Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração
में बचाया:
| मुख्य लेखक: | |
|---|---|
| प्रकाशन तिथि: | 2017 |
| स्वरूप: | Master thesis |
| भाषा: | por |
| स्रोत: | Repositório Institucional da UPF |
| Download full: | https://repositorio.upf.br/handle/123456789/1596 |
सारांश: | Gonadotrophin releasing hormone (GnRH) has as its main function regulate the reproduction of vertebrates. Active immunization against GnRH has been used strategically to neutralize endogenous GnRH and to promote immunocastration of vaccinated animals. This method replaces surgically castrated and increases the quality of life of the animals. In this work, we present the development and evaluation of two experimental vaccines based on swine GnRH with the potential to neutralize endogenous GnRH in rats. The genes encoding the recombinant proteins Construct007 (carrier 001 + GnRH) and constructCTBsGnRh7-24His (carrier 002 + GnRH) were synthesized by Amplicon Vaccine (Pullman, USA) and cloned into the expression vector pET24. Thereafter, the constructs were chemically transformed into Escherichia coli strain ER2566 and the expression of the recombinant proteins was induced overnight with IPTG. After cell lysis, the proteins were purified by liquid protein chromatography using a column loaded with nickel ions. After dialysis of the proteins in PBS (pH 7.2), the experimental vaccines were formulated with 50 μg of the pure antigens potentiated with 15% Montanide Gel 01 adjuvant (Seppic, France). A total of 12 male adult Wistar rats were randomly separated into 3 groups. The first two groups were immunized intramuscularly with either the Construct007 or constructCTBsGnRh7-24His vaccine. The third group was inoculated with PBS + Montanide Gel 01. The vaccination protocol consisted of 2 doses with a 21 day interval. Clinically, the safety of formulations and testicular development (testis size and weight), and immunologically the ability of vaccines to induce anti-GnRH pig antibodies were evaluated. Animals immunized with Construct007-based vaccine showed significantly higher antibody titers compared to those vaccinated with recombinant constructCTBsGnRh7-24His protein. Clinically, the antibody response induced by the Construct007 vaccine was able to neutralize the endogenous GnRH of rats, significantly reducing the testicular development of these animals compared to control animals (PBS + Montanide Gel 01), as well, compared to those vaccinated with the constructCTBsGnRh7-24His protein. Taking into account the safety and functional blocking ability of GnRH induced by the Construct007 formulation, it becomes evident the potential use of this experimental vaccine in a future study in the immunization target species. This dissertation contains data that are potentially patentable and should be kept confidential until 13 xv they are processed by patent and international regulatory agencies. Key words: GnRH, Vaccines, Swine, immunocastration. |
| _version_ | 1871607636353351680 |
|---|---|
| author | Andreetta, Angélica Teresinha |
| author_browse | Andreetta, Angélica Teresinha |
| author_facet | Andreetta, Angélica Teresinha |
| author_role | author |
| collection | Repositório Institucional da UPF |
| dc.contributor.author.fl_str_mv | Andreetta, Angélica Teresinha |
| dc.contributor.none.fl_str_mv | Zanella, Eraldo Lourenso lattes.cnpq.br/9221953442964161 |
| dc.date.none.fl_str_mv | 2017-09-29 2025-05-07T00:27:08Z |
| dc.format.none.fl_str_mv | application/pdf application/pdf |
| dc.identifier.uri.fl_str_mv | ANDREETTA, Angélica Teresinha. Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração. 2017. 48 f. Dissertação (Mestrado em Bioexperimentação) - Universidade de Passo Fundo, Passo Fundo, RS, 2017. https://repositorio.upf.br/handle/123456789/1596 |
| dc.language.iso.fl_str_mv | por |
| dc.publisher.none.fl_str_mv | Universidade de Passo Fundo Faculdade de Agronomia e Medicina Veterinária – FAMV Brasil UPF Programa de Pós-Graduação em Bioexperimentação |
| dc.rights.driver.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.source.none.fl_str_mv | reponame:Repositório Institucional da UPF instname:Universidade de Passo Fundo (UPF) instacron:UPF |
| dc.subject.por.fl_str_mv | Suíno Vacinas Castração CIENCIA E TECNOLOGIA DE ALIMENTOS::ENGENHARIA DE ALIMENTOS |
| dc.title.none.fl_str_mv | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração Development and immunogenic evaluation of two vaccines based on swine GnRH with potential use in immunocastration |
| dc.type.driver.fl_str_mv | info:eu-repo/semantics/masterThesis |
| dc.type.status.fl_str_mv | info:eu-repo/semantics/publishedVersion |
