Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFPel - Guaiaca |
Texto Completo: | http://guaiaca.ufpel.edu.br/handle/123456789/1285 |
Resumo: | Leptospirosis is an infectious disease that affects humans, wild and domestic animals worldwide. Pathogenic spirochetes from the Leptospira genus are the causative agents of this zoonosis. The several Leptospira species have noted antigenic diversity, even within the same species. This is the main reason current bacterin vaccines have limitations, such as adverse effects and short term immunity, restricting their use in human populations. The need for effective leptospirosis vaccines promoted studies on characterization of new vaccine candidates. The 32 kDa outer membrane lipoprotein, LipL32, is the most abundant protein in the whole leptospira proteome, it is conserved in all pathogenic serovars and absent in saprophytes. This protein is immunogenic and able to bind to mammalian extracellular matrix. However, LipL32 subunit vaccines did not protect animals against challenge. In an attempt to solve this, we use LipL32 fused and coadministered with B subunit of the Escherichia coli heat-labile enterotoxin (LTB) to enhance the immune response. LTB is a non-toxic molecule with immunoestimulatory and immunomodulatory properties. The recombinant proteins rLTB, rLipL32 and rLTB::LipL32 were expressed in E. coli, purified and characterized. Female hamsters were distributed in groups as follows: rLTB; rLTB+rLipL32; rLTB::LipL32, homologous bacterin; PBS. The serum from each animal was collected for humoral immune response determination by ELISA. The animals were challenged with 5×LD50 dose of Leptospira interrogans strain Fiocruz L1-130. Both treatments induced high titers of anti-rLipL32 antibodies. The rLTB+rLipL32 and rLTB::LipL32 treatments afforded significant protective response upon challenge, when compared to control groups (p<0.05). No prior study with leptospirosis had used LTB as the adjuvant, or fused antigens in an attempt to control this disease. Furthermore, this is the first report of a protective subunit vaccine using rLipL32 as the antigen, and an important contribution towards the development of improved leptospirosis vaccines. |
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2014-08-20T13:32:57Z2011-05-162014-08-20T13:32:57Z2011-02-28GRASSMANN, André Alex. Protection against leptospirosis induced by LipL32 co-administered or fused to LTB. 2011. 51 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2011.http://guaiaca.ufpel.edu.br/handle/123456789/1285Leptospirosis is an infectious disease that affects humans, wild and domestic animals worldwide. Pathogenic spirochetes from the Leptospira genus are the causative agents of this zoonosis. The several Leptospira species have noted antigenic diversity, even within the same species. This is the main reason current bacterin vaccines have limitations, such as adverse effects and short term immunity, restricting their use in human populations. The need for effective leptospirosis vaccines promoted studies on characterization of new vaccine candidates. The 32 kDa outer membrane lipoprotein, LipL32, is the most abundant protein in the whole leptospira proteome, it is conserved in all pathogenic serovars and absent in saprophytes. This protein is immunogenic and able to bind to mammalian extracellular matrix. However, LipL32 subunit vaccines did not protect animals against challenge. In an attempt to solve this, we use LipL32 fused and coadministered with B subunit of the Escherichia coli heat-labile enterotoxin (LTB) to enhance the immune response. LTB is a non-toxic molecule with immunoestimulatory and immunomodulatory properties. The recombinant proteins rLTB, rLipL32 and rLTB::LipL32 were expressed in E. coli, purified and characterized. Female hamsters were distributed in groups as follows: rLTB; rLTB+rLipL32; rLTB::LipL32, homologous bacterin; PBS. The serum from each animal was collected for humoral immune response determination by ELISA. The animals were challenged with 5×LD50 dose of Leptospira interrogans strain Fiocruz L1-130. Both treatments induced high titers of anti-rLipL32 antibodies. The rLTB+rLipL32 and rLTB::LipL32 treatments afforded significant protective response upon challenge, when compared to control groups (p<0.05). No prior study with leptospirosis had used LTB as the adjuvant, or fused antigens in an attempt to control this disease. Furthermore, this