Prevention of passively transferred experimental autoimmune myasthenia gravis by a phage library-derived cyclic peptide.
Many pathogenic antibodies in myasthenia gravis (MG) and its animal model, experimental autoimmune MG (EAMG), are directed against the main immunogenic region (MIR) of the acetylcholine receptor (AcChoR). These antibodies are highly conformation dependent; hence, linear peptides derived from native receptor sequences are poor candidates for their immunoneutralization. We employed a phage-epitope library to identify peptide-mimotopes capable of preventing the pathogenicity of the anti-MIR mAb 198. We identified a 15-mer peptide (PMTLPENYFSERPYH) that binds specifically to mAb 198 and inhibits its binding to AcChoR. A 10-fold increase in the affinity of this peptide was achieved by incorporating flanking amino acid residues from the coat protein as present in the original phage library. This extended peptide (AEPMTLPENYFSERPYHPPPP) was constrained by the addition of cysteine residues on both ends of the peptide, thus generating a cyclic peptide that inhibited the binding of mAb 198 to AcChoR with a potency that is three orders of magnitude higher when compared with the parent library peptide. This cyclic peptide inhibited the in vitro binding of mAb 198 to AcChoR and prevented the antigenic modulation of AcChoR caused by mAb 198 in human muscle cell cultures. The cyclic peptide also reacted with several other anti-MIR mAbs and the sera of EAMG rats. In addition, this peptide blocked the ability of mAb 198 to passively transfer EAMG in rats. Further derivatization of the cyclic peptide may aid in the design of suitable synthetic mimotopes for modulation of MG.
Venkatesh
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N
Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel.
Im
S H
SH
Balass
M
M
Fuchs
S
S
Katchalski-Katzir
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology; D000918:Antibody Specificity; D001435:Bacteriophages; D001667:Binding, Competitive; D000939:Epitopes / Q000008:administration & dosage / Q000276:immunology; D005260:Female; D006801:Humans; D007106:Immune Sera / Q000276:immunology / Q000378:metabolism; D008969:Molecular Sequence Data; D009157:Myasthenia Gravis / Q000276:immunology / Q000517:prevention & control*; D009842:Oligopeptides / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology; D019151:Peptide Library; D010456:Peptides, Cyclic / Q000008:administration & dosage* / Q000737:chemistry / Q000276:immunology; D011485:Protein Binding; D051381:Rats; D011917:Rats, Inbred Lew; D011950:Receptors, Cholinergic / Q000276:immunology / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000939:Epitopes; D007106:Immune Sera; D009842:Oligopeptides; D019151:Peptide Library; D010456:Peptides, Cyclic; D011950:Receptors, Cholinergic