Thyrotrophin Displacement Activity of Serum Immunoglobulins in Health and Disease

Summary: Thyrotrophin displacement activity of serum immunoglobulins in health and disease. M. S. Croxson, T. M. T. Lim, F. M. Graham and H. K. Ibbertson, Aust. N.Z. J. Med ., 1980, 10, pp. 151–156.
A radioreceptor assay for TSH displacement activity (TDA) was employed as an index of thyroid stimulating immunoglobulins (TSIg) in serum from patients with autoimmune thyroid disease. TDA was > 20 in normal subjects (37/37), patients without thyroid disease (36/38), non-toxic goitre (18/19) and toxic nodular goitre (16/17) and was significantly lower in Nepalese subjects with severe endemic goitre. TDA was > 20 in 84% of patients with untreated hyperthyroid Graves' disease (70/83), in Hashimoto's thyroiditis (3/17) and following subtotal thyroidectomy (8/14) or ¹³¹I therapy (7/11), but was ≤ 20 in euthyroid patients treated with carbimazole (58/59) and during spontaneous remission (18/19). Serial TDA measurements during antithyroid drug therapy suggested that carbimazole has a direct effect in reducing TS/g concentration but that TDA is not a useful prognostic indicator of short term relapse or remission     

COMPARATIVE EVALUATION OF CYCLIC AMP AND IODIDE ACCUMULATION RESPONSES TO THYROID-STIMULATING IMMUNOGLOBULINS IN CULTURED FRTL-5 CELLS

The rat thyroid cell strain FRTL-5 was used to investigate the relationship between cyclic AMP and iodide accumulation responses to thyroid-stimulating immunoglobulins (TSIg). Immunoglobulin G-enriched precipitates of sera from 19 of 21 (90%) newly-diagnosed Graves' disease patients gave significant (P less than 0.01) accumulation of iodide (125I), and 16 of these also stimulated intracellular cyclic AMP. Correlation was poor however, with certain TSIg preparations giving widely divergent responses. After initiation of antithyroid treatment, 40% of sera investigated contained TSIg detectable in both bioassay systems, and all but one of the remainder were stimulatory in one of the two bioassays. All patients in remission were devoid of detectable TSIg as determined by iodide accumulation, although a single preparation stimulated cyclic AMP accumulation. LATS-B, a lyophilized reference serum preparation containing high TSIg activity, enhanced iodide accumulation, which showed evidence of correlation with intracellular cyclic AMP at doses above 0.5 mU/ml. At lower doses, iodide accumulation was observed in the absence of detectable cyclic AMP accumulation. TSH and LATS-B-induced iodide accumulation were enhanced, and iodide efflux reduced, by the anion channel blocker 4-4' diisothiocyanate stilbene 2,2' disulphonic acid (DIDS). In contrast, Ig-enriched fractions of normal sera decreased both basal and stimulated iodide accumulation, but were without effect on efflux. TSIg from untreated Graves' sera gave widely-differing iodide accumulation responses which showed poor correlation with both intracellular cyclic AMP and cyclic-AMP-independent iodide efflux. This clear dissociation of responses to serum Ig preparations suggests that iodide uptake in FRTL-5 cells, which do not organify iodide, may be subject to variable effects of non-TSIg components of Graves' sera, on both iodide uptake itself, and as inhibitors of TSIg-induced accumulation of intracellular cyclic AMP.     

The effects of dopaminergic blockade on serum TSH and prolactin levels in thyrotoxicosis

