Rhabdomyosarcoma cell line can be used for the isolation of soluble acetylcholine receptor and for assaying blocking and modulating autoantibodies.
We found that the Rhabdomyosarcoma (RD) cell line expresses human acetylcholine receptor (AChR) based on the following evidences: 1. Soluble AChR can be isolated from RD cells following the isolation procedure for AChR from human muscle; 2. Intact RD cells bind to alpha-bungarotoxin (alpha Butx) in a time-dependent and saturable fashion. The apparent dissociation constant (5.3 x 10(-10) M) is very similar to that reported for TE671 cells, which is known to express AChR; 3. Like fresh muscle culture, RD cells not only bind but also internalize 125I-alpha Butx. Soluble AChR from RD cells can be labeled specifically with 125I-alpha Butx and then used to quantify binding autoantibodies in myasthenic patients. We also demonstrate that blocking antibodies can be detected in sera from patients with myasthenia gravis (MG) using RD cells and the ability of RD cells to internalize alpha Butx. Consequently, RD cells can be used as a reliable source for obtaining soluble AChR and as a replacement for rodent or human muscle cultures in measuring blocking and modulating antibodies.
Wu
J T
JT
Department of Pathology, University of Utah Medical Center, Salt Lake City.
Astill
M
M
Lloyd
C
C
Salmon
V C
VC
D001323:Autoantibodies / Q000032:analysis*; D001667:Binding, Competitive; D002038:Bungarotoxins / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D009157:Myasthenia Gravis / Q000276:immunology; D011950:Receptors, Cholinergic / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D012208:Rhabdomyosarcoma / Q000276:immunology* / Q000378:metabolism*; D012995:Solubility; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000378:metabolism
D001323:Autoantibodies; D002038:Bungarotoxins; D011950:Receptors, Cholinergic