Antibodies to TSH-receptor in thyroid autoimmune disease interact with monoclonal antibodies whose epitopes are broadly distributed on the receptor.

The hyperthyroidism of Graves' disease (GD) is caused by TSH-receptor (TSH-R) stimulating autoantibodies (TSAb), leading to overproduction of thyroid hormones. We present evidence for TSAb interaction with three distinct regions of the TSH-R, one in immediate vicinity of the carboxy terminal serpentine. Three murine monoclonal antibodies (MoAbs 28.

The effect of thyrotropin-receptor blocking antibodies on stimulating autoantibodies from patients with Graves' disease.

The hyperthyroidism of Graves' disease (GD) is caused by thyrotropin-receptor (TSHR) stimulating autoantibodies (TSAb), which lead to overproduction of thyroid hormones. In this study we tried to block the stimulatory effect of patients' TSAb to the TSHR with monoclonal antibodies (mAbs) and sera from hypothyroid patients. Two groups of blocking mAbs raised by different methods from two independent groups were tested for their ability to inhibit TSH binding to the TSHR, and also the binding of TSAb from the serum of patients with GD.

A coated tube assay for the detection of blocking thyrotropin receptor autoantibodies.

We developed a coated tube assay to discriminate TSH-receptor-stimulating autoantibodies [thyroid-stimulating antibodies (TSAb)] from those autoantibodies blocking TSH binding without intrinsic activation [thyroid-blocking antibodies (TBAb)]. The wild-type TSH receptor in the TSH binding-inhibitory assay was exchanged for a chimeric receptor where a TSAb epitope (amino acids 8-165) was replaced by comparable LH-R residues. Binding of (125)I-labeled TSH to this chimera could be inhibited by sera containing TBAb up to 95%.

Affinity purification and diagnostic use of TSH receptor autoantibodies from human serum.

Purification of TSH receptor autoantibodies (TRAb) from the serum of patients with Graves' disease (GD) might help to elucidate the nature of these disease causing autoantibodies. We describe here for the first time the successful affinity purification of human TRAb. Affinity purification was performed in a four step procedure with human recombinant TSH receptor (TSH-R) expressed in K562 cells.

The AMP-activated protein kinase alpha2 catalytic subunit controls whole-body insulin sensitivity.

AMP-activated protein kinase (AMPK) is viewed as a fuel sensor for glucose and lipid metabolism. To better understand the physiological role of AMPK, we generated a knockout mouse model in which the AMPKalpha2 catalytic subunit gene was inactivated. AMPKalpha2(-/-) mice presented high glucose levels in the fed period and during an oral glucose challenge associated with low insulin plasma levels.