In vivo effects of a human thyroid-stimulating monoclonal autoantibody (M22) and a human thyroid-blocking autoantibody (K1-70)

Jadwiga Furmaniak, Jane Sanders, Stuart Young, Katarzyna Kabelis, Paul Sanders, Michele Evans, Jill Clark, Jane Wilmot, Bernard Rees Smith, Jadwiga Furmaniak, Jane Sanders, Stuart Young, Katarzyna Kabelis, Paul Sanders, Michele Evans, Jill Clark, Jane Wilmot, Bernard Rees Smith

Abstract

Purpose: To study in vivo effects of the human monoclonal TSH receptor (TSHR) autoantibodies M22 (stimulating type) and K1-70 (blocking type) on thyroid hormone levels in rats.

Methods: Serum levels of total T4, free T4, M22 and K1-70 were measured following intramuscular injection of M22 IgG (2-4 μg/animal), K1-70 IgG (10-200 μg/animal) or both into rats. Thyroid pathology was assessed in M22-injected rats.

Results: Serum levels of total T4 and free T4 increased in a dose-dependent manner following injection of M22 IgG. Thyroid follicular cell hypertrophy was dependent on the dose of M22 IgG. K1-70 IgG caused a dose dependent decrease of total T4 and free T4 levels in rats receiving K1-70 only. The stimulating effects of M22 IgG on T4 levels in rats were completely inhibited by K1-70 IgG.

Conclusion: M22 is a potent stimulator of thyroid hormone secretion in vivo. In contrast, K1-70 inhibits thyroid hormone secretion in vivo. Furthermore, K1-70 has the ability to inhibit the stimulating activity of M22 in vivo and as such has potential as a new drug to block TSHR stimulation by autoantibodies in Graves' disease.

Keywords: Autoimmunity; Graves’ disease; TSH receptor; Thyroid; Thyroid-blocking antibodies; Thyroid-stimulating antibodies.

Figures

Fig. 1
Fig. 1
Injection of M22 IgG into T3-suppressed rats and serum levels of a M22 IgG, b total T4 and c free T4 (mean ± standard deviation; n = 5). Buffer only (filled diamond); 2 μg M22/rat (filled square); 4 μg M22/rat (filled triangle); 10 μg M22/rat (times symbol). All three parameters were measured in the same serum sample. The concentrations of total T4 and free T4 in all T3-suppressed rats injected with buffer only were below the detection limits of the assays, i.e. <26 nmol/L and <6.4 pmol/L, respectively
Fig. 2
Fig. 2
Histology of rat thyroid sections from a a control rat injected with buffer only showing normal thyroid histology and b a test rat at 96-h post-injection with 10 μg of M22 showing follicular cell hypertrophy characterised by a diffuse change in which the follicular epithelium showed increased height with reduced colloid
Fig. 3
Fig. 3
Injection of K1-70 IgG into rats and serum levels of a K1-70 IgG, b total T4 and c free T4 (mean ± standard deviation; n = 5). Buffer only (filled diamond); 10 μg K1-70/rat (filled square); 50 μg K1-70/rat (filled triangle); 100 μg K1-70/rat (times symbol); 200 μg K1-70/rat (filled circle). All three parameters were measured in the same serum sample. The assay detection limits for total T4 and free T4 were <26 nmol/L and <6.4 pmol/L, respectively
Fig. 4
Fig. 4
Effects of intramuscular injection of K1-70 IgG and/or M22 IgG in rats on the serum concentration of a total T4 and b free T4 (mean ± standard deviation; n = 5). Buffer only (filled diamond); 4 μg M22/rat (filled square); 200 μg K1-70/rat (filled triangle); 200 μg K1-70/rat injected 3 h  before injection of 4 μg M22/rat (times symbol); 4 μg M22/rat injected 3 h before injection of 200 μg K1-70/rat (filled circle); 200 μg K1-70 and 4 μg M22/rat injected at the same time (open circle). All three parameters were measured in the same serum sample. In the experiments when K1-70 was injected alone or in combination with M22 the total and free T4 levels were at the lower detection limits for the assays and consequently the points on the graphs are overlapping

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