In the first part of this study, we have demonstrated that, in 7 patients with untreated thyrotoxicosis, a 7 day regime of the long acting dopamine antagonist metoclopramide (10 mg orally 8 hourly) produces more adequate dopaminergic blockade at pituitary level than a single oral 10 mg dose of the compound as assessed by serum prolactin responses. Subsequently, we have employed this protracted oral metoclopramide regime to evaluate the contribution of dopaminergic tone to the abnormal TSH and prolactin responsiveness of thyrotoxicosis. Serum TSH and prolactin responses to iv TRH (200 micrograms) were measured in 10 untreated thyrotoxic patients before and after a 7 day period of metoclopramide 10 mg orally 8 hourly. Ten euthyroid individuals were studied in similar fashion, their serum samples being analysed for prolactin levels alone, thus providing a control group for prolactin responsiveness to TRH, before and after metoclopramide. In the thyrotoxic patients basal TSH levels did not change as a consequence of metoclopramide therapy and the TSH response to TRH remained flat. Basal prolactin levels were similar in thyrotoxic and euthyroid individuals and the increase in prolactin, seen in both groups after metoclopramide, was smaller in the thyrotoxic group than in the euthyroid group. Prolactin responsiveness to TRH was significantly impaired in the thyrotoxic subjects as compared to euthyroid subjects. After metoclopramide there was a significant decline in prolactin responsiveness in the euthyroid group, and a similar, though insignificant, trend in the thyrotoxic patients. We conclude that in thyrotoxicosis dopaminergic tone plays no major part in the suppression of TSH levels, nor in the impaired prolactin responsiveness to TRH.     

Effects of human thyroid-stimulating hormone and immunoglobulins on adenylate cyclase activity and the accumulation of cyclic AMP in human thyroid membranes and slices.

The activation of adenylate cyclase and the accumulation of cyclic AMP resulting from the action of human thyroid-stimulating hormone (TSH), long-acting thyroid stimulator (LATS) or LATS-protector (LATS-P) have been investigated in preparations of human thyroid membranes and slices. Human TSH significantly increased adenylate cyclase activity in membranes from non-toxic goitres whereas LATS and LATS-P had no consistent effect. However, pre-incubation of goitrous membranes with LATS--immunoglobulin G inhibited the effect of TSH on adenylate cyclase. When thyroid membranes were prepared from the glands of patients with Graves's disease neither TSH nor thyroid-stimulating immunoglobulins (TSIg) stimulated adenylate cyclase significantly. Whether from non-toxic goitres or thyrotoxic tissue, the concentration of TSH needed to induce half of the maximum response was lower in thyroid slices than in membranes. Both LATS and LATS-P significantly stimulated the accumulation of cyclic AMP in slices of goitrous tissue but thyrotoxic tissue slices did not respond. In goitrous slices, submaximum concentrations of TSH and TSIg caused additive responses in the accumulation of cyclic AMP but TSIg did not increase the maximum response to TSH.     

Comparison of human and porcine thyroid membranes for radioreceptor assay of bovine thyrotrophin and thyrotrophin-binding inhibiting immunoglobulins

A detailed comparison between the use of human and porcine thyroid membranes for the radioreceptor assay (RRA) of bovine TSH (bTSH) and thyrotrophin-binding inhibiting immunoglobulins (TBIIg) is reported. Bovine thyroid membranes were also investigated but were found to be far less satisfactory than either human or porcine thyroid membranes. The affinity constant (ka) of the interaction of bTSH with porcine thyroid membranes (Ka = 3.3 X 10(9) l/mol) measured b Scatchard analysis was higher than with human thyroid membranes (Ka = 2.1 X 10(8) l/mol). Porcine thyroid membranes were more sensitive for the assay of bTSH (detection limit 30 microunits, half-maximal inhibition 0.3 microunit) than human thyroid membranes (detection limit 200 microunits, half-maximal inhibition 7.4 mu.). Preincubation of membranes from either species with immunoglobulin rich in long-acting thyroid stimulator (LATS) inhibited the saturable binding of 125I-labelled TSH to a greater extent than did normal immunoglobulin. The binding of 125I-labelled TSH to porcine membranes was more sensitive to the inhibitory effect of LATS-immunoglobulin and was also less affected by normal immunoglobulin than was binding to human thyroid membranes. When assayed with each type of membrane preparation there was good correlation between the RRA of immunoglobulins prepared from patients with Grave's disease and from normal subjects (n = 18) (r = 0.85, P less than 0.001, n = 73). The incidence of positive TBIIg in untreated Grave's disease was greater for porcine than for human thyroid membranes.     

Increased plasma 5 alpha-androstane-3 alpha,17 beta-diol glucuronide concentration in clinically euthyroid women with suppressed plasma thyrotropin levels: further evidence for generalized tissue overexposure to thyroid hormones in these subjects.

