Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose

Detalhes bibliográficos
Autor(a) principal: Bacelo, Kátia Leston
Data de Publicação: 2013
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFPel - Guaiaca
Texto Completo: http://guaiaca.ufpel.edu.br/handle/123456789/1227
Resumo: A major challenge for the development of vaccines based on purified or recombinant protein subunits, and synthetic peptides resides in the fact that these are poorly immunogenic and mobilize insufficient immunoprotective response. Adjuvants are often used in association with these subunits in order to amplify and direct the immune response induced. Nowadays, there is a wide range of compounds that demonstrate adjuvant activity, however, few are approved for human use, and these often fail to induce appropriate immune response against a particular pathogen. Thus, there is a need to develop new adjuvants that are safe, effective and represent an alternative to currently available. In the present study, we used as a model antigen the non- identical portion of the Leptospira LigA protein (Leptospiral immunoglobulin-like protein A), an outer membrane protein of great interest as a mediator pathogenic mechanisms, used in serological diagnosis and as experimental vaccines. This antigen was formulated with various adjuvants, and the formulations tested for their immunoprotective potential in hamsters, challenged with a virulent strain of L. interrogans serovar Copenhageni. For this, the LigAni protein was produced in recombinant form (rLigAni) using Escherichia coli as the expression system and associated to xanthan polysaccharide, in its variants xanthan pruni strains 106 (X1) and 101 (X2), commercial xanthan and also to oligodinucleotídeo CpG (CpG ODN), carbon nanotubes (CNTs) and aluminum hydroxide (Alhydrogel). Formulations containing rLigAni associated with xanthan polysaccharide and CNTs induced significant IgG antibody titers, comparable to that induced when the protein was associated with Alhydrogel. Protection against lethal challenge was observed in 100%, 100%, 67% and 50% of the hamsters immunized with rLigAni-X1, rLigAni- CpG-X1, rLigAni-Alhydrogel and rLigAni-X2, respectively (Fisher test P < 0.05). The preparations containing rLigAni associated with CNTs, although induced an antibody response, failed to confer immunoprotection. Additionally, xanthan and CNTs adjuvants were not toxic to Chinese hamster ovary (CHO) cells, in vitro. The results of this study indicate xanthan as a new adjuvant for subunit vaccines against leptospirosis, presenting the ability to potentiate the immune response against the antigen, besides biocompatibility and the possibility of reduction of number of doses required for protection.
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spelling http://lattes.cnpq.br/3849631792846037http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9Dellagostin, Odir AntônioBacelo, Kátia Leston2014-08-20T13:32:49Z2014-01-272014-08-20T13:32:49Z2013-12-10BACELO, Kátia Leston. Adjuvants: impact on the effectiveness in subunit vaccine against leptospirosis. 2013. 111 f. Tese (Doutorado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2013.http://guaiaca.ufpel.edu.br/handle/123456789/1227A major challenge for the development of vaccines based on purified or recombinant protein subunits, and synthetic peptides resides in the fact that these are poorly immunogenic and mobilize insufficient immunoprotective response. Adjuvants are often used in association with these subunits in order to amplify and direct the immune response induced. Nowadays, there is a wide range of compounds that demonstrate adjuvant activity, however, few are approved for human use, and these often fail to induce appropriate immune response against a particular pathogen. Thus, there is a need to develop new adjuvants that are safe, effective and represent an alternative to currently available. In the present study, we used as a model antigen the non- identical portion of the Leptospira LigA protein (Leptospiral immunoglobulin-like protein A), an outer membrane protein of great interest as a mediator pathogenic mechanisms, used in serological diagnosis and as experimental vaccines. This antigen was formulated with various adjuvants, and the formulations tested for their immunoprotective potential in hamsters, challenged with a virulent strain of L. interrogans serovar Copenhageni. For this, the LigAni protein was produced in recombinant form (rLigAni) using Escherichia coli as the expression system and associated to xanthan polysaccharide, in its variants xanthan pruni strains 106 (X1) and 101 (X2), commercial xanthan and also to oligodinucleotídeo CpG (CpG ODN), carbon nanotubes (CNTs) and aluminum hydroxide (Alhydrogel). Formulations containing rLigAni associated with