Immunoblotting analyses using two-dimensional gel electrophoresis of Trypanosoma cruzi excreted-secreted antigens Análise de antígenos excretados-secretados de Trypanosoma cruzi por immunoblotting em gel bidimensional

• Published in: Revista da Sociedade Brasileira de Medicina Tropical
• Main Authors: Adriano Gomes Silva, Elisangela Paula Silveira-Lacerda, Jair Pereira Cunha-Júnior, Maria Aparecida de Souza, Silvio Favoreto Junior
• Format: Article in Journal/Newspaper
• Language: English
• Published: Sociedade Brasileira de Medicina Tropical (SBMT) 2004
• Subjects: Trypanosoma cruzi; Antígenos excretados; Eletroforese em duas dimensões; Immunoblotting; Excreted-secreted antigens; Two-dimensional gel electrophoresis; Arctic medicine. Tropical medicine; RC955-962; Arctic; Alta
• Online Access: https://doi.org/10.1590/S0037-86822004000600005; https://doaj.org/article/c78beecf3661406bb420f624527c58b0

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 150-160kDa 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 150-160kDa 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 150-160kDa 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. Formas tripomastigotas de Trypanosoma cruzi excretam/secretam uma complexa mistura de moléculas antigênicas. Essa mistura é chamada trypomastigote excreted-secreted antigens e contém uma banda de massa molecular em torno de 150-160kDa que possui excelente performance para diagnóstico de doença de Chagas em immunoblotting. No presente estudo foi caracterizado parcialmente, por gel bidimensional, a proteína de 150-160kDa pela análise da reatividade de anticorpos de pacientes chagásicos nas diversas fases da doença. Proteínas do trypomastigote excreted-secreted antigens foram separadas por eletroforese de alta resolução em duas dimensões (2D) e submetidas a immunoblotting com soros de pacientes chagásicos e não chagásicos. A proteína de 150-160kDa foi ...