A new chemiluminescent assay for the rapid detection of thyroid stimulating antibodies in Graves' disease.
OBJECTIVE
Thyroid stimulating antibodies (TSAb) are the cause of Graves' disease (GD). At present, these antibodies can only be measured using bioassays, which are generally time consuming and difficult to apply to whole serum. Here we describe a new chemiluminescent assay for the detection of TSAb in serum samples.


METHODS
A Chinese hamster ovary (CHO) cell line, stably transformed with a reporter plasmid containing the firefly luciferase gene under the transcriptional control of multiple cAMP responsive elements (CRE), was transfected with the human thyroid stimulating hormone (TSH) receptor. A clonal cell line, (NA-4), was obtained which showed a dose-dependent increase in luciferase activity in response to bovine and human TSH stimulation. NA-4 was subsequently used to study TSAb activity in 42 GD sera.


RESULTS
Of the GD sera 25 (60%) produced an increase in luciferase activity of 1.4-9 fold that obtained with control sera. Results were compared with an established bioassay for TSAb, based on the direct measurement of intracellular cAMP, and there was a significant correlation between the two assays (r = 0.8; P < 0.0001). IgG from GD patients, but not controls, also increased luciferase activity in NA-4 cells using concentrations as low as 3 mg/l in selected samples.


CONCLUSIONS
The present report describes a simple, sensitive and rapid assay for the detection of TSAb, with application both in the clinical analysis of GD patient sera and as a potential research tool for use in the isolation of TSAb monoclonal antibodies.
Watson
P F
PF
Division of Clinical Sciences Centre, University of Sheffield, Northern General Hospital, UK. p.f.watson@sheffield.ac.uk
Ajjan
R A
RA
Phipps
J
J
Metcalfe
R
R
Weetman
A P
AP
D000818:Animals; D016466:CHO Cells; D002417:Cattle; D006224:Cricetinae; D006111:Graves Disease / Q000175:diagnosis* / Q000276:immunology; D006801:Humans; D018828:Immunoglobulins, Thyroid-Stimulating / Q000032:analysis*; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements
D018828:Immunoglobulins, Thyroid-Stimulating; D008156:Luciferases