is the first report of a protective subunit vaccine using rLipL32 as the antigen, and an important contribution towards the development of improved leptospirosis vaccines.A leptospirose é uma doença infecciosa que afeta humanos e animais silvestres e domésticos em todo mundo. As espiroquetas patogênicas do gênero Leptospira são os agentes causadores desta zoonose. As diversas espécies de leptospiras possuem notada diversidade antigênica, inclusive em uma mesma espécie. Esta característica resulta em limitação das atuais vacinas bacterinas que não induzem proteção cruzada entre os diferentes sorovares. Além disso, estas vacinas geram efeitos adversos e imunidade de curta duração, restringindo seu uso em populações humanas. A necessidade de novas vacinas eficazes contra a leptospirose estimulou estudos para caracterizar novos antígenos vacinais. A lipoproteína de membrana externa de 32 kDa, LipL32 é a proteína mais abundante no proteoma total da leptospira, conservada entre todos os sorovares patogênicos e ausente nas leptospiras saprófitas. Esta proteína é imunogênica e possui habilidade de ligar-se à matriz extracelular de mamíferos. Porém, animais inoculados com vacinas de subunidade utilizando LipL32 não sobrevivem ao desafio. Em função disso, utilizamos LipL32 fusionada e co-administrada com a subunidade B da enterotoxina termolábil de Escherichia coli (LTB) para melhorar a resposta imune. LTB é uma molécula atóxica com reconhecida atividade imunoestimuladora e imunomoduladora. As proteínas recombinantes rLTB, rLipL32 e rLTB::LipL32 foram produzidas em E. coli, purificadas e caracterizadas. Hamsters fêmeas foram distribuídas em grupos e inoculadas com duas doses, da seguinte forma: rLTB; rLTB+rLipL32; rLTB::LipL32, bacterina homóloga e PBS. Soro foi coletado individualmente para determinação da resposta imune humoral por ELISA. Os animais foram desafiados com uma dose de 5×DL50 de Leptospira interrogans cepa Fiocruz L1-130. Os tratamentos induziram altos títulos de anticorpos anti-rLipL32. Os tratamentos rLTB+rLipL32 e rLTB::LipL32 induziram resposta protetora significativa frente ao desafio quando comparados com os grupos controle (p<0,05). Nenhum estudo anterior usou LTB como adjuvante para uma vacina contra leptospirose, tampouco antígenos fusionados com o intuito de controlar esta doença. Além disso, este é o primeiro relato de indução de imunidade protetora utilizando rLipL32 como vacina de subunidade, uma importante contribuição para o desenvolvimento de vacinas mais eficazes contra leptospirose.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaLeptospirosisRecombinant vaccineLeptospiraLeptospiroseVacina recombinanteLTBLipL32CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULARProteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTBProtection against leptospirosis induced by LipL32 co-administered or fused to LTBinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesishttp://lattes.cnpq.br/3880433527026304http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9Dellagostin, Odir AntônioGrassmann, André Alexinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALdissertacao_andre_grassmann.pdfapplication/pdf1497268http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1285/1/dissertacao_andre_grassmann.pdf73f55a329ef2c9be70227b63b45785a2MD51open accessTEXTdissertacao_andre_grassmann.pdf.txtdissertacao_andre_grassmann.pdf.txtExtracted Texttext/plain90702http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1285/2/dissertacao_andre_grassmann.pdf.txtdac7b6bf0e361d0b538c9f30eae8f670MD52open accessTHUMBNAILdissertacao_andre_grassmann.pdf.jpgdissertacao_andre_grassmann.pdf.jpgGenerated Thumbnailimage/jpeg1860http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1285/3/dissertacao_andre_grassmann.pdf.jpg663448a1415c5c7e9666debd075c72e3MD53open access123456789/12852019-08-23 09:27:43.367open accessoai:guaiaca.ufpel.edu.br:123456789/1285Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T12:27:43Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false |
dc.title.por.fl_str_mv |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
dc.title.alternative.eng.fl_str_mv |
Protection against leptospirosis induced by LipL32 co-administered or fused to LTB |
title |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
spellingShingle |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB Grassmann, André Alex Leptospirosis Recombinant vaccine Leptospira Leptospirose Vacina recombinante LTB LipL32 CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR |
