Participation of nitric oxide signaling system in the cardiac muscarinic cholinergic effect of human chagasic IgG.
The possible role of altered humoral immune response in the pathogenesis of the chronic chagasic cardioneuromyopathy was examined by analyzing the interaction of IgG from T. cruzi infected patients with cardiac muscarinic acetylcholine receptors (mAChR). Human chagasic IgG by activating cardiac M2 mAChR, simulated the agonist actions triggering negative inotropic effect, inositol phosphate accumulation, nitric oxide synthase stimulation and increased production of cyclic GMP. Inhibitors of phospholipase C, protein kinase C, calcium/calmodulin, nitric oxide synthase and guanylate cyclase activities; prevented chagasic IgG effects on signaling pathways involved in M2 mAChR activation. In addition, sodium nitroprusside or 8-bromo cyclic GMP, mimicked the chagasic IgG effect associated with cholinergic-mediated cellular transmembrane signals. Moreover, these chagasic IgG immunoprecipitated the mAChRs solubilized from cardiac membranes. By means of SDS-PAGE and immunoblotting analysis, chagasic sera recognized a band of 70-75 kDa. The major protein recognized by chagasic IgG had an Rf coincident with the peak of [3H] propylbenzilylcholine mustard with an apparent molecular weight similar to that of mAChRs, which disappeared in the presence of atropine. The specificity of this interaction was checked by immunoprecipitation of rat cardiac mAChR and immunoblotting of pure human M2 mAChRs. Chronic interaction of chagasic IgG with myocardial mAChRs, behaving as a muscarinic agonist, might lead to cell dysfunction or tissue damage. Also, these antibodies could produce desensitization, internalization or degradation of mAChRs; explaining the progressive blockade of mAChRs in myocardium with parasympathetic denervation, a phenomenon that has been described in the course of Chagas' cardioneuromyopathy.
Sterin-Borda
L
L
Centro de Estudios Farmacológicos y Botánicos (CEFYBO), Universidad de Buenos Aires, Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Buenos Aires, Argentina.
Leiros
C P
CP
Goin
J C
JC
Cremaschi
G
G
Genaro
A
A
Echagüe
A V
AV
Borda
E
E
D000328:Adult; D000368:Aged; D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D015153:Blotting, Western; D014355:Chagas Disease / Q000503:physiopathology*; D006152:Cyclic GMP / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007295:Inositol Phosphates / Q000378:metabolism; D008875:Middle Aged; D009200:Myocardial Contraction; D009569:Nitric Oxide / Q000502:physiology*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D051381:Rats; D011976:Receptors, Muscarinic / Q000276:immunology / Q000502:physiology*; D015398:Signal Transduction
D000913:Antibodies, Protozoan; D007074:Immunoglobulin G; D007295:Inositol Phosphates; D011976:Receptors, Muscarinic; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D006152:Cyclic GMP