It has been reported that plasma levels of 5 alpha-androstane-3 alpha,17 beta-diol glucuronide (ADG) are increased in thyrotoxic patients. The present study was carried out to determine whether in clinically euthyroid subjects with suppressed TSH levels, ADG plasma levels would also be increased, suggesting a more generalized tissue overexposure to thyroid hormones. The latter has been suggested from increased sex hormone-binding globulin levels (SHBG) in these subjects. ADG and SHBG levels were measured in a group of 20 euthyroid postmenopausal women, 20 postmenopausal women with clinically evident thyrotoxicosis, and 16 euthyroid postmenopausal women with suppressed TSH levels. The mean ADG level in the thyrotoxic group was 4 times the mean level in the control group, whereas in the group with isolated TSH suppression the mean level was about twice the level in the control group. As levels of dehydroepiandrosterone sulfate, the major precursor of plasma ADG, were not significantly increased in these subjects, these data suggest an increased conversion of dehydroepiandrosterone sulfate to ADG. Together with the increased SHBG levels observed in both thyrotoxic and euthyroid subjects with suppressed serum TSH concentrations, these data support the concept that the TSH-suppressed euthyroid subjects have generalized increased thyroid hormone effects on peripheral tissues.     

BIOASSAY OF THYROID-STIMULATING IMMUNOGLOBULINS USING HUMAN THYROID CELL CULTURES: OPTIMIZATION AND CLINICAL ASSESSMENT

Primary monolayer cultures of human thyroid cells have been used to investigate the intracellular cyclic AMP response to thyroid-stimulating immunoglobulins (TSIg) prepared from the sera of patients with Graves' disease. In particular, attention was directed towards dose-response characteristics obtained under differing incubation conditions, and an optimized incubation procedure based upon conditions consistent with the maximal precision of TSIg measurement was developed. The magnitude of the cyclic AMP response to a tested TSIg preparation was shown to be greatest after the longest incubation period investigated, 16 h, for all doses of TSIg investigated. The greatest precision of TSIg measurement, as determined by the lowest relative error, was obtained at an immunoglobulin (Ig) dose of 1 mg/ml, and when incubated at this dose level for 16 h, all Ig-enriched fractions prepared from the sera of 28 newly-diagnosed patients with Graves' disease significantly (P less than 0.05) raised the intracellular cyclic AMP level of thyroid cell monolayers. After thionamide drug therapy, the incidence of TSIg detection declined to 42% (5 of 12 patients). TSIg bioactivity was also found in 9 of 14 (64%) patients with euthyroid exophthalmos and in 4 of 11 (36%) cases of non-toxic goitre. In contrast, Ig's prepared from the sera of normal euthyroid volunteers were devoid of TSIg bioactivity.     

Changes in circulating thyroid hormone levels and systolic time intervals in acute hypothyroidism

OBJECTIVE: We have previously reported that, in thyrotoxic patients treated with carbimazole, serum T4 and T3 levels are the first parameters to return to normal, followed by the systolic time interval (STI, a marker of thyroid function at tissue level) and then the serum TSH. The aim of this study was to compare the rate of change of thyroid hormones, TSH and STI in treated hypothyroid patients after the sudden withdrawal of thyroxine. DESIGN AND PATIENTS: Serum T4, T3 (free and total) and TSH were measured in 12 patients taking thyroxine for primary hypothyroidism; seven were biochemically euthyroid and five were over-replaced, as defined by an elevated free T4 and a sub-normal TSH. Thyroxine was withdrawn and the measurements repeated three times a week until the STI rose above the euthyroid range (0.26-0.32). RESULTS: After stopping thyroxine, the serum TSH and STI left the normal range, in advance of the free T4 and T3, after 9.5 +/- 0.95 and 12.2 +/- 1.5 days respectively (mean +/- SEM). The TSH was the first parameter to leave the euthyroid range in all subjects except one in whom the serum TSH was fully suppressed (less than 0.05 mU/l) initially. In the euthyroid group the TSH and STI increased rapidly after stopping thyroxine (time to leave euthyroid range 7.4 +/- 0.8 and 9.4 +/- 0.7 days respectively). In contrast, in the over-replaced group serum TSH and STI became elevated after 12.4 +/- 1.0 days (P less than 0.005 vs euthyroid group) and 16.0 +/- 2.7 days (P less than 0.05 vs euthyroid group) respectively. There was no delay in the fall in serum T4 or T3 in the over-replaced group when compared with the euthyroid group. CONCLUSIONS: In the evolution of primary hypothyroidism, markers of thyroid function at a tissue level (TSH and STI) become abnormal in advance of thyroid hormones. After stopping thyroxine therapy in treated hypothyroid patients, there is a delayed rise in STI and serum TSH levels in subjects with a subnormal TSH level, as compared with those with a normal TSH on treatment. This suggests mild tissue thyrotoxicosis in these individuals.     

