Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans

Detalhes bibliográficos
Autor(a) principal: Monte, Leonardo Garcia
Data de Publicação: 2009
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFPel - Guaiaca
Texto Completo: http://guaiaca.ufpel.edu.br/handle/123456789/1263
Resumo: Leptospirosis is a zoonotic disease caused by pathogenic bacteria belonging to the Leptospira genus. Several mammals may carry the agent, and rats are the most important source of human infection in urban settings. The wide spectrum of clinical manifestations varies from mild cases, with fever and headaches, to severe presentations, with liver and kidney failure, which may lead to death. As a result of the various degrees of severity, leptospirosis is frequently mistaken, in its acute stage, with other tropical diseases such as influenza and dengue. The microscopic agglutination test (MAT) is considered the gold standard when diagnosing leptospirosis; however, the test presents limitations regarding sensitivity in the acute phase of the disease. Recently, surface proteins LigA and LigB have been identified to be related with leptospiral virulence. These proteins have been characterized as adhesins, with a proteic structure similar to Escherichia coli intimin and Yersinia pseudotuberculosis invasin, which are important virulence factors in these organisms. The goal of this study was to produce and characterize monoclonal antibodies (MAbs) against a truncated fragment of approximately 54 kDa, named rLigBrep, that comprise a identical portion of LigA and LigB (domains 2-7). The 5 MAbs obtained were of the IgG1 (2) and IgG2b (3) isotypes and their affinity constants for rLigBrep varied from 7 x 107 M-1 to 4 x 108 M-1. The MAbs were able to react with the native antigen in L. interrogans serovar Copenhageni strain Fiocruz L1-130 by indirect immunofluorescence, immunoblotting, whole-cell ELISA and immunoelectron microscopy. These results allow concluding that these MAbs are important tools for studies aiming understanding the role of Lig proteins in Leptospira pathogenesis and in the development of tests for diagnosis of leptospirosis.
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spelling 2014-08-20T13:32:53Z2010-03-182014-08-20T13:32:53Z2009-05-11MONTE, Leonardo Garcia. Production and characterization of monoclonal antibodies against the identical region of LigA and LigB proteins from Leptospira interrogans. 2009. 53 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2009.http://guaiaca.ufpel.edu.br/handle/123456789/1263Leptospirosis is a zoonotic disease caused by pathogenic bacteria belonging to the Leptospira genus. Several mammals may carry the agent, and rats are the most important source of human infection in urban settings. The wide spectrum of clinical manifestations varies from mild cases, with fever and headaches, to severe presentations, with liver and kidney failure, which may lead to death. As a result of the various degrees of severity, leptospirosis is frequently mistaken, in its acute stage, with other tropical diseases such as influenza and dengue. The microscopic agglutination test (MAT) is considered the gold standard when diagnosing leptospirosis; however, the test presents limitations regarding sensitivity in the acute phase of the disease. Recently, surface proteins LigA and LigB have been identified to be related with leptospiral virulence. These proteins have been characterized as adhesins, with a proteic structure similar to Escherichia coli intimin and Yersinia pseudotuberculosis invasin, which are important virulence factors in these organisms. The goal of this study was to produce and characterize monoclonal antibodies (MAbs) against a truncated fragment of approximately 54 kDa, named rLigBrep, that comprise a identical portion of LigA and LigB (domains 2-7). The 5 MAbs obtained were of the IgG1 (2) and IgG2b (3) isotypes and their affinity constants for rLigBrep varied from 7 x 107 M-1 to 4 x 108 M-1. The MAbs were able to react with the native antigen in L. interrogans serovar Copenhageni strain Fiocruz L1-130 by indirect immunofluorescence, immunoblotting, whole-cell ELISA and immunoelectron microscopy. These results allow concluding that these MAbs are important tools for studies aiming understanding the role of Lig proteins in