Exp. Clin. Endocrino!, loo (1992) 45-47
Experimental and Clinical Endocrinology © 1992 Johann Ambrosius Barth
H. Gerber, H. J. Peter and H. Studer Laboratories of Experimental Endocrinology, Department of Internal Medicine, University of Berne, Inseispital, Berne, Switzerland
Key words: Thyroid - Graves' disease - Nude mouse - TSH receptor antibodies - Species specificity
webe stimulieren. Diese Resultate belegen klar einen speziesspezifischen Effekt von TSI.
Zusammenfassung: Die Xenotransplantation von humanem
Summary: Administration of sera from patients with active
Schilddrüsengewebe auf Nacktmäuse bietet die Möglichkeit, die Wirkung von Thyreoidea-stimulierenden Immunglobulinen (TSI) auf das Schilddrüsengewebe verschiedener Spezies vergleichend zu studieren. In der vorliegenden Arbeit zeigen wir, daß von 8 Basedow-Seren nur eines die Radiojodaufnahme der Mäuseschilddrüsen stimuliert, während alle 8 Seren die Radiojodaufnahme im xenotransplantierten humanen Schilddrüsenge-
Graves' disease to nude mice bearing human thyroid xenografts provides the opportunity to compare directly the effects of TSAb on thyroid tissue of different species. The results presented in this paper, namely the fact that only one of 8 Graves' sera increased the RAIU of the nu/nu mouse thyroids modestly while all sera increased the RAIU in the transplanted human tissue considerable, clearly show a species specific effect of TSAb.
Introduction
by Rapoport et al. (1982), Yamashita et al. (1986), De Bruin (1988), and by our group (Gerber et al., 1986,
The pathogenetic role of thyroid-stimulating antibodies (TSAb) causing follicular cell hyperfunction in Graves' disease is firmly established (McKenzie und Zakarija, 1991; Rees Smith et al., 1988; Volpé, 1986; Zakarija and McKenzie, 1991). However, TSAb are not detected by currently used test systems in some untreated hyperthyroid Graves' patients, and in the individual patient test results for TSAb obtained with different assays can vary widely.
1987).
In the present study, we have investigated the effect of
serum from patients with active Graves' disease on radioiodine uptake (RAIU) of the thyroid gland of nude mice and, for comparison, of xenotransplanted human thyroid tissue.
For these discrepancies between TSAb measured and
Material and Methods
presence or severity of thyrotoxicosis and those between different TSAb assays, several explanations have been proposed in the literature (McKenzie and Zakarija, 1991;
In 6 experiments, human thyroid tissue fragments obtained at
Rees Smith et al., 1988; Volpé, 1986; Zakarija and McKenzie, 1991). One explanation for some of the discrepancies between laboratory test and clinical picture might be variable crossreactivity of TSAb in different mammalian tissue. Indeed, evidence for species specificity in the interaction between TSAb and their receptor has been presented many years ago by Zakarija and McKenzie (1978 a, 1978 b), later also
surgery (from 3 Graves' goiters, 2 normal glands and 1 euthyroid nodular goiter) were transplanted onto dysthymic nude nu/nu ICR mice as described in detail elsewhere (Peter et al., 1985, 1987). The animals were kept under pathogen free conditions in a 12 h light 12h dark cycle and fed with cereal based mouse breeding diet (Nafag AG, Gossau, Switzerland) sterilized by irradiation with
2.5 megarads. 5 weeks after transplantation, L-thyroxine (T4) 0.5 tg/ml (Fluka AG, Buchs, Switzerland) was added to the drinking water in order to suppress the mouse's own TSH. After 2
additional weeks, the animals were divided into groups of 3 to 7 animals and treated for 6 days daily i.p. with either
Downloaded by: Collections and Technical Services Department. Copyrighted material.
Effect of Human Thyroid Stimulating Autoantibodies on the Radioiodine Uptake of the Mouse Thyroid Gland
46
Exp. Clin. Endocrinol. loo (1992) 1/2
- 0.5 ml of serum from a healthy human donor whose TSH had been suppressed by T4-treatment (negative control).
Results
- 100 mU of bovine TSH (Ambinon, Organon Oss, NL) in
The results are summarized in Table 1 and Figure 1. As
0.5 ml NaC1 0.9 % (positive control), or - 0.5 ml of serum from patients with active Graves' disease (see Table 1).
expected, TSH increased the radioiodine uptáke (TAIU) of the mouse thyroids in all experiments as well as that of the
Table 1 The effect of control serum, Graves' serum and bovine TSH on the radioiodine uptake (in % of values obtained with control serum) of the nu/nu mouse thyroid gland. Means ± SD are given (n in brackets). Details see Material and Methods
only one of the 8 Graves' sera tested increased the RAIU of the mouse glands modestly to 200 O/ of that in the control group (Table 1), while all sera increased the RAJU in the
TSH
Experiment
Control serum
Graves'
2 3*
100 ± 16 (3) 100 ± 24 (4) 100 ± 7 (3)
4 5**
100 ± 34 (5) 100 ± 38 (5)
90 ± 38 (4) 92 ± 13 (4) 109 ± 8* (4) 143 ± 75 (4) 145 ± 37 (5) 200 ± 99° (5)
transplanted human tissue to 400-2700% of that in the control group (data not shown).
serum (4) (4) (4)
1402 ± 1333° 1304 ± 512°°
(4) (4)
100 ± 20 (7)
139 ± 132 (4) 96 ± 27 (6)
1139 ± 272°°°
(6)
* In Experiment 3, two different Graves' sera were tested in two experimental groups. The serum marked with * is identical to that used in Experiment 2.