EFFECTS OF LUGOL''S SOLUTION ON THYROID FUNCTION IN NORMALS AND PATIENTS WITH UNTREATED THYROTOXICOSIS

Thirty-eight normal volunteers and 10 patients with untreated thyrotoxicosis were each given 0.5 ml of Lugol's solution daily for 10 days. On days 0, 5, 10, 15 and 20, serum levels of T4, free T4, T3 and TSH (by sensitive immunoradiometric assay) were measured. In normal subjects, the serum concentrations of free T4 declined significantly at day 10 while TSH levels were significantly increased at days 5, 10 and 15. Serum levels of T4 and T3 did not change significantly. All the observed changes took place within the limits of normal ranges for the hormones mentioned. In contrast, in the thyrotoxic subjects, both T4 and T3 were significantly decreased at days 5 and 10, while serum TSH remained below detection limit (0.14 mU/l) throughout the study. Short exposure to excessive iodide in normal subjects affects T4 and T3 release and this effect could be partially overcome by compensatory increase in TSH. In thyrotoxicosis, lack of compensatory increase in TSH results in rapid decreases in T4 and T3 levels. The integrity of the hypothalamo-pituitary-thyroidal axis may be effectively assessed by measuring TSH response to iodide suppression, using a highly sensitive immunoradiometric assay.     

Thyrotropin displacement activity of serum immunoglobulins from patients with Graves' disease.

A radioreceptor assay for human thyroid stimulators has been employed in various groups of patients. The ability of Ig to displace the labeled TSH from the receptors is referred to as "TSH displacement activity (TDA)." In active Graves' disease, TDA was above normal in 76% of the cases, and in the remaining patients, it was above the 76th percentile, suggesting that thyroid-stimulating Ig (TSIg) may have been present in all cases, but not always demonstrable by this method. Significant TDA was not found in normal persons or in toxic or nontoxic nodular goiters. It was also negative in some patients with "euthyroid ophthalmic Graves' disease." In patients with Graves' disease controlled with antithyroid drugs, positive TDA accurately predicted the recurrence of hyperthyroidism in eight of nine cases from whom the drugs were withdrawn. Thus, TSIg appears to be a prerequisite of the hyperthyroidism of Graves' disease. Moreover, the remission of hyperthyroidism was due to the disappearance of TSIg (immunological remission) in most cases in this small series. Serum TDA may provide a means of detecting immunological remission. The exophthalmos of Graves' disease does not require thyroid-stimulating Ig.     

Studies on the radioreceptor assay of TSH: the properties of TSH-binding inhibitor immunoglobulins (TBII) in patients with Graves' disease (author's transl)]

In the radioreceptor assay system for TSH, serum immunoglobulin G (IgG) from some patients with Graves' disease has been shown to inhibit the binding of labelled TSH to its receptor sites. In order to clarify the properties of these TSH-binding inhibitor immunoglobulins (TBII) in patients with Graves' disease, TBII were measured in sera from 31 untreated and 51 131I-treated patients, and their relation to clinical and laboratory findings was studied. TBII were detected in 18 (60%) out of 31 patients with untreated Graves' disease. TBII levels in these patients correlated well with thyroidal 99mTc uptake at 30 min and also with the grade of epithelial hyperplasia of thyroid follicles. There was no significant correlation between TBII and serum T3, serum T4, free T4 index, antibody titers against thyroglobulin and microsomes, or association of exophthalmos. There were many patients with Graves' disease whose sera contained high TBII levels but no detectable bioassayable thyroid-stimulating activity (LATS), and in these patients a close correlation was observed between serum levels of TBII and bioassayable LATS-protector activity. In patients with Graves' disease who had been treated by 131I from 5 to 17 years before, the incidence of TBII was very low at 20% (10/51). All except two cases having TBII were found to be still thyrotoxic. Thus, TBII were detected in 8 out of 10 thyrotoxic patients and in only 2 out of 18 euthyroid and none of 23 hypothyroid patients. These findings suggest that TBII in patients with Graves' disease were in close association with human thyroid stimulating activity, and that TBII might be useful as an indicator for checking the effectiveness of the treatment.     

