title: Expression of nicotinic acetylcholine receptor subunits in HEp-2 cells for immunodetection of autoantibody specificities in sera from Myasthenia gravis patients.
abstract: Myasthenia gravis (MG) is an autoimmune disease characterized by the formation of pathogenic autoantibodies mostly targeting the nicotinic acetylcholine receptor (AChR). The AChR is composed of two alpha subunits and one subunit of each beta, delta and gamma (fetal AChR), or epsilon (adult AChR), respectively. Serological diagnostics is commonly done by radioimmunoassay (RIA). Here we used an indirect immunofluorescence assay with MG patient sera on transiently transfected HEp-2 cells expressing selected components of the AChR. Our data show that already 12 out of 13 MG patient sera showed autoantibody binding to HEp-2 cells transfected to express the alpha subunit solely. Interestingly, 11 out of 13 patient sera reacted positive with cells transfected to reconstitute the complete fetal AChR, but only 6 out of 13 sera showed positive signals with cells expressing the components of adult AChR. Moreover, there was no strict correlation of the serum concentration required to obtain clear-cut fluorescence signals to the antibody titer as measured by RIA. It will be an interesting topic to further investigate if the optimal serum dilution for indirect immunofluorescence as well as the autoantibody binding preferences to defined AChR subunits and to the adult versus the fetal receptor variant could provide additional predictive value in MG diagnostics.
authors.affiliation: Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, Germany.|Institute of Immunology, Medical Faculty of the Technical University of Dresden, Germany.
authors.forename: S
authors.initials: S
authors.lastname: George
authors.affiliation: Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Noack
authors.affiliation: Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Vanek
authors.affiliation: Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Rentzsch
authors.affiliation: Institute of Immunology, Medical Faculty of the Technical University of Dresden, Germany.
authors.forename: N
authors.initials: N
authors.lastname: Röber
authors.affiliation: Institute of Immunology, Medical Faculty of the Technical University of Dresden, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Conrad
authors.affiliation: Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, Germany.|Generic Assays GmbH, Dahlewitz, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Roggenbuck
authors.affiliation: Faculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, Germany.
authors.forename: J-H
authors.initials: JH
authors.lastname: Küpper
mesh_terms: D001323:Autoantibodies / Q000276:immunology*; D002478:Cells, Cultured; D006801:Humans; D007822:Laryngeal Neoplasms / Q000276:immunology* / Q000473:pathology; D009157:Myasthenia Gravis / Q000097:blood / Q000276:immunology*; D011978:Receptors, Nicotinic / Q000096:biosynthesis*
chemical_list: D001323:Autoantibodies; D011978:Receptors, Nicotinic
keywords: Acetylcholine receptor; HEp-2 cells; Myasthenia gravis; autoantibodies; cell-based assay; indirect immunofluorescence; multiplex diagnostics; radioimmunoassay; serological diagnostics