Development of an assay for modulating anti-acetylcholine receptor autoantibodies using human rhabdomyosarcoma cell line.
Three types of autoantibodies against the acetylcholine receptors (AChR) of skeletal muscle are detectable in patients with myasthenia gravis including binding, blocking, and modulating anti-AChR antibodies. Modulating autoantibodies correlate best with the severity of the disease, but are also technically most difficult to measure because the assay generally requires fresh human muscle cells. We have developed an assay for the modulation of anti-AChR antibodies using a rhabdomyosarcoma (RD) cell line expressing AChR on the cell surface. By decreasing the FetalClone III serum from 10% to 0.5% in Eagles Minimal Essential Medium (EMEM) we were able to increase the number of AChR on RD cells to meet the need of sensitivity of the assay. The extent of modulation was determined as the percent of AChR internalized in the presence or absence of modulating autoantibodies. Less than 6% modulation was found with the normal serum (n = 42). The CVs of both the intra- and day-to-day precision were less than 20%. When clinical samples (n = 105) were assayed in our laboratory and also at Nichols Institute, a correlation coefficient of 0.816 was obtained. The selection of RD cell line, the success of increasing the expression of the AChR on RD cells and the use of 125I alpha-bungarotoxin of high specific activity allowed the establishment of an assay which can be used in routine clinical laboratory for the measurement of modulating anti-AChR autoantibodies for the management of patients with myasthenia gravis.
Lyons
B W
BW
Department of Pathology and Associated Regional University Pathologists, University of Utah Health Science Complex, Salt Lake City 84132, USA.
Wu
L L
LL
Astill
M E
ME
Wu
J T
JT
D001323:Autoantibodies / Q000097:blood*; D002038:Bungarotoxins; D006801:Humans; D009157:Myasthenia Gravis / Q000097:blood / Q000276:immunology*; D011950:Receptors, Cholinergic / Q000276:immunology* / Q000302:isolation & purification; D012016:Reference Values; D012208:Rhabdomyosarcoma; D014407:Tumor Cells, Cultured
D001323:Autoantibodies; D002038:Bungarotoxins; D011950:Receptors, Cholinergic