Avaliação de vacinas recombinantes contra a leptospirose
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFPel - Guaiaca |
Texto Completo: | http://repositorio.ufpel.edu.br/handle/ri/1220 |
Resumo: | Leptospirosis is considered a global problem regarding both veterinary medicine and public health. In Brazil, the disease has greater impact in vulnerable populations, living in slums of great cities, where high morbidity and mortality occur. This situation requires the establishment of new intervention policies in order to reduce cases. Available vaccines to prevent the disease in humans and animals are notoriously underachieving, producing short term, serovar specific protection and collateral effects. New strategies are being employed to develop novel vaccines against leptospirosis, such as the characterization of recombinant proteins, construction of DNA vaccines, and use of alternative adjuvants. Surface exposed proteins LipL32, LigB, and LigA are highly conserved and found only in pathogenic serovars, therefore these were selected to be used in this study. LigA and LipL32 genes were cloned into the pVAX eukaryote expression vector to be used as DNA vaccines. rLigBNI and rLipL32 were assessed as subunit recombinant antigens using propolis as a co-adjuvant. Hamsters were immunized and the response was assessed through qPCR, ELISA, and lethal challenge. DNA vaccine containing the LipL32 gene, and the subunit rLigBNI vaccine had the highest results in the immune-stimulation assays. Furthermore, these imunogens were able to significantly protect animals against lethal challenge, demonstrating their potential for leptospirosis vaccine development. |
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2014-08-20T13:32:48Z2013-11-252014-08-20T13:32:48Z2012-02-29FAGUNDES, Michel Quevedo. Avaliação de vacinas recombinantes contra a leptospirose. 2012. 62 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2012.http://repositorio.ufpel.edu.br/handle/ri/1220Leptospirosis is considered a global problem regarding both veterinary medicine and public health. In Brazil, the disease has greater impact in vulnerable populations, living in slums of great cities, where high morbidity and mortality occur. This situation requires the establishment of new intervention policies in order to reduce cases. Available vaccines to prevent the disease in humans and animals are notoriously underachieving, producing short term, serovar specific protection and collateral effects. New strategies are being employed to develop novel vaccines against leptospirosis, such as the characterization of recombinant proteins, construction of DNA vaccines, and use of alternative adjuvants. Surface exposed proteins LipL32, LigB, and LigA are highly conserved and found only in pathogenic serovars, therefore these were selected to be used in this study. LigA and LipL32 genes were cloned into the pVAX eukaryote expression vector to be used as DNA vaccines. rLigBNI and rLipL32 were assessed as subunit recombinant antigens using propolis as a co-adjuvant. Hamsters were immunized and the response was assessed through qPCR, ELISA, and lethal challenge. DNA vaccine containing the LipL32 gene, and the subunit rLigBNI vaccine had the highest results in the immune-stimulation assays. Furthermore, these imunogens were able to significantly protect animals against lethal challenge, demonstrating their potential for leptospirosis vaccine development.A leptospirose é considerada um problema global para a saúde pública e veterinária. No Brasil, a enfermidade possui maior impacto em populações vulneráveis, as quais estão distribuídas nas favelas das grandes cidades, onde tradicionalmente ocorre elevada morbidade e mortalidade. Desta forma, torna-se necessário o estabelecimento de novas efetivas de intervenção para a redução dos casos. As vacinas disponíveis para a prevenção da enfermidade em humanos e animais são comprovadamente limitadas, já que induzem imunidade pouco duradoura e sorovar-específica, além de provocarem efeitos colaterais. Novas estratégias para o desenvolvimento de uma vacina contra a leptospirose têm sido empregadas, como a caracterização de proteínas recombinantes, a construção de vacinas de DNA e o teste de novos adjuvantes para modulação da resposta imune. Neste contexto, as proteínas de membrana externa LipL32, LigB e LigA foram selecionadas para este trabalho, pois são conservadas e encontradas somente entre sorovares patogênicos de Leptospira. Assim, os genes lipL32 e ligA foram clonados no vetor de expressão em eucariotos pVAX para a obtenção de vacinas de DNA, e as proteínas rLigBNI e rLipL32 foram testadas utilizando o própolis como co-adjuvante na forma de vacinas de subunidade. Após a produção das vacinas, hamsters foram vacinados e a resposta imune foi avaliada através de ELISA, qPCR e desafio. Dentre as preparações testadas, a resposta imune teve níveis mais elevados e significativos para a vacina de DNA contendo o gene lipL32 e para a vacina de subunidade de LigBNI. Além disso, estas vacinas foram capazes de proteger significativamente os animais contra o desafio homólogo, mostrando o potencial destas formulações para o desenvolvimento de uma vacina recombinante para a leptospirose.