Highly specific antisera to triiodothyronine (T(3)) were prepared by immunization of rabbits with T(3)-bovine serum albumin conjugates. Antisera with T(3) binding capacity of up to 600 ng/ml were obtained. The ability of various thyronine derivatives to inhibit the binding of T(3-) (125)I to anti-T(3) serum was found to vary considerably. l-T(3), d-T(3) and several triiodoanalogues were potent inhibitors of the reaction. Little inhibition of T(3-) (125)I binding was produced by l-thyroxine (T(4)) or other tetraiodo- analogues, thyronine or iodotyrosines. Chromatography of several T(4) preparations indicated that most of their very slight activity could be ascribed to contamination with T(3). Successful assay of T(3) in serum was accomplished by the addition of diphenylhydantoin to the assay system. Under these circumstances, recovery of T(3) added to serum was excellent, and addition of T(4) was without significant effect. Serum T(3) concentrations in normal subjects averaged 145 +/-25 ng/100 ml (sd). Increased concentrations (429 +/-146 ng/100 ml) were observed in hyperthyroid patients whereas those with hypothyroidism had serum T(3) levels of 99 +/-24 ng/100 ml. Elevated T(3) concentrations were found also in hypothyroid patients receiving 25 mug or more of T(3) daily and in those receiving 300 mug of T(4) daily. Serial measurements of T(3) concentrations in subjects after oral T(3) administration revealed peak T(3) concentrations 2-4 hr after T(3) administration. Intramuscular administration of thyrotropin (TSH) resulted in earlier and more pronounced increases in serum T(3) than in serum T(4) concentrations. Triiodothyronine (T(3))(1) was recognized to be a biologically active secretory product of the thyroid gland over a decade ago (1). Recent studies have indicated that it is formed extrathyroidally as well (2, 3). Nevertheless, relatively little information concerning the role of T(3) secretion in different thyroid disorders has been accumulated until very recently. Methods for the measurement of T(3) which require its extraction from plasma, and often its separation from thyroxine as well, have been described by several investigators (4-11). These methods have proven useful, but they are relatively complicated, the number of samples that can be assayed is limited, and they may be affected by in vitro deiodination of thyroxine. More recently the radioimmunoassay technique has been applied to the measurement of T(3). Several preliminary reports have appeared describing the preparation of antibody to triiodothyronine by immunization of animals with T(3)-protein conjugates and its use for the measurement of T(3) in serum (12-15). The present report describes the development of a radioimmunoassay for the measurement of T(3), studies of the specificity of the anti-T(3) serum, and some initial studies which indicate that the method is applicable to the measurement of T(3) in unextracted serum.

Animals↗

PubMed1972

Inhibition of thyrotropin response to thyrotropin-releasing hormone by small quantities of thyroid hormones.

Inhibition of thyrotropin (TSH) release by chronic treatment with small quantities of triiodothyronine (T(3)) and thyroxine (T(4)) was evaluated by determining the serum TSH response to thyrotropin-releasing hormone (TRH) in normal subjects and hypothyroid patients. Response to TRH was determined before treatment and after each dosage of a synthetic combination of T(3) + T(4) had been given for 3-4 wk. Treatment of eight normal subjects with 15 mug T(3) + 60 mug T(4) reduced the maximum increase in serum TSH above baseline (maximum DeltaTSH) by 76% in response to 400 mug TRH and by 87% in response to 25 mug TRH. The average serum T(3) level during a 24 hr period in normal subjects who had been taking 15 mug T(3) + 60 mug T(4) for 3-4 wk was 129+/-10 ng/100 ml (mean +/-SEM), well within the normal range, 70-150 ng/100 ml, although higher than the pretreatment level, 98+/-7 ng/100 ml. The average serum T(4) level was unchanged from the pretreatment level. Treatment of the same subjects with 30 mug T(3) + 120 mug T(4) reduced the maximum DeltaTSH further.Six patients with primary hypothyroidism were treated, sequentially, with 15 + 60, 22.5 + 90, and 30 mug T(3) + 120 mug T(4). For each patient there was one increase in dosage of 7.5 mug T(3) + 30 mug T(4) which abruptly converted a maximum DeltaTSH that was greater than, or at the upper limit of, normal to one that was subnormal. Concurrent with these six abrupt changes in TSH response, the mean serum T(3) level increased only from 105+/-5 to 129+/-9 ng/100 ml, and the mean serum T(4) level increased only from 4.9+/-0.8 to 6.3+/-0.5 mug/100 ml. These data demonstrate the extreme sensitivity of TRH-induced TSH release to inhibition by the chronic administration of quantities of T(3) + T(4) which do not raise serum T(3) and T(4) levels above the normal ranges.

Adult↗

PubMed1972