title: Human anti-nicotinic acetylcholine receptor recombinant Fab fragments isolated from thymus-derived phage display libraries from myasthenia gravis patients reflect predominant specificities in serum and block the action of pathogenic serum antibodies.
abstract: Myasthenia gravis (MG) is a prototype Ab-mediated autoimmune disease in which Abs against nicotinic acetylcholine receptors (AChR) induce loss of functional receptors at the neuromuscular junction. Germinal centers present in MG hyperplastic thymus contain activated B cells spontaneously producing anti-human AChR (anti-huAChR) Ab in vitro. To access the anti-huAChR repertoire, phage display Fab libraries of thymic lymphocytes were constructed from two MG patients. Four Fabs highly specific for huAChR were isolated that bind to determinants in or near the main immunogenic region. These anti-huAChR Fabs showed evidence of significant somatic mutations, supporting the idea that the anti-huAChR Ab response in MG patients is driven by Ag. Two Fabs were able to inhibit up to 90% of donor serum anti-huAChR Abs. Competition with serum anti-huAChR Ab was also observed in unrelated MG patients and indicate that anti-huAChR Fabs bind to epitopes on huAChR recognized by the majority of MG patients. In vitro antigenic modulation studies demonstrated that anti-huAChR Fabs were able to induce AChR loss when cross-linked by an anti-Fab Ab but not as monovalent Fab. Moreover, anti-huAChR Fabs were able to protect against AChR loss by antigenic modulation induced by MG serum Abs, suggesting a potential therapeutic role for these recombinant Fabs in patients with a myasthenic crisis.
authors.affiliation: The Scripps Research Institute, Department of Immunology, La Jolla, CA 92037, USA.
authors.forename: Y F
authors.initials: YF
authors.lastname: Graus
authors.forename: M H
authors.initials: MH
authors.lastname: de Baets
authors.forename: P W
authors.initials: PW
authors.lastname: Parren
authors.forename: S
authors.initials: S
authors.lastname: Berrih-Aknin
authors.forename: J
authors.initials: J
authors.lastname: Wokke
authors.forename: P J
authors.initials: PJ
authors.lastname: van Breda Vriesman
authors.forename: D R
authors.initials: DR
authors.lastname: Burton
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D019138:Antibodies, Blocking / Q000494:pharmacology* / Q000627:therapeutic use; D000918:Antibody Specificity; D001323:Autoantibodies / Q000009:adverse effects* / Q000097:blood; D001324:Autoantigens / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive / Q000276:immunology; D003090:Coliphages / Q000235:genetics; D003432:Cross-Linking Reagents; D018604:Epitope Mapping; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000737:chemistry*; D007140:Immunoglobulin Fab Fragments / Q000235:genetics* / Q000302:isolation & purification / Q000627:therapeutic use; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009157:Myasthenia Gravis / Q000209:etiology* / Q000235:genetics / Q000276:immunology*; D011978:Receptors, Nicotinic / Q000276:immunology* / Q000378:metabolism; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000494:pharmacology / Q000627:therapeutic use; D017421:Sequence Analysis; D013950:Thymus Gland / Q000821:virology
chemical_list: D019138:Antibodies, Blocking; D001323:Autoantibodies; D001324:Autoantigens; D003432:Cross-Linking Reagents; D007106:Immune Sera; D007140:Immunoglobulin Fab Fragments; D011978:Receptors, Nicotinic; D011994:Recombinant Proteins