RESUMEN
In the present work we have investigated the serum antibody spectrum to parasite antigens involved in human T. cruzi infection. Analysis was performed by conventional serology (IHA, IFAT and ELISA), complement-mediated lysis, anti-gal antibody assay and reactivity against recombinant and synthetic peptides and metacyclic antigens by immunowestern-blotting. All the sera showed a significant reactivity in IHA, IFAT and ELISA. We found that 84.2% of the sera showed lytic activity and thirty serum samples (78.9%) which showed a lytic activity higher than 50%, also showed anti-gal antibodies at serum dilutions higher than 1:1,600. Ninety-four percent of sera reacted with one or more of the recombinant DNA clones and 97.3% reacted with one or more of the synthetic peptides. A pool of serum samples with a lytic activity higher than 75% were able to produce 60% to 78% inhibition of cell invasion. Thirty-six of the serum samples (94.7%) were able to react by immunowestern blotting with a T. cruzi metacyclic antigen with molecular size of 70 kDa. The results obtained give preliminary information about the humoral immune response and the possible role of antibodies in protection against T. cruzi infection of chronic patients from the highlands of Chile.
Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Antígenos de Protozoos/inmunología , Enfermedad de Chagas/inmunología , Trypanosoma cruzi/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Animales , Formación de Anticuerpos , Enfermedad de Chagas/sangre , Chile , Ensayo de Inmunoadsorción Enzimática , Femenino , Técnica del Anticuerpo Fluorescente Indirecta , Pruebas de Hemaglutinación , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Twenty-one Trypanosoma cruzi stocks isolated from Triatoma infestans and humans of the Chilean Andean highlands were studied at the genotypic level by schizodeme and molecular karyotype analyses, which allowed a clear distinction of the parasites from those hosts. A phenotypical characterization was performed by proteolytic activity after electrophoretic fractionation without discrimination among the stocks. Metacyclic trypomastigotes obtained in vitro proved to be infective to Swiss mice and the study of immune response and biological behavior was assessed. Of a total of 21 T. cruzi stocks, only 11 proved to be infective in mice due to difficulties in obtaining metacyclic trypomastigotes with the parasite populations isolated from humans. Western blot analysis revealed a complex immune response even in the first days postinfection with each T. cruzi strain studied. Antigenic recognition by each immune serum is characteristic, although several major and common antigens were detected. Lytic antibodies were studied by the in vitro complement-mediated lytic assay using purified metacyclic trypomastigotes as target cells. All the T. cruzi isolates tested induced lytic antibodies in this experimental model. Parasitemias were moderate and characteristic for each T. cruzi strain. Results are compared with metacyclic forms of the infective and pathogenic Tulahuen strain.