Serum protein inhibition of thyrotropin binding to human thyroid tissue.

We used a modification of the TSH radioreceptor assay to detect TSH-binding inhibition (TBI) activity in serum and serum fractions from normal subjects and patients with Graves' disease. TBI activity is present in normal IgG prepared by DEAE-Sephadex chromatography and in normal globulins prepared by precipitation at 1.6 M ammonium sulfate. Other normal serum proteins also had TBI activity when large concentrations were tested. Gel filtration chromatography and powder block electrophoresis were used to prepare fractions of normal and Graves' disease sera. In these fractions from normal serum. TBI activity was found in both gamma-globulin and alpha-globulin-albumin fractions electrophoretically and in both 7S and 4S peaks from gel filtration. TBI activity from Graves' disease patients' sera was similarly distributed, but relatively more TBI accompanied the electrophoretic gamma-globulins. Sepharose Protein-A and anti-IgG were used as immunoabsorbents to isolate and purify IgG from normal and Graves' disease sera. TBI activity in IgG was proportional to the IgG concentration, indicating that the TBI which migrates as a gamma-globulin electrophoretically is an IgG and thus may possibly be an antibody. Inhibitory activity found in normal serum globulins and the non-IgG fractions of both normal and abnormal sera seriously interferes with attempts to use the TSH radioreceptor assay to study the hypothesized anti-TSH, receptor antibody in the serum of patients with Graves' disease.     

Euthyroid Graves' disease showing no thyroid abnormalities except positive thyroid-stimulating antibody (TSAb): two case reports

Abstract. We report two cases of euthyroid Graves' disease in women who had ophthalmopathy without previous history of hyperthyroidism. Enlargement of extraocular muscles was observed by magnetic resonance imaging (MRI). The patients had no thyroid enlargement and their serum concentrations of free T₄, free T₃, and TSH were normal. The sera were negative for antithyroid microsomal and thyrogobulin antibodies, and anti-TSH receptor antibodies measured by radioreceptor assay. T₃ suppression test results were normal. Only thyroid-stimulating antibody (TSAb) measured by sensitive bioassay was positive. These findings indicate that sensitive TSAb is the most useful laboratory test in the diagnosis of euthyroid Graves' disease.     

Interaction of the mouse thyrotrophin receptor with thyrotrophin binding inhibitor immunoglobulins

The species-specificity of thyrotrophin binding inhibitor immunoglobulin (TBII) for the thyroid TSH receptor was investigated using a preparation of thyroid plasma membranes (TPM) from propylthiouracil-treated mice, as well as from human glands. The interest in the mouse arose from its use as the bioassay animal for the long-acting thyroid stimulator (LATS). A comparison was made of the response in the two radioreceptor assays of serum immunoglobulins from ten normal subjects and twenty patients with Graves's disease, who had also been selected to have positive TBII activity in the assay based on human TPM. All the specimens from the patients with Graves's disease had detectable TBII activity in the mouse radioreceptor assay, inhibiting the binding of 125I-labelled TSH to a greater extent than did any of the specimens from normal subjects. There was evidence for a minor degree of species-specificity, since at least one of the specimens from the Graves' disease group had unexpectedly high activity in the assay based on mouse TPM and another had unexpectedly weak activity in that assay. However, this specificity appeared to be unrelated to the presence or absence of LATS. The effect of LATS on the response of serum immunoglobulins in the mouse radioreceptor assay was tested using nine patients with Graves's disease who had undetectable serum LATS and another eight patients with Graves's disease whose serum gave a positive LATS response. These patients had also all been selected to have positive TBII activity in their serum, as determined with human TPM. All samples from each of the LATS-positive and LATS-negative subgroups gave a positive TBII response in the radioreceptor assay based on mouse TPM, and there was extensive overlap between the individual values for the two subgroups. It is concluded that the failure of some TBII-positive serum immunoglobulins to stimulate the mouse thyroid gland and produce a positive LATS response is not due to species-specificity at the level of receptor binding.     

