Open AccessRhabdomyosarcoma cell line can be used for the isolation of soluble acetylcholine receptor and for assaying blocking and modulating autoantibodies
Author(s) -
Wu James T.,
Astill Mark,
Lloyd Christopher,
Salmon V. C.
Publication year - 1993
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.1860070104
Subject(s) - acetylcholine receptor , myasthenia gravis , autoantibody , cell culture , antibody , receptor , chemistry , microbiology and biotechnology , rhabdomyosarcoma , blocking (statistics) , biology , immunology , biochemistry , pathology , medicine , sarcoma , genetics , statistics , mathematics
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 (αButx) in a time‐dependent and saturable fashion. The apparent dissociation constant (5.3 × 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 125 l‐αButx. Soluble AChR from RD cells can be labeled specifically with 125 l‐αButx 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 αButx. 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. © 1993 Wiley‐Liss, Inc.