xanthan polysaccharide and CNTs induced significant IgG antibody titers, comparable to that induced when the protein was associated with Alhydrogel. Protection against lethal challenge was observed in 100%, 100%, 67% and 50% of the hamsters immunized with rLigAni-X1, rLigAni- CpG-X1, rLigAni-Alhydrogel and rLigAni-X2, respectively (Fisher test P < 0.05). The preparations containing rLigAni associated with CNTs, although induced an antibody response, failed to confer immunoprotection. Additionally, xanthan and CNTs adjuvants were not toxic to Chinese hamster ovary (CHO) cells, in vitro. The results of this study indicate xanthan as a new adjuvant for subunit vaccines against leptospirosis, presenting the ability to potentiate the immune response against the antigen, besides biocompatibility and the possibility of reduction of number of doses required for protection.O maior desafio para o desenvolvimento de vacinas baseadas em subunidades proteicas, recombinantes ou purificadas e peptídeos sintéticos, reside no fato destas serem pouco imunogênicas e mobilizarem uma resposta imunoprotetora insuficiente. Adjuvantes são utilizados associados a estas subunidades com o intuito de amplificar e direcionar a resposta imune induzida. Na atualidade, existe uma grande gama de compostos que demostram ação adjuvante, contudo, poucos são aprovados para uso humano, e estes muitas vezes falham em induzir resposta imune adequada contra determinado agente patogênico. Assim, existe a necessidade do desenvolvimento de novos adjuvantes que sejam seguros, efetivos e representem uma alternativa aos atualmente disponíveis. No presente estudo, utilizamos como antígeno modelo a porção não-idêntica da proteína LigA (Leptospiral immunoglobulin-like protein A) de Leptospira spp., uma proteína de membrana externa de grande interesse como mediadora de mecanismos de patogenicidade, utilizada em sorodiagnóstico e em vacinas experimentais. Esta proteína foi associada a diferentes adjuvantes, e as formulações testadas quanto ao seu potencial imunoprotetor em hamsters desafiados com cepa virulenta de Leptospira interrogans sorovar Copenhageni. Para isso, a proteína LigAni foi produzida em sua forma recombinante (rLigAni) utilizando Escherichia coli como sistema de expressão e associada ao polissacarídeo xantana, em suas variantes xantana pruni cepas 106 (X1) e 101 (X2), xantana comercial, e também ao oligodinucleotídeo CpG (CpG ODN), nanotubos de carbono (CNTs) e hidróxido de alumínio (Alhydrogel). Formulações contendo rLigAni associada ao polissacarídeo xantana e aos CNTs induziram títulos de anticorpos IgG significativos e comparáveis aos induzidos quando a proteína foi associada ao Alhydrogel. Proteção contra o desafio letal foi observada em 100%, 100%, 67% e 50% dos hamsters imunizados com rLigAni-X1, rLigAni-CpG-X1, rLigAni-Alhydrogel e rLigAni-X2, respectivamente (Fisher test P < 0,05). As preparações contendo rLigAni associada aos CNTs, embora tenham induzido resposta de anticorpos, falharam em conferir imunoproteção. Adicionalmente, os adjuvantes xantana e CNTs não se mostraram tóxicos em células de ovário de hamster Chinês (CHO), in vitro. Os resultados desse estudo apontam a xantana como um novo adjuvante para vacinas de subunidade contra leptospirose, apresentando a propriedade de potencializar a resposta imune contra o antígeno, além de biocompatibilidade e a possibilidade de redução no número de doses requeridas para proteção.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaAdjuvantsXanthanSubunit vaccinesCarbon nanotubesLeptospirosisAdjuvantesXantanaVacinas de subunidadeNanotubos de carbonoLeptospiroseCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIAAdjuvantes: impacto na eficácia de vacina de subunidade contra leptospiroseAdjuvants: impact on the effectiveness in subunit vaccine against leptospirosisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALtese_katia_leston_bacelo.pdfapplication/pdf1389207http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1227/1/tese_katia_leston_bacelo.pdfb5ad0bea30c73a3ecb9d5b196b2a8b64MD51open accessTEXTtese_katia_leston_bacelo.pdf.txttese_katia_leston_bacelo.pdf.txtExtracted Texttext/plain160977http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1227/2/tese_katia_leston_bacelo.pdf.txt7f0d75b5325abdbaf925d7b268cab01bMD52open accessTHUMBNAILtese_katia_leston_bacelo.pdf.jpgtese_katia_leston_bacelo.pdf.jpgGenerated Thumbnailimage/jpeg1784http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1227/3/tese_katia_leston_bacelo.pdf.jpg034e09863bbac01caf38176f8155e638MD53open access123456789/12272018-07-13 09:48:22.372open accessoai:guaiaca.ufpel.edu.br:123456789/1227Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2018-07-13T12:48:22Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false
