Structural and functional analysis of the B cell epitopes recognized by anti-receptor autoantibodies in patients with Chagas' disease.
IgG fractions of patients were screened for autoantibodies against the beta1- and beta2-adrenoceptors and the M2 acetylcholine receptor by enzyme immunoassays and surface plasmon resonance (SPR) using peptides corresponding to the second extracellular loop of these receptors. A high prevalence of anti-M2 acetylcholine receptor and, in decreasing order, of anti-beta1- and anti-beta2-adrenoceptor autoantibodies was shown. The enzyme immunoassays and the SPR studies on the anti-beta1 adrenoceptor and the M2 acetylcholine receptor autoantibodies were dependent on the ionic strength of the interaction buffer, suggesting the importance of electrostatic interactions in Ab recognition. IgG fractions showed chronotropic effects on neonatal rat cardiomyocytes in vitro. The positive chronotropic effect was enhanced in the presence of 1 microM of atropine, demonstrating a muscarinic stimulation by the IgG fractions in the presence of a beta-adrenergic stimulation, which was blocked by the use of 1 microM of the beta1-selective antagonist bisoprolol. The beta2-selective antagonist ICI 118,551 only partially inhibited the positive chronotropic effect induced by the IgG fractions, confirming the minor functional importance of autoantibodies against the beta2-adrenoceptor. Affinity-purified Abs confirmed that Abs against the beta1-adrenoceptors and the M2 muscarinic receptors exist together with an Ab population recognizing a cross-reactive epitope on both receptors. This epitope could be identified as a polyanionic stretch present in the second extracellular loop of both the beta1-adrenoceptor and the M2 acetylcholine receptor. This stretch corresponds to the previously determined cross-reactive epitope between the P0 ribosomal protein of Trypanosoma cruzi and the beta1-adrenoceptor.
Elies
R
R
CJF INSERM Unit 93/09 Immunology for Infectious Diseases, Pharmaceutical Sciences, University Francois Rabelais, Tours, France.
Ferrari
I
I
Wallukat
G
G
Lebesgue
D
D
Chiale
P
P
Elizari
M
M
Rosenbaum
M
M
Hoebeke
J
J
Levin
M J
MJ
D000319:Adrenergic beta-Antagonists / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology*; D001327:Autoimmune Diseases / Q000276:immunology*; D017298:Bisoprolol / Q000494:pharmacology; D002598:Chagas Cardiomyopathy / Q000276:immunology; D014355:Chagas Disease / Q000276:immunology*; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D008969:Molecular Sequence Data; D009206:Myocardium / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D011412:Propanolamines / Q000494:pharmacology; D051381:Rats; D018342:Receptors, Adrenergic, beta-1 / Q000737:chemistry / Q000276:immunology*; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry / Q000276:immunology*; D011976:Receptors, Muscarinic / Q000737:chemistry / Q000276:immunology*
D000319:Adrenergic beta-Antagonists; D001323:Autoantibodies; D001324:Autoantigens; D000939:Epitopes; D007074:Immunoglobulin G; D010446:Peptide Fragments; D011412:Propanolamines; D018342:Receptors, Adrenergic, beta-1; D018343:Receptors, Adrenergic, beta-2; D011976:Receptors, Muscarinic; C026777:ICI 118551; D017298:Bisoprolol