** In Experiment 5, three different Graves' sera were tested in three experimental groups. p < 0.05 vs. control
p
Experimental and Clinical Endocrinology © 1992 Johann Ambrosius Barth
H. Gerber, H. J. Peter and H. Studer Laboratories of Experimental Endocrinology, Department of Internal Medicine, University of Berne, Inseispital, Berne, Switzerland
Key words: Thyroid - Graves' disease - Nude mouse - TSH receptor antibodies - Species specificity
webe stimulieren. Diese Resultate belegen klar einen speziesspezifischen Effekt von TSI.
Zusammenfassung: Die Xenotransplantation von humanem
Summary: Administration of sera from patients with active
Schilddrüsengewebe auf Nacktmäuse bietet die Möglichkeit, die Wirkung von Thyreoidea-stimulierenden Immunglobulinen (TSI) auf das Schilddrüsengewebe verschiedener Spezies vergleichend zu studieren. In der vorliegenden Arbeit zeigen wir, daß von 8 Basedow-Seren nur eines die Radiojodaufnahme der Mäuseschilddrüsen stimuliert, während alle 8 Seren die Radiojodaufnahme im xenotransplantierten humanen Schilddrüsenge-
Graves' disease to nude mice bearing human thyroid xenografts provides the opportunity to compare directly the effects of TSAb on thyroid tissue of different species. The results presented in this paper, namely the fact that only one of 8 Graves' sera increased the RAIU of the nu/nu mouse thyroids modestly while all sera increased the RAIU in the transplanted human tissue considerable, clearly show a species specific effect of TSAb.
Introduction
by Rapoport et al. (1982), Yamashita et al. (1986), De Bruin (1988), and by our group (Gerber et al., 1986,
The pathogenetic role of thyroid-stimulating antibodies (TSAb) causing follicular cell hyperfunction in Graves' disease is firmly established (McKenzie und Zakarija, 1991; Rees Smith et al., 1988; Volpé, 1986; Zakarija and McKenzie, 1991). However, TSAb are not detected by currently used test systems in some untreated hyperthyroid Graves' patients, and in the individual patient test results for TSAb obtained with different assays can vary widely.
1987).
In the present study, we have investigated the effect of
serum from patients with active Graves' disease on radioiodine uptake (RAIU) of the thyroid gland of nude mice and, for comparison, of xenotransplanted human thyroid tissue.
For these discrepancies between TSAb measured and
Material and Methods
presence or severity of thyrotoxicosis and those between different TSAb assays, several explanations have been proposed in the literature (McKenzie and Zakarija, 1991;
In 6 experiments, human thyroid tissue fragments obtained at
Rees Smith et al., 1988; Volpé, 1986; Zakarija and McKenzie, 1991). One explanation for some of the discrepancies between laboratory test and clinical picture might be variable crossreactivity of TSAb in different mammalian tissue. Indeed, evidence for species specificity in the interaction between TSAb and their receptor has been presented many years ago by Zakarija and McKenzie (1978 a, 1978 b), later also
surgery (from 3 Graves' goiters, 2 normal glands and 1 euthyroid nodular goiter) were transplanted onto dysthymic nude nu/nu ICR mice as described in detail elsewhere (Peter et al., 1985, 1987). The animals were kept under pathogen free conditions in a 12 h light 12h dark cycle and fed with cereal based mouse breeding diet (Nafag AG, Gossau, Switzerland) sterilized by irradiation with
2.5 megarads. 5 weeks after transplantation, L-thyroxine (T4) 0.5 tg/ml (Fluka AG, Buchs, Switzerland) was added to the drinking water in order to suppress the mouse's own TSH. After 2
additional weeks, the animals were divided into groups of 3 to 7 animals and treated for 6 days daily i.p. with either
Downloaded by: Collections and Technical Services Department. Copyrighted material.
Effect of Human Thyroid Stimulating Autoantibodies on the Radioiodine Uptake of the Mouse Thyroid Gland
46
Exp. Clin. Endocrinol. loo (1992) 1/2
- 0.5 ml of serum from a healthy human donor whose TSH had been suppressed by T4-treatment (negative control).
Results
- 100 mU of bovine TSH (Ambinon, Organon Oss, NL) in
The results are summarized in Table 1 and Figure 1. As
0.5 ml NaC1 0.9 % (positive control), or - 0.5 ml of serum from patients with active Graves' disease (see Table 1).
expected, TSH increased the radioiodine uptáke (TAIU) of the mouse thyroids in all experiments as well as that of the
Table 1 The effect of control serum, Graves' serum and bovine TSH on the radioiodine uptake (in % of values obtained with control serum) of the nu/nu mouse thyroid gland. Means ± SD are given (n in brackets). Details see Material and Methods
only one of the 8 Graves' sera tested increased the RAIU of the mouse glands modestly to 200 O/ of that in the control group (Table 1), while all sera increased the RAJU in the
TSH
Experiment
Control serum
Graves'
2 3*
100 ± 16 (3) 100 ± 24 (4) 100 ± 7 (3)
4 5**
100 ± 34 (5) 100 ± 38 (5)
90 ± 38 (4) 92 ± 13 (4) 109 ± 8* (4) 143 ± 75 (4) 145 ± 37 (5) 200 ± 99° (5)
transplanted human tissue to 400-2700% of that in the control group (data not shown).
serum (4) (4) (4)
1402 ± 1333° 1304 ± 512°°
(4) (4)
100 ± 20 (7)
139 ± 132 (4) 96 ± 27 (6)
1139 ± 272°°°
(6)
* In Experiment 3, two different Graves' sera were tested in two experimental groups. The serum marked with * is identical to that used in Experiment 2.
** In Experiment 5, three different Graves' sera were tested in three experimental groups. p < 0.05 vs. control
p
