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Abstract:

High levels of antibodies against the C-terminus of the Trypanosoma cruzi TcP2β ribosomal protein, defined by the peptide EEEDDDMGFGLFD, named R13, have been measured in sera from patients with chronic Chagas' Heart Disease (cChHD). These antibodies also recognize an epitope on the second extracellular loop of the β1-adrenergic receptor, inducing a functional response on cardiomyocytes. The aim of this study was to gain novel insights into the structural basis of this cross-reactivity as well as to evaluate the origin of anti-M2- cholinergic receptor antibodies, which are also commonly found in cChHD patients. To address these questions we immunopurified anti-R13 antibodies and studied the structural requirements of epitope recognition. Results showed that the immunopurified antibodies recognized a conformation of R13 in which the third Glu residue was essential for binding, explaining their low affinity for the mammalian homologue (peptide H13: EESDDDMGFGLFD). Alanine mutation scanning showed individual variations in epitope recognition in each of the studied patients. The importance of a negatively charged residue at position 3 for the recognition of anti-R13 antibodies was further confirmed by competition experiments using a Ser3-phosphorylated H13 analogue, which had 10 times more affinity for the anti-R13 antibody than the native H13 peptide. Moreover, anti-R13 antibodies stimulated either the β1-adrenergic or the M2-cholinergic receptor, in strict agreement with the functional properties of the IgG fractions from which they derived, demonstrating that the same parasite antigen may generate antibody specificities with different functional properties. This may be a clue to explain the high variability of electrophysiological disturbances found in cChHD.

Registro:

Documento: Artículo
Título:Structural and functional complexity of the humoral response against the Trypanosoma cruzi ribosomal P2β protein in patients with chronic Chagas' heart disease
Autor:Mahler, E.; Hoebeke, J.; Levin, M.J.
Filiación:Depto. Fisiol. y Biol. Molec. y Cel., Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina
Lab. Biol. Molec. Enfermedad Chagas, Inst. Invest. Ing. Genet. y Biol. M., Argentina
UPR 9021 of the C.N.R.S, Inst. de Biol. Molec. et Cellulaire, Strasbourg, France
Lab. of Molec. Biol. of Chagas' Dis., Inst. Genetic Eng. and Molec. Biol., Vta. de Obligado 2490, 1428-Capital Federal, Argentina
Palabras clave:β1-adrenergic receptor; Chronic Chagas' heart disease; M2-cholinergic receptor; Ribosomal P proteins; Trypanosoma cruzi; beta 1 adrenergic receptor; epitope; glutamylglutamylserylaspartylaspartylaspartylmethionylglycylphenylalanylgly cylleucylphenylalanylaspartic acid; muscarinic M2 receptor; parasite antigen; peptide antibody; peptide derivative; protozoal protein; r13 antibody; ribosome protein; trypanosoma cruzi ribosomal p2beta protein; unclassified drug; animal cell; animal tissue; antibody affinity; antibody detection; antibody specificity; antigen recognition; article; Chagas disease; chronic disease; controlled study; cross reaction; host parasite interaction; human; humoral immunity; immunoreactivity; major clinical study; molecular recognition; nonhuman; parasite virulence; pathophysiology; priority journal; protein binding; protein phosphorylation; protozoal infection; rat; structure activity relation; Trypanosoma cruzi; Animals; Antibodies, Protozoan; Cells, Cultured; Chagas Disease; Cross Reactions; Enzyme-Linked Immunosorbent Assay; Epitope Mapping; Epitopes; Humans; Myocytes, Cardiac; Rats; Rats, Sprague-Dawley; Receptors, Adrenergic, beta; Receptors, Cholinergic; Ribosomal Proteins; Trypanosoma cruzi
Año:2004
Volumen:136
Número:3
Página de inicio:527
Página de fin:534
DOI: http://dx.doi.org/10.1111/j.1365-2249.2004.02480.x
Título revista:Clinical and Experimental Immunology
Título revista abreviado:Clin. Exp. Immunol.
ISSN:00099104
CODEN:CEXIA
CAS:Antibodies, Protozoan; Epitopes; Receptors, Adrenergic, beta; Receptors, Cholinergic; Ribosomal Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00099104_v136_n3_p527_Mahler

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Citas:

---------- APA ----------
Mahler, E., Hoebeke, J. & Levin, M.J. (2004) . Structural and functional complexity of the humoral response against the Trypanosoma cruzi ribosomal P2β protein in patients with chronic Chagas' heart disease. Clinical and Experimental Immunology, 136(3), 527-534.
http://dx.doi.org/10.1111/j.1365-2249.2004.02480.x
---------- CHICAGO ----------
Mahler, E., Hoebeke, J., Levin, M.J. "Structural and functional complexity of the humoral response against the Trypanosoma cruzi ribosomal P2β protein in patients with chronic Chagas' heart disease" . Clinical and Experimental Immunology 136, no. 3 (2004) : 527-534.
http://dx.doi.org/10.1111/j.1365-2249.2004.02480.x
---------- MLA ----------
Mahler, E., Hoebeke, J., Levin, M.J. "Structural and functional complexity of the humoral response against the Trypanosoma cruzi ribosomal P2β protein in patients with chronic Chagas' heart disease" . Clinical and Experimental Immunology, vol. 136, no. 3, 2004, pp. 527-534.
http://dx.doi.org/10.1111/j.1365-2249.2004.02480.x
---------- VANCOUVER ----------
Mahler, E., Hoebeke, J., Levin, M.J. Structural and functional complexity of the humoral response against the Trypanosoma cruzi ribosomal P2β protein in patients with chronic Chagas' heart disease. Clin. Exp. Immunol. 2004;136(3):527-534.
http://dx.doi.org/10.1111/j.1365-2249.2004.02480.x