Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting)
Autor(a) principal: | |
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Data de Publicação: | 1990 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Revista do Instituto de Medicina Tropical de São Paulo |
Texto Completo: | https://www.revistas.usp.br/rimtsp/article/view/28765 |
Resumo: | A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained. |
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Revista do Instituto de Medicina Tropical de São Paulo |
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Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) Identificação simultânea de antígenos internos e de superfície de Trypanosoma cruzi reativos para diferentes classes de imunoglobulinas (radio-immunoblotting) Trypanosoma cruziChagas' diseaseSurface antigensWestern-blotImmunoblottingIgG classes A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained. Classes e subclasses de anticorpos apresentam diferentes funções, influenciando a resposta imune humoral de um hospedeiro, frente a um agente infeccioso. Na maioria dos sistemas, o alvo principal é representado pelos antígenos de membrana do parasita. Entretanto, a identificação de antígenos de superfície de parasitas, reativos para classe (e subclasse) de imunoglobulinas que não se ligam a proteína-A implica em imunoprecipitações sucessivas, que levam a perda de antígenos e/ou reações inespecíficas. Visando esse estudo, foi desenvolvida uma técnica denominada "radio-immunoblotting", através da qual a reatividade de imunoglobulinas de diferentes classes para antígenos de membrana (e/ou internos) foi analisada simultaneamente. O método constitui na marcação prévia da superfície dos parasitas por radioiodação, fracionamento dos polipeptídeos por SDS/PA-GE, transferência das frações para nitrocelulose, reação com soros e conjugados anti-Igs - peroxidase e autoradiografia dos mesmos, a análise é feita comparando-se os antígenos comuns evidenciados na autoradiografia e nas tiras de nitrocelulose coradas com o substrato da peroxidase. Essa técnica foi utilizada para analisar antígenos de superfície de formas tripomastigotas de T. cruzi reativas para IgG, IgM e IgA provenientes de soros de pacientes com doença de Chagas na fase aguda. Obtiveram-se distintos padrões de reatividade para as diferentes classes de anticorpos provenientes de um mesmo soro humano. Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo1990-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/rimtsp/article/view/28765Revista do Instituto de Medicina Tropical de São Paulo; Vol. 32 No. 5 (1990); 379-383 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 32 Núm. 5 (1990); 379-383 Revista do Instituto de Medicina Tropical de São Paulo; v. 32 n. 5 (1990); 379-383 1678-99460036-4665reponame:Revista do Instituto de Medicina Tropical de São Pauloinstname:Instituto de Medicina Tropical (IMT)instacron:IMTenghttps://www.revistas.usp.br/rimtsp/article/view/28765/30618Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Pauloinfo:eu-repo/semantics/openAccessStolf, Anna Maria SimonsenUmezawa, Eufrosina SetsuZingales, Bianca2012-07-02T01:28:43Zoai:revistas.usp.br:article/28765Revistahttp://www.revistas.usp.br/rimtsp/indexPUBhttps://www.revistas.usp.br/rimtsp/oai||revimtsp@usp.br1678-99460036-4665opendoar:2022-12-13T16:50:31.419912Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT)true |
dc.title.none.fl_str_mv |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) Identificação simultânea de antígenos internos e de superfície de Trypanosoma cruzi reativos para diferentes classes de imunoglobulinas (radio-immunoblotting) |
title |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) |
spellingShingle |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) Stolf, Anna Maria Simonsen Trypanosoma cruzi Chagas' disease Surface antigens Western-blot Immunoblotting IgG classes |
title_short |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) |
title_full |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) |
title_fullStr |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) |
title_full_unstemmed |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) |
title_sort |
Simultaneous identification of Trypanosoma cruzi surface and internal antigens reactive to different immunoglobulin classes (radio-immunoblotting) |
author |
Stolf, Anna Maria Simonsen |
author_facet |
Stolf, Anna Maria Simonsen Umezawa, Eufrosina Setsu Zingales, Bianca |
author_role |
author |
author2 |
Umezawa, Eufrosina Setsu Zingales, Bianca |
author2_role |
author author |
dc.contributor.author.fl_str_mv |
Stolf, Anna Maria Simonsen Umezawa, Eufrosina Setsu Zingales, Bianca |
dc.subject.por.fl_str_mv |
Trypanosoma cruzi Chagas' disease Surface antigens Western-blot Immunoblotting IgG classes |
topic |
Trypanosoma cruzi Chagas' disease Surface antigens Western-blot Immunoblotting IgG classes |
description |
A radioactive Western-blotting technique was developed by which the reactivity of Immunoglobulins (Igs) from different classes to both membrane radiolabelled and internal parasite antigens is simultaneously identified. The method includes radioiodination of parasites, polypeptide fractionation by SDS-PAGE, Western-blot transfer and autoradiography of the immunoblots developed with anti-Igs conjugates labelled with enzymes. The analysis is then performed by the comparison of common bands on the autoradiograms and the respective substrate stained nitrocellulose blots. This technique was used to analyse T. cruzi trypomastigote surface labelled antigens reactive to IgM, IgA and IgG specific antibodies. A different pattern of reactivity with acute Chagas' disease patients sera was thus obtained. |
publishDate |
1990 |
dc.date.none.fl_str_mv |
1990-10-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/28765 |
url |
https://www.revistas.usp.br/rimtsp/article/view/28765 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
https://www.revistas.usp.br/rimtsp/article/view/28765/30618 |
dc.rights.driver.fl_str_mv |
Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Copyright (c) 2018 Revista do Instituto de Medicina Tropical de São Paulo |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo |
publisher.none.fl_str_mv |
Universidade de São Paulo. Instituto de Medicina Tropical de São Paulo |
dc.source.none.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo; Vol. 32 No. 5 (1990); 379-383 Revista do Instituto de Medicina Tropical de São Paulo; Vol. 32 Núm. 5 (1990); 379-383 Revista do Instituto de Medicina Tropical de São Paulo; v. 32 n. 5 (1990); 379-383 1678-9946 0036-4665 reponame:Revista do Instituto de Medicina Tropical de São Paulo instname:Instituto de Medicina Tropical (IMT) instacron:IMT |
instname_str |
Instituto de Medicina Tropical (IMT) |
instacron_str |
IMT |
institution |
IMT |
reponame_str |
Revista do Instituto de Medicina Tropical de São Paulo |
collection |
Revista do Instituto de Medicina Tropical de São Paulo |
repository.name.fl_str_mv |
Revista do Instituto de Medicina Tropical de São Paulo - Instituto de Medicina Tropical (IMT) |
repository.mail.fl_str_mv |
||revimtsp@usp.br |
_version_ |
1798951638793191424 |