Leptospira pathogenesis and in the development of tests for diagnosis of leptospirosis.A leptospirose é uma zoonose causada por bactérias patogênicas pertencentes ao gênero Leptospira. Diversos mamíferos podem albergar o agente, sendo o rato a principal fonte de infecção para humanos no ambiente. As manifestações clínicas da doença variam desde os sintomas leves, como febre e dores de cabeça, até os mais graves com insuficiência renal e hepática que podem levar o indivíduo à morte. Esse amplo espectro de sintomas faz com que a leptospirose seja freqüentemente confundida em sua fase aguda com outras doenças febris, como gripe e dengue. O teste de aglutinação microscópica (MAT) é considerado o padrão ouro para o diagnóstico laboratorial da leptospirose, entretanto, o teste apresenta limitações relacionadas com a sensibilidade na fase aguda da doença. Recentemente, as proteínas de superfície LigA e LigB foram identificadas e relacionadas com a virulência de leptospiras patogênicas. Estas proteínas foram caracterizadas como adesinas, possuindo estrutura protéica semelhante a intimina de Escherichia coli e invasina de Yersinia pseudotuberculosis, que são importantes fatores de virulência nestes microrganismos. O objetivo desse estudo foi produzir e caracterizar anticorpos monoclonais (MAbs) contra um fragmento truncado de aproximadamente 54 kDa, que codifica a região idêntica das proteínas LigA e LigB (domínios 2-7), denominado rLigBrep. Foram obtidos 5 MAbs dos isotipos IgG1 (2) e IgG2b (3), com afinidades por rLigBrep que variaram entre 7 x 107 M-1 e 4 x 108 M-1. Foi comprovado através de imunofluorescência indireta, immunoblotting, ELISA whole-cell e microscopia imunoeletrônica que os MAbs foram capazes de detectar o antígeno nativo presente em L. interrogans sorovar Copenhageni cepa Fiocruz L1-130. Estes resultados permitem concluir que os MAbs produzidos são importantes ferramentas para serem utilizados em estudos que visam entender o papel das proteínas Lig na patogênese das leptospiras e em testes diagnósticos para leptospirose.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaBiotechnologyLeptospirosisMonoclonal antibodiesDiagnosisBiotecnologiaLeptospiroseAnticorpos monoclonaisDiagnósticoCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIAProdução e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogansProduction and characterization of monoclonal antibodies against the identical region of LigA and LigB proteins from Leptospira interrogansinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://lattes.cnpq.br/3215575195506213http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783342J3&dataRevisao=nullLeite, Fabio Pereira Leivashttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763076J5Aleixo, José Antônio GuimarãesMonte, Leonardo Garciainfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALdissertacao_leonardo_monte.pdfapplication/pdf1685536http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1263/1/dissertacao_leonardo_monte.pdf2128191ebf9a0b395668fcbb7af16da9MD51open accessTEXTdissertacao_leonardo_monte.pdf.txtdissertacao_leonardo_monte.pdf.txtExtracted Texttext/plain68859http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1263/2/dissertacao_leonardo_monte.pdf.txt193df7e757a11665e5d9fed40d306345MD52open accessTHUMBNAILdissertacao_leonardo_monte.pdf.jpgdissertacao_leonardo_monte.pdf.jpgGenerated Thumbnailimage/jpeg1790http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1263/3/dissertacao_leonardo_monte.pdf.jpg001fbb6a95f755a9832d311649fdcc9cMD53open access123456789/12632019-08-23 10:38:55.699open accessoai:guaiaca.ufpel.edu.br:123456789/1263Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T13:38:55Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false
dc.title.por.fl_str_mv Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
dc.title.alternative.eng.fl_str_mv Production and characterization of monoclonal antibodies against the identical region of LigA and LigB proteins from Leptospira interrogans
title Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
spellingShingle Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
Monte, Leonardo Garcia
Biotechnology
Leptospirosis
Monoclonal antibodies
Diagnosis
Biotecnologia
Leptospirose
Anticorpos monoclonais
Diagnóstico
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
