Epitopes on human acetylcholine receptor defined by monoclonal antibodies and myasthenia gravis sera.
The mouse monoclonal antibody (m.ab) binding sites on human acetylcholine receptor have been mapped by inhibition by F(ab')2 m.ab fragments, by competition with a rat m.ab against the main immunogenic region (anti-m.i.r), and by their ability to protect the a-Bungarotoxin (a-BuTx) binding sites from inhibition by a myasthenia gravis (MG) plasma. Two m.abs (C3 and D6) that bind to two distinct but overlapping regions on the AChR, were inhibited by the anti-m.i.r. m.ab M35. M.abs binding to three other regions protected the a-BuTx sites by up to 50%. In further inhibition assays these m.abs were used to define the binding sites for MG anti-AChR antibodies. There was considerable heterogeneity in the antigenic specificity of the MG sera. The inhibition of MG anti-AChR by anti-m.i.r. M35 correlated highly with inhibition by mouse m.ab D6, but not with inhibition by m.ab C3. There was a correlation between inhibition by m.abs F8 and B3, although these m.abs bind to two non-overlapping regions. Some MG anti-AChR antibodies bound to epitopes that only partially overlapped those defined by m.abs. Inhibition of human antibody binding by m.abs raised against specific antigens is a useful approach that should help define the epitopes to which autoantibodies bind.
Heidenreich
F
F
Department of Neurological Science, Royal Free Hospital School of Medicine, London.
Vincent
A
A
Roberts
A
A
Newsom-Davis
J
J
D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000032:analysis*; D006801:Humans; D009157:Myasthenia Gravis / Q000276:immunology*; D011950:Receptors, Cholinergic / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D011950:Receptors, Cholinergic