Binding of peptides of the human acetylcholine receptor alpha-subunit to HLA class II of patients with myasthenia gravis.
MG is an autoimmune disease in which T cells specific to T-cell epitopes of the human acetylcholine receptor play a role. We have identified two peptides, p195-212 and p259-271, of the human acetylcholine receptor alpha-subunit, to which PBLs of MG patients responded by proliferation. Nevertheless, proliferation assays are relatively complicated to perform and might be affected by medications taken by the patients. Therefore, we tested the possibility of using a different assay to determine recognition of these peptides by MG patients. Thus, we performed a direct binding assay using biotinylated peptides and APCs from peripheral blood of MG patients and healthy controls. With this assay we detected the binding of the two peptides to the surface of intact APCs of both MG patients and control donors. Moreover, the presentation of peptide p259-271 by individuals with MG was significantly higher than that observed in healthy subjects. The peptides were specifically bound to HLA class II determinants on the APCs, as shown by inhibition with antibodies to the HLA class II haplotypes of the individuals investigated. Moreover, the binding of these peptides was in correlation with their ability to induce specific proliferative responses of peripheral blood T cells of these patients. The ability to screen for potentially pathogenic epitopes in each patient is of importance for the future design of specific inhibitory analogues that might be used to treat MG.
Zisman
E
E
Department of Chemical Immunology, Weizmann Institute of Science, Rehovot, Israel.
Brautbar
C
C
Sela
M
M
Abramsky
O
O
Battat
S
S
Kirshner
S L
SL
Katz-Levy
Y
Y
Dayan
M
M
Mozes
E
E
D000595:Amino Acid Sequence; D000938:Antigen-Presenting Cells / Q000378:metabolism; D001710:Biotin / Q000378:metabolism; D005260:Female; D006681:HLA-D Antigens / Q000378:metabolism*; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D009157:Myasthenia Gravis / Q000276:immunology / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology*; D011485:Protein Binding / Q000276:immunology; D011950:Receptors, Cholinergic / Q000737:chemistry / Q000378:metabolism*
D006681:HLA-D Antigens; D010446:Peptide Fragments; D011950:Receptors, Cholinergic; D001710:Biotin