Isolation and functional characterization of anti-acetylcholine receptor subunit-specific autoantibodies from myasthenic patients: receptor loss in cell culture.
The muscle nicotinic acetylcholine receptor (nAChR) is the major autoantigen in the autoimmune disease myasthenia gravis (MG), in which autoantibodies bind to, and cause loss of, nAChRs. Antibody-mediated nAChR loss is caused by the action of complement and by the acceleration of nAChR internalization caused by antibody-induced cross-linking of nAChR molecules (antigenic modulation). To obtain an insight into the role of the various anti-nAChR antibody specificities in MG, we have studied nAChR antigenic modulation caused by isolated anti-subunit autoantibodies. Autoantibodies against the nAChR alpha or beta subunits were isolated from four MG sera by affinity chromatography on columns carrying immobilized recombinant extracellular domains of human nAChR expressed in the yeast Pichia pastoris. The isolated anti-alpha and anti-beta autoantibodies, as well as untreated MG sera, induced nAChR antigenic modulation in TE671 cells. Partially antibody-depleted sera exhibited reduced modulating activity, whereas a serum completely depleted of anti-nAChR antibodies exhibited no nAChR modulation. Interestingly, the anti-alpha autoantibodies were, on average, approximately 4.3 times more effective than the anti-beta autoantibodies. The present work supports the notion that anti-nAChR autoantibodies may be the sole nAChR-reducing factor in anti-nAChR antibody-seropositive MG, and that anti-alpha-subunit autoantibodies are the dominant pathogenic autoantibody specificity.
Sideris
Sotiris
S
Department of Pharmacy, University of Patras, 26504 Patras, Greece.
Lagoumintzis
George
G
Kordas
Gregory
G
Kostelidou
Kalliopi
K
Sotiriadis
Alexandros
A
Poulas
Konstantinos
K
Tzartos
Socrates J
SJ
D000918:Antibody Specificity; D001323:Autoantibodies / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D001324:Autoantigens / Q000502:physiology; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007163:Immunosorbent Techniques; D008297:Male; D009157:Myasthenia Gravis / Q000097:blood* / Q000276:immunology*; D021122:Protein Subunits / Q000276:immunology; D011950:Receptors, Cholinergic / Q000145:classification / Q000172:deficiency / Q000276:immunology*; D012208:Rhabdomyosarcoma / Q000473:pathology
D001323:Autoantibodies; D001324:Autoantigens; D021122:Protein Subunits; D011950:Receptors, Cholinergic