title_short |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
title_full |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
title_fullStr |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
title_full_unstemmed |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
title_sort |
Proteção contra leptospirose induzida por LipL32 coadministrada ou fusionada à LTB |
author |
Grassmann, André Alex |
author_facet |
Grassmann, André Alex |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://lattes.cnpq.br/3880433527026304 |
dc.contributor.advisorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9 |
dc.contributor.advisor1.fl_str_mv |
Dellagostin, Odir Antônio |
dc.contributor.author.fl_str_mv |
Grassmann, André Alex |
contributor_str_mv |
Dellagostin, Odir Antônio |
dc.subject.eng.fl_str_mv |
Leptospirosis Recombinant vaccine |
topic |
Leptospirosis Recombinant vaccine Leptospira Leptospirose Vacina recombinante LTB LipL32 CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR |
dc.subject.por.fl_str_mv |
Leptospira Leptospirose Vacina recombinante LTB LipL32 |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::BIOQUIMICA::BIOLOGIA MOLECULAR |
description |
Leptospirosis is an infectious disease that affects humans, wild and domestic animals worldwide. Pathogenic spirochetes from the Leptospira genus are the causative agents of this zoonosis. The several Leptospira species have noted antigenic diversity, even within the same species. This is the main reason current bacterin vaccines have limitations, such as adverse effects and short term immunity, restricting their use in human populations. The need for effective leptospirosis vaccines promoted studies on characterization of new vaccine candidates. The 32 kDa outer membrane lipoprotein, LipL32, is the most abundant protein in the whole leptospira proteome, it is conserved in all pathogenic serovars and absent in saprophytes. This protein is immunogenic and able to bind to mammalian extracellular matrix. However, LipL32 subunit vaccines did not protect animals against challenge. In an attempt to solve this, we use LipL32 fused and coadministered with B subunit of the Escherichia coli heat-labile enterotoxin (LTB) to enhance the immune response. LTB is a non-toxic molecule with immunoestimulatory and immunomodulatory properties. The recombinant proteins rLTB, rLipL32 and rLTB::LipL32 were expressed in E. coli, purified and characterized. Female hamsters were distributed in groups as follows: rLTB; rLTB+rLipL32; rLTB::LipL32, homologous bacterin; PBS. The serum from each animal was collected for humoral immune response determination by ELISA. The animals were challenged with 5×LD50 dose of Leptospira interrogans strain Fiocruz L1-130. Both treatments induced high titers of anti-rLipL32 antibodies. The rLTB+rLipL32 and rLTB::LipL32 treatments afforded significant protective response upon challenge, when compared to control groups (p<0.05). No prior study with leptospirosis had used LTB as the adjuvant, or fused antigens in an attempt to control this disease. Furthermore, this is the first report of a protective subunit vaccine using rLipL32 as the antigen, and an important contribution towards the development of improved leptospirosis vaccines. |
publishDate |
2011 |
dc.date.available.fl_str_mv |
2011-05-16 2014-08-20T13:32:57Z |
dc.date.issued.fl_str_mv |
2011-02-28 |
dc.date.accessioned.fl_str_mv |
2014-08-20T13:32:57Z |
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 |
GRASSMANN, André Alex. Protection against leptospirosis induced by LipL32 co-administered or fused to LTB. 2011. 51 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2011. |
dc.identifier.uri.fl_str_mv |
http://guaiaca.ufpel.edu.br/handle/123456789/1285 |
identifier_str_mv |
GRASSMANN, André Alex. Protection against leptospirosis induced by LipL32 co-administered or fused to LTB. 2011. 51 f. Dissertação (Mestrado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2011. |
url |
http://guaiaca.ufpel.edu.br/handle/123456789/1285 |
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por |
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por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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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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