Thyrotrophin and thyroid-stimulating immunoglobulins have similar characteristics in activating human thyroid membrane adenylate cyclase

The activation of adenylate cyclase by TSH and thyroid-stimulating immunoglobulins (TSIg) was investigated in membranes prepared from non-toxic goitres. The assay conditions for maximal adenylate cyclase stimulation by TSH in the absence and presence of a cell cytosol preparation were determined. Cell cytosol had no detectable effect on the characteristics of activation by TSH although it increased the production of cyclic AMP in response to the hormone. In the presence of cell cytosol, membrane protein, pH and substrate concentration dependency of adenylate cyclase activation by both TSH and TSIg were similar. In a comparison of the time-courses of activation there was no apparent lag phase in the activation of adenylate cyclase by TSIg but the onset of activation was slower and linearity persisted longer than with TSH.     

Hyperthyroid Graves' disease without detectable thyrotropin receptor antibodies.

TSH receptor antibodies are generally held responsible for the stimulation of the thyroid that characterizes patients with Graves' disease. Here, we describe nine patients with hyperthyroidism (triiodothyronine 4.9, 3.2-6.7 nmol/L; median, range) who were referred for radioiodine treatment and who had increased thyroid radioiodine uptake values but lacked TSH receptor antibodies determined by a radioreceptor assay. Furthermore, when serum immunoglobulins were studied in a bioassay based on a rat thyroid cell line (FRTL-5), no evidence of stimulant activity was observed. Subsequent to radioiodine therapy, TSH receptor antibodies appeared in all nine patients. The antibodies competed for TSH in the radioreceptor assay and, of the eight patient samples studied with the bioassay, six stimulated cAMP production whereas another two blocked the latter. The results show that a small proportion of patients with active hyperthyroid Graves' disease, in this study 9 out of 130 cases, do not have detectable TSH receptor stimulatory antibodies. A local production of antibodies within the thyroid can be suggested, although a more likely explanation might be that the thyroid in Graves' disease is activated also by other mechanisms than antibody-dependent ones.     

HIGHER THAN CONVENTIONAL DOSES OF CARBIMAZOLE IN THE TREATMENT OF THYROTOXICOSIS

In order to ascertain whether higher than conventional doses of carbimazole achieve more rapid control of thyrotoxicosis, 30 thyrotoxic patients were alternately allocated into two groups, group 1 (15 subjects) receiving a conventional starting dose of 45 mg orally daily and group 2 (15 subjects) a dose of 100 mg orally. In addition to weekly estimations of serum T4, T3, free T4, free T3 and TSH, the systolic time intervals ratio (STI), a measure of left ventricular contractility, was calculated as an accurate measure of peripheral thyroid hormone activity, the study end-point being a normal STI (0.26-0.32). None of the individuals studied experienced side-effects during the study period. Mean pre-treatment STI values for the two treatment groups were the same at entry (0.20). The mean recovery times for STI was 4.4 weeks (SE 0.3) in the high dose group and 5.9 weeks (SE 0.4) in the low dose group (P = 0.0037). There was a definite trend towards a shorter recovery time for free T3 in the higher dose group (P = 0.057) but no apparent differences for T4, T3 and free T4. Higher than conventional doses of carbimazole may be advisable in the initial treatment of severe thyrotoxicosis.     

