title: Immunoblotting analyses using two-dimensional gel electrophoresis of Trypanosoma cruzi excreted-secreted antigens.
abstract: Trypanosoma cruzi trypomastigotes excrete-secrete a complex mixture of antigenic molecules. This antigenic mixture denominated trypomastigote excreted-secreted antigens contains a 150-160 kDa band that shows excellent performance in Chagas' disease diagnosis by immunoblotting. The present study partially characterized by two-dimensional gel electrophoresis the immunoreactivity against the 150-160 kDa protein using sera samples from chagasic patients in different phases of the disease. Trypomastigote excreted-secreted antigen preparations were subjected to high-resolution two-dimensional (2D) gel electrophoresis followed by immunoblotting with sera from chagasic and non-chagasic patients. The 150-160 kDa protein presented four isoforms with isoelectric focusing ranging from 6.2 to 6.7. The four isoforms were recognized by IgM from acute phase and IgG from chronic phase sera of chagasic patients. The 150-160 kDa isoform with IF of approximately 6.4 became the immunodominant spot with the progression of the disease. No cross-reactivity was observed with non-chagasic or patients infected with Leishmania sp. In this study we provide basic knowledge that supports the validation of trypomastigote excreted-secreted antigens for serological diagnosis of Chagas' disease.
authors.affiliation: Laboratório de Imunologia do Instituto de Ciências Biomédicas da Universidade Federal de Uberlândia, Uberlândia, MG, Brasil.
authors.forename: Adriano Gomes
authors.initials: AG
authors.lastname: Silva
authors.forename: Elisangela Paula
authors.initials: EP
authors.lastname: Silveira-Lacerda
authors.forename: Jair Pereira
authors.initials: JP
authors.lastname: Cunha-Júnior
authors.forename: Maria Aparecida
authors.initials: MA
authors.lastname: de Souza
authors.forename: Silvio
authors.initials: S
authors.lastname: Favoreto Junior
mesh_terms: D000208:Acute Disease; D000818:Animals; D000953:Antigens, Protozoan / Q000097:blood*; D016022:Case-Control Studies; D014355:Chagas Disease / Q000175:diagnosis*; D002908:Chronic Disease; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D014349:Trypanosoma cruzi / Q000276:immunology*
chemical_list: D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D007075:Immunoglobulin M