An anionic synthetic sugar containing 6-SO3 -NAcGlc mimics the sulfated cruzipain epitope that plays a central role in immune recognition

• Autores: Couto, Alicia S; Soprano, Luciana L; Landoni, Malena; Pourcelot, Marilyne; Acosta, Diana M; Bultel, Laurent; Parente, Juliana E; Ferrero, Maximiliano R; Barbier, Maximilien; Dussouy, Christophe; Esteva, Monica I; Kovensky, José; Duschak, Vilma G
• Año de publicación: 2012
• Idioma: inglés
• Tipo de recurso: artículo
• Materia: Enfermedad de Chagas; Sitios de Unión de Anticuerpos; Glucosamina
• Institución: Administración Nacional de Laboratorios e Institutos de Salud "Dr. Carlos G. Malbrán"

Fil: Couto, Alicia S. Universidad de Buenos Aires. Centro de Investigaciones en Hidratos de Carbono (CIHIDECAR). Departamento de Química Orgánica; Argentina.
Fil: Soprano, Luciana L. ANLIS Dr. C.G.Malbrán. Intituto Nacional de Parasitología "Dr. Mario Fatala Chaben". Departamento de investigación; Argentina.
Fil: Landoni, Malena. Universidad de Buenos Aires. Centro de Investigaciones en Hidratos de Carbono (CIHIDECAR). Departamento de Química Orgánica; Argentina.
Fil: Pourcelot, Marilyne. Université de Picardie Jules Verne. Laboratoire des Glucides; Francia.
Fil: Acosta, Diana M. ANLIS Dr. C.G.Malbrán. Intituto Nacional de Parasitología "Dr. Mario Fatala Chaben". Departamento de investigación; Argentina.
Fil: Bultel, Laurent. Université de Picardie Jules Verne. Laboratoire des Glucides; Francia.
Fil: Parente, Juliana E. Universidad de Buenos Aires. Centro de Investigaciones en Hidratos de Carbono (CIHIDECAR). Departamento de Química Orgánica; Argentina.
Fil: Ferrero, Maximiliano R. ANLIS Dr. C.G.Malbrán. Intituto Nacional de Parasitología "Dr. Mario Fatala Chaben". Departamento de investigación; Argentina.
Fil: Barbier, Maximilien. Université de Picardie Jules Verne. Laboratoire des Glucides; Francia.
Fil: Dussouy, Christophe. Université de Picardie Jules Verne. Laboratoire des Glucides; Francia.
Fil: Esteva, Monica I. ANLIS Dr. C.G.Malbrán. Intituto Nacional de Parasitología "Dr. Mario Fatala Chaben". Departamento de investigación; Argentina.
Fil: Kovensky, José. Université de Picardie Jules Verne. Laboratoire des Glucides; Francia.
Fil: Duschak, Vilma G. ANLIS Dr. C.G.Malbrán. Intituto Nacional de Parasitología "Dr. Mario Fatala Chaben". Departamento de investigación; Argentina.
Cruzipain (Cz), the major cysteine proteinase of Trypanosoma cruzi, is a glycoprotein that contains sulfated high-mannose-type oligosaccharides. We have previously determined that these sulfate groups are targets of specific immune responses. In order to evaluate the structural requirements for antibody recognition of Cz, a systematic structure-activity study of the chemical characteristics needed for antibody binding to the Cz sulfated epitope was performed by immunoassays. With this aim, different synthesized molecules were coupled to the proteins BSA and aprotinin and confronted with (a) mouse sera specific for Cz and its carboxy-terminal (C-T) domain, (b) antibodies raised in rabbits immunized with Cz and its C-terminal domain and (c) IgGs purified from human Chagas disease sera. Our results indicate that a glucosamine containing an esterifying sulfate group in position O-6 and an N-acetyl group was the preferred epitope for the immune recognition of sera specific for Cz and its C-T domain. Although to a minor extent, other anionic compounds bearing sulfate groups in different positions and number as well as different anionic charged groups including carboxylated or phosphorylated monosaccharides, disaccharides and oligosaccharides were recognized. In conclusion, we found that synthetic anionic sugar conjugates containing N-acetyl d-glucosamine-6-sulfate sodium salt (GlcNAc6S) competitively inhibit the binding of affinity purified rabbit anti-C-T IgG to the C-T extension of Cz. Extending these findings to the context of natural infection, immune assays performed with Chagas disease serum confirmed that the structure of synthetic GlcNAc6S mimics the N-glycan-linked sulfated epitope displayed in the C-T domain of Cz.