title_short Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
title_full Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
title_fullStr Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
title_full_unstemmed Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
title_sort Produção e caracterização de anticorpos monoclonais contra a região idêntica das proteínas LigA e LigB de Leptospira interrogans
author Monte, Leonardo Garcia
author_facet Monte, Leonardo Garcia
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/3215575195506213
dc.contributor.advisorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783342J3&dataRevisao=null
dc.contributor.referees1.pt_BR.fl_str_mv Leite, Fabio Pereira Leivas
dc.contributor.referees1ID.por.fl_str_mv
dc.contributor.referees1Lattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763076J5
dc.contributor.advisor1.fl_str_mv Aleixo, José Antônio Guimarães
dc.contributor.author.fl_str_mv Monte, Leonardo Garcia
contributor_str_mv Aleixo, José Antônio Guimarães
dc.subject.eng.fl_str_mv Biotechnology
Leptospirosis
Monoclonal antibodies
Diagnosis
topic Biotechnology
Leptospirosis
Monoclonal antibodies
Diagnosis
Biotecnologia
Leptospirose
Anticorpos monoclonais
Diagnóstico
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
dc.subject.por.fl_str_mv Biotecnologia
Leptospirose
Anticorpos monoclonais
Diagnóstico
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA
description Leptospirosis is a zoonotic disease caused by pathogenic bacteria belonging to the Leptospira genus. Several mammals may carry the agent, and rats are the most important source of human infection in urban settings. The wide spectrum of clinical manifestations varies from mild cases, with fever and headaches, to severe presentations, with liver and kidney failure, which may lead to death. As a result of the various degrees of severity, leptospirosis is frequently mistaken, in its acute stage, with other tropical diseases such as influenza and dengue. The microscopic agglutination test (MAT) is considered the gold standard when diagnosing leptospirosis; however, the test presents limitations regarding sensitivity in the acute phase of the disease. Recently, surface proteins LigA and LigB have been identified to be related with leptospiral virulence. These proteins have been characterized as adhesins, with a proteic structure similar to Escherichia coli intimin and Yersinia pseudotuberculosis invasin, which are important virulence factors in these organisms. The goal of this study was to produce and characterize monoclonal antibodies (MAbs) against a truncated fragment of approximately 54 kDa, named rLigBrep, that comprise a identical portion of LigA and LigB (domains 2-7). The 5 MAbs obtained were of the IgG1 (2) and IgG2b (3) isotypes and their affinity constants for rLigBrep varied from 7 x 107 M-1 to 4 x 108 M-1. The MAbs were able to react with the native antigen in L. interrogans serovar Copenhageni strain Fiocruz L1-130 by indirect immunofluorescence, immunoblotting, whole-cell ELISA and immunoelectron microscopy. These results allow concluding that these MAbs are important tools for studies aiming understanding the role of Lig proteins in Leptospira pathogenesis and in the development of tests for diagnosis of leptospirosis.
publishDate 2009
dc.date.issued.fl_str_mv 2009-05-11
dc.date.available.fl_str_mv 2010-03-18
2014-08-20T13:32:53Z
dc.date.accessioned.fl_str_mv 2014-08-20T13:32:53Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
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dc.identifier.citation.fl_str_mv MONTE, Leonardo Garcia. Production and characterization of monoclonal antibodies against the identical region of LigA and LigB proteins from Leptospira interrogans. 2009. 53 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2009.
dc.identifier.uri.fl_str_mv http://guaiaca.ufpel.edu.br/handle/123456789/1263
identifier_str_mv MONTE, Leonardo Garcia. Production and characterization of monoclonal antibodies against the identical region of LigA and LigB proteins from Leptospira interrogans. 2009. 53 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2009.
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publisher.none.fl_str_mv Universidade Federal de Pelotas
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