is found in the fluid phase. Thyroid cultures are more susceptible to infection with V-Z virus, and the infected cultures are a better source of complement-fixing antigen, than are amnion cultures.The lack of free virus in infected amnion tissue cultures cannot be attributed to protection by interferon. It also seems likely that if any infectious virus were released it would be more likely to be adsorbed to normal cells than to be inactivated, but as no secondary foci appear in amnion monolayers we conclude that infective virus is not released into the medium.It is suggested that the virus in thyroid cells is present in a more complete form than that in amnion cells.We are grateful to Dr R. M. Rawcliffe of Birkenhead General Hospital and Dr F. Whitwell of Broadgreen Hospital, Liverpool, for arranging the supply of thyroid tissue.


Author(s):  
R. D. Sjolund ◽  
C. Y. Shih

The differentiation of phloem in plant tissue cultures offers a unique opportunity to study the development and structure of sieve elements in a manner that avoids the injury responses associated with the processing of similar elements in intact plants. Short segments of sieve elements formed in tissue cultures can be fixed intact while the longer strands occuring in whole plants must be cut into shorter lengths before processing. While iyuch controversy surrounds the question of phloem function in tissue cultures , sieve elements formed in these cultured cells are structurally similar to those of Intact plants. We are particullarly Interested In the structure of the plasma membrane and the peripheral ER in these cells because of their possible role in the energy-dependent active transport of sucrose into the sieve elements.


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