Thyroid stimulating immunoglobulin bioactivity during carbimazole therapy as measured by the cytochemical bioassay

Most assays of thyroid stimulating immunoglobulin (TSI) are unsuitable for the quantitation of TSI during the treatment of Graves' hyperthyroidism because assay insensitivity results in some negative responses. Therefore the sensitive cytochemical bioassay was used to investigate the effect of carbimazole on TSI levels as a possible mechanism for the induction of the increased remission rate which is characteristic of thionamide therapy. Twelve patients were studied before therapy for de-novo Graves' hyperthyroidism; seven patients consented to a detailed prospectively study during block-replace therapy with carbimazole 10mg 6 hourly with a later addition of T3 20 micrograms 6 hourly when biochemically euthyroid. In addition thyroid hormone or T3 suppressed technetium (99m Tc) thyroidal uptake was monitored at between weekly and 3 monthly intervals, as well as the clinical findings, total T4 total T3, TSH, antimicrosomal antibody titers and immunoglobulins IgG, IgM and IgA. TSI was detected in all patients before treatment but there was no correlation with any other pretreatment measurements. During therapy TSI fell (in three different patterns) in 6 out of 7 patients studied for between 14-55 weeks (mean 29 weeks). TSI remained unchanged in one patient. Only the 99m Tc uptake correlated with TSI activity in the treated patients as a group (r = 0.71, p less than 0.001). TSI remained detectable in all patients, even in 4 patients in whom T3 suppression of 99m Tc was demonstrated. There is some evidence for a carbimazole effect lowering TSI activity, however relapse rate did not support this. T3 suppressed 99m Tc uptake may be a sensitive in vivo marker of TSI activity.     

Retinol has specific effects on binding of thyrotrophin to cultured porcine thyrocytes

Retinoids are potential candidates for the treatment of thyroid cancer. However, one of the disadvantages of these substances is their dedifferentiating effect on normal non-transformed thyrocytes. To identify conditions under which no dedifferentiating effect of retinol on normal thyrocytes can be observed, we determined iodide uptake, protein iodination, expression of sodium-iodide symporter (NIS) mRNA and protein, and the binding of iodine-125-labelled bTSH in cultured porcine thyrocytes. Combination of TSH and < or =6.5 microM retinol increased iodide uptake and protein iodination compared with TSH alone over the entire incubation time, whereas TSH plus > or =13 microM retinol increased the uptake of iodine-125 only during the first 12 h but decreased it after 30 h and longer. After > or =30 h incubation times with > or =13 microM retinol, the fraction of apoptotic cells was enhanced and proliferation decreased. The incubation with retinol enhanced the binding of [125I]bTSH to thyrocytes, but did not influence expression of the NIS. With low retinol concentrations, the effect on the binding of TSH apparently predominated and retinol increased thyroid function; with higher concentrations the pro-apoptotic effect of retinol overlapped and a two-phased time course resulted. It can be concluded that low concentrations of retinol also exert differentiating effects in normal thyrocytes.     

Studies on the radioreceptor assay of TSH: the properties of TSH-binding inhibitor immunoglobulins (TBII) in patients with Hashimoto's thyroiditis (author's transl)]

TSH-binding inhibitor immunoglobulins (TBII) have been detected not only in patients with Graves' disease but also in those with Hashimoto's thyroiditis by using the radioreceptor assay of TSH. In the present study, the properties of TBII in patients with Hashimoto's thyroiditis are discussed. Two (7%) of 29 patients with Hashimoto's thyroiditis had detectable levels of TBII in their gamma-globulin fractions. Both patients were untreated and clinically hypothyroid. One of them had no goiter, and her thyroidal 99mTc uptake was 0% (normal range: 0.4 approximately 3.0%). Despite having potent TSH-binding inhibitor activity in the TSH radioreceptor assay, her serum or its IgG fraction (H-IgG) did not contain any significant anti-TSH antibody, LATS, LATS-protector or human thyroid adenylate cyclase (AC) stimulating activity. This H-IgG inhibited both human thyroid AC stimulation and c-AMP increase in mice thyroid glands induced by TSH or LATS. Furthermore, her serum caused significant inhibition of 131I-release by LATS in a McKenzie mouse bioassay. The present study demonstrates that the serum of one patient with Hashimoto's thyroiditis contained antibodies which 1) blocked the binding of lablled TSH to the receptor, 2) had no thyroid-stimulating activity by themselves, and 3) inhibited AC stimulation by TSH. Such antibodies may cause unresponsiveness to TSH stimulation, hypothyroidism, and, if this state persists for a long time, eventually may result in atrophy of the thyroid tissue. Further, these data indicate that TBII detected by the TSH radioreceptor assay did not always show thyroid stimulating activities.