dc.title.por.fl_str_mv Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
dc.title.alternative.eng.fl_str_mv Adjuvants: impact on the effectiveness in subunit vaccine against leptospirosis
title Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
spellingShingle Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
Bacelo, Kátia Leston
Adjuvants
Xanthan
Subunit vaccines
Carbon nanotubes
Leptospirosis
Adjuvantes
Xantana
Vacinas de subunidade
Nanotubos de carbono
Leptospirose
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA
title_short Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
title_full Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
title_fullStr Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
title_full_unstemmed Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
title_sort Adjuvantes: impacto na eficácia de vacina de subunidade contra leptospirose
author Bacelo, Kátia Leston
author_facet Bacelo, Kátia Leston
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/3849631792846037
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 Bacelo, Kátia Leston
contributor_str_mv Dellagostin, Odir Antônio
dc.subject.eng.fl_str_mv Adjuvants
Xanthan
Subunit vaccines
Carbon nanotubes
Leptospirosis
topic Adjuvants
Xanthan
Subunit vaccines
Carbon nanotubes
Leptospirosis
Adjuvantes
Xantana
Vacinas de subunidade
Nanotubos de carbono
Leptospirose
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA
dc.subject.por.fl_str_mv Adjuvantes
Xantana
Vacinas de subunidade
Nanotubos de carbono
Leptospirose
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA
description A major challenge for the development of vaccines based on purified or recombinant protein subunits, and synthetic peptides resides in the fact that these are poorly immunogenic and mobilize insufficient immunoprotective response. Adjuvants are often used in association with these subunits in order to amplify and direct the immune response induced. Nowadays, there is a wide range of compounds that demonstrate adjuvant activity, however, few are approved for human use, and these often fail to induce appropriate immune response against a particular pathogen. Thus, there is a need to develop new adjuvants that are safe, effective and represent an alternative to currently available. In the present study, we used as a model antigen the non- identical portion of the Leptospira LigA protein (Leptospiral immunoglobulin-like protein A), an outer membrane protein of great interest as a mediator pathogenic mechanisms, used in serological diagnosis and as experimental vaccines. This antigen was formulated with various adjuvants, and the formulations tested for their immunoprotective potential in hamsters, challenged with a virulent strain of L. interrogans serovar Copenhageni. For this, the LigAni protein was produced in recombinant form (rLigAni) using Escherichia coli as the expression system and associated to xanthan polysaccharide, in its variants xanthan pruni strains 106 (X1) and 101 (X2), commercial xanthan and also to oligodinucleotídeo CpG (CpG ODN), carbon nanotubes (CNTs) and aluminum hydroxide (Alhydrogel). Formulations containing rLigAni associated with xanthan polysaccharide and CNTs induced significant IgG antibody titers, comparable to that induced when the protein was associated with Alhydrogel. Protection against lethal challenge was observed in 100%, 100%, 67% and 50% of the hamsters immunized with rLigAni-X1, rLigAni- CpG-X1, rLigAni-Alhydrogel and rLigAni-X2, respectively (Fisher test P < 0.05). The preparations containing rLigAni associated with CNTs, although induced an antibody response, failed to confer immunoprotection. Additionally, xanthan and CNTs adjuvants were not toxic to Chinese hamster ovary (CHO) cells, in vitro. The results of this study indicate xanthan as a new adjuvant for subunit vaccines against leptospirosis, presenting the ability to potentiate the immune response against the antigen, besides biocompatibility and the possibility of reduction of number of doses required for protection.
publishDate 2013
dc.date.issued.fl_str_mv 2013-12-10
dc.date.accessioned.fl_str_mv 2014-08-20T13:32:49Z
dc.date.available.fl_str_mv 2014-01-27
2014-08-20T13:32:49Z
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dc.identifier.citation.fl_str_mv BACELO, Kátia Leston. Adjuvants: impact on the effectiveness in subunit vaccine against leptospirosis. 2013. 111 f. Tese (Doutorado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2013.
dc.identifier.uri.fl_str_mv http://guaiaca.ufpel.edu.br/handle/123456789/1227
identifier_str_mv BACELO, Kátia Leston. Adjuvants: impact on the effectiveness in subunit vaccine against leptospirosis. 2013. 111 f. Tese (Doutorado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2013.
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