title: Inhibition of alpha-bungarotoxin binding to acetylcholine receptors by antisera from animals with experimental autoimmune myasthenia gravis.
abstract: Conditions are described for an assay that allows the percent inhibition of alpha-bungarotoxin binding to acetylcholine receptors by antisera and monovalent antigen-binding fragments of antibody molecules (Fab) to be determined. Anti-Torpedo californica acetylcholine-receptor antisera, prepared in New Zealand White rabbits and Lewis rats, were tested for the ability to inhibit [125I]-alpha-bungarotoxin binding to membrane-associated and detergent-solubilized T californica acetylcholine receptors. Similar inhibition studies were performed using rabbit antisera and antigen-binding fragments prepared against each of the four acetylcholine receptor subunits. Antisera and antigen-binding fragments prepared against intact receptor could inhibit a maximum of 50% of the alpha-bungarotoxin binding to solubilized receptor. The results using monovalent antigen-binding fragments indicated that the inhibition was not due to antibody-mediated aggregation of receptor molecules. Rabbits and rats immunized with receptor denatured by sodium dodecyl sulfate all produced antisera that could bind to nondenatured receptor, but none of these animals developed experimental autoimmune myasthenia gravis. These results suggest that the antigenic determinants present on acetylcholine receptors responsible for induction of experimental autoimmune myasthenia gravis are lost with sodium dodecyl sulfate denaturation. A strong correlation was also observed between the presence of experimental autoimmune myasthenia gravis in rats and rabbits and the ability of the antisera from these animals to inhibit 50% of alpha-bungarotoxin binding to solubilized acetylcholine receptors.
mesh_terms: D000818:Animals; D001327:Autoimmune Diseases / Q000276:immunology*; D002038:Bungarotoxins / Q000378:metabolism*; D004195:Disease Models, Animal; D005399:Fishes; D007106:Immune Sera; D007118:Immunoassay; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007700:Kinetics; D009157:Myasthenia Gravis / Q000276:immunology*; D011817:Rabbits / Q000276:immunology; D051381:Rats; D011917:Rats, Inbred Lew / Q000276:immunology; D011950:Receptors, Cholinergic / Q000378:metabolism*
chemical_list: D002038:Bungarotoxins; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D011950:Receptors, Cholinergic