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaLeptospiroseProteínas recombinantesVacinas de DNAPrópolisLeptospirosisRecombinant vaccinesDNA vaccinesPropolisCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIAAvaliação de vacinas recombinantes contra a leptospiroseAvaliação de vacinas recombinantes contra a leptospiroseinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://lattes.cnpq.br/8739275051188774http://lattes.cnpq.br/5116470459238753Silva, Éverton Fagonde daFagundes, Michel Quevedoinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALdissertacao_michel_quevedo_fagundes.pdfapplication/pdf823551http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1220/1/dissertacao_michel_quevedo_fagundes.pdf36c28ca5bd55c0c0403915db568e103eMD51open accessTEXTdissertacao_michel_quevedo_fagundes.pdf.txtdissertacao_michel_quevedo_fagundes.pdf.txtExtracted Texttext/plain104911http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1220/2/dissertacao_michel_quevedo_fagundes.pdf.txt0911ee61e34e32e83422b4e66704e199MD52open accessTHUMBNAILdissertacao_michel_quevedo_fagundes.pdf.jpgdissertacao_michel_quevedo_fagundes.pdf.jpgGenerated Thumbnailimage/jpeg1924http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1220/3/dissertacao_michel_quevedo_fagundes.pdf.jpg1b94c22ae0b0e97a07b45fba73a3ec7dMD53open access123456789/12202019-08-23 10:01:53.226open accessoai:guaiaca.ufpel.edu.br:123456789/1220Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T13:01:53Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false |
dc.title.por.fl_str_mv |
Avaliação de vacinas recombinantes contra a leptospirose |
dc.title.alternative.eng.fl_str_mv |
Avaliação de vacinas recombinantes contra a leptospirose |
title |
Avaliação de vacinas recombinantes contra a leptospirose |
spellingShingle |
Avaliação de vacinas recombinantes contra a leptospirose Fagundes, Michel Quevedo Leptospirose Proteínas recombinantes Vacinas de DNA Própolis Leptospirosis Recombinant vaccines DNA vaccines Propolis CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
title_short |
Avaliação de vacinas recombinantes contra a leptospirose |
title_full |
Avaliação de vacinas recombinantes contra a leptospirose |
title_fullStr |
Avaliação de vacinas recombinantes contra a leptospirose |
title_full_unstemmed |
Avaliação de vacinas recombinantes contra a leptospirose |
title_sort |
Avaliação de vacinas recombinantes contra a leptospirose |
author |
Fagundes, Michel Quevedo |
author_facet |
Fagundes, Michel Quevedo |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/8739275051188774 |
dc.contributor.advisorLattes.por.fl_str_mv |
http://lattes.cnpq.br/5116470459238753 |
dc.contributor.advisor1.fl_str_mv |
Silva, Éverton Fagonde da |
dc.contributor.author.fl_str_mv |
Fagundes, Michel Quevedo |
contributor_str_mv |
Silva, Éverton Fagonde da |
dc.subject.por.fl_str_mv |
Leptospirose Proteínas recombinantes Vacinas de DNA Própolis |
topic |
Leptospirose Proteínas recombinantes Vacinas de DNA Própolis Leptospirosis Recombinant vaccines DNA vaccines Propolis CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
dc.subject.eng.fl_str_mv |
Leptospirosis Recombinant vaccines DNA vaccines Propolis |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
description |
Leptospirosis is considered a global problem regarding both veterinary medicine and public health. In Brazil, the disease has greater impact in vulnerable populations, living in slums of great cities, where high morbidity and mortality occur. This situation requires the establishment of new intervention policies in order to reduce cases. Available vaccines to prevent the disease in humans and animals are notoriously underachieving, producing short term, serovar specific protection and collateral effects. New strategies are being employed to develop novel vaccines against leptospirosis, such as the characterization of recombinant proteins, construction of DNA vaccines, and use of alternative adjuvants. Surface exposed proteins LipL32, LigB, and LigA are highly conserved and found only in pathogenic serovars, therefore these were selected to be used in this study. LigA and LipL32 genes were cloned into the pVAX eukaryote expression vector to be used as DNA vaccines. rLigBNI and rLipL32 were assessed as subunit recombinant antigens using propolis as a co-adjuvant. Hamsters were immunized and the response was assessed through qPCR, ELISA, and lethal challenge. DNA vaccine containing the LipL32 gene, and the subunit rLigBNI vaccine had the highest results in the immune-stimulation assays. Furthermore, these imunogens were able to significantly protect animals against lethal challenge, demonstrating their potential for leptospirosis vaccine development. |
publishDate |
2012 |
dc.date.issued.fl_str_mv |
2012-02-29 |
dc.date.available.fl_str_mv |
2013-11-25 2014-08-20T13:32:48Z |
dc.date.accessioned.fl_str_mv |
2014-08-20T13:32:48Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
FAGUNDES, Michel Quevedo. Avaliação de vacinas recombinantes contra a leptospirose. 2012. 62 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.ufpel.edu.br/handle/ri/1220 |
identifier_str_mv |
FAGUNDES, Michel Quevedo. Avaliação de vacinas recombinantes contra a leptospirose. 2012. 62 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2012. |
url |
http://repositorio.ufpel.edu.br/handle/ri/1220 |
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Universidade Federal de Pelotas |
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Programa de Pós-Graduação em Biotecnologia |
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UFPel |
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BR |
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Biotecnologia |
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Universidade Federal de Pelotas |
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