| description | Gonadotrophin releasing hormone (GnRH) has as its main function regulate the reproduction of vertebrates. Active immunization against GnRH has been used strategically to neutralize endogenous GnRH and to promote immunocastration of vaccinated animals. This method replaces surgically castrated and increases the quality of life of the animals. In this work, we present the development and evaluation of two experimental vaccines based on swine GnRH with the potential to neutralize endogenous GnRH in rats. The genes encoding the recombinant proteins Construct007 (carrier 001 + GnRH) and constructCTBsGnRh7-24His (carrier 002 + GnRH) were synthesized by Amplicon Vaccine (Pullman, USA) and cloned into the expression vector pET24. Thereafter, the constructs were chemically transformed into Escherichia coli strain ER2566 and the expression of the recombinant proteins was induced overnight with IPTG. After cell lysis, the proteins were purified by liquid protein chromatography using a column loaded with nickel ions. After dialysis of the proteins in PBS (pH 7.2), the experimental vaccines were formulated with 50 μg of the pure antigens potentiated with 15% Montanide Gel 01 adjuvant (Seppic, France). A total of 12 male adult Wistar rats were randomly separated into 3 groups. The first two groups were immunized intramuscularly with either the Construct007 or constructCTBsGnRh7-24His vaccine. The third group was inoculated with PBS + Montanide Gel 01. The vaccination protocol consisted of 2 doses with a 21 day interval. Clinically, the safety of formulations and testicular development (testis size and weight), and immunologically the ability of vaccines to induce anti-GnRH pig antibodies were evaluated. Animals immunized with Construct007-based vaccine showed significantly higher antibody titers compared to those vaccinated with recombinant constructCTBsGnRh7-24His protein. Clinically, the antibody response induced by the Construct007 vaccine was able to neutralize the endogenous GnRH of rats, significantly reducing the testicular development of these animals compared to control animals (PBS + Montanide Gel 01), as well, compared to those vaccinated with the constructCTBsGnRh7-24His protein. Taking into account the safety and functional blocking ability of GnRH induced by the Construct007 formulation, it becomes evident the potential use of this experimental vaccine in a future study in the immunization target species. This dissertation contains data that are potentially patentable and should be kept confidential until 13 xv they are processed by patent and international regulatory agencies. Key words: GnRH, Vaccines, Swine, immunocastration. |
| eu_rights_str_mv | openAccess |
| format | masterThesis |
| id | UPF_8a45cd2fcb80aee07a3079d9674f4481 |
| identifier_str_mv | ANDREETTA, Angélica Teresinha. Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração. 2017. 48 f. Dissertação (Mestrado em Bioexperimentação) - Universidade de Passo Fundo, Passo Fundo, RS, 2017. |
| instacron_str | UPF |
| institution | UPF |
| instname_str | Universidade de Passo Fundo (UPF) |
| language | por |
| network_acronym_str | UPF |
| network_name_str | Repositório Institucional da UPF |
| oai_identifier_str | oai:repositorio.upf.br:123456789/1596 |
| publishDate | 2017 |
| publishDateSort | 2017 |
| publisher.none.fl_str_mv | Universidade de Passo Fundo Faculdade de Agronomia e Medicina Veterinária – FAMV Brasil UPF Programa de Pós-Graduação em Bioexperimentação |
| reponame_str | Repositório Institucional da UPF |
| repository.mail.fl_str_mv | jucelei@upf.br||biblio@upf.br |
| repository.name.fl_str_mv | Repositório Institucional da UPF - Universidade de Passo Fundo (UPF) |
| repository_id_str | 1610 |
| spelling | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastraçãoDevelopment and immunogenic evaluation of two vaccines based on swine GnRH with potential use in immunocastrationSuínoVacinasCastraçãoCIENCIA E TECNOLOGIA DE ALIMENTOS::ENGENHARIA DE ALIMENTOSGonadotrophin releasing hormone (GnRH) has as its main function regulate the reproduction of vertebrates. Active immunization against GnRH has been used strategically to neutralize endogenous GnRH and to promote immunocastration of vaccinated animals. This method replaces surgically castrated and increases the quality of life of the animals. In this work, we present the development and evaluation of two experimental vaccines based on swine GnRH with the potential to neutralize endogenous GnRH in rats. The genes encoding the recombinant proteins Construct007 (carrier 001 + GnRH) and constructCTBsGnRh7-24His (carrier 002 + GnRH) were synthesized by Amplicon Vaccine (Pullman, USA) and cloned into the expression vector pET24. Thereafter, the constructs were chemically transformed into Escherichia coli strain ER2566 and the expression of the recombinant proteins was induced overnight with IPTG. After cell lysis, the proteins were purified by liquid protein chromatography using a column loaded with nickel ions. After dialysis of the proteins in PBS (pH 7.2), the experimental vaccines were formulated with 50 μg of the pure antigens potentiated with 15% Montanide Gel 01 adjuvant (Seppic, France). A total of 12 male adult Wistar rats were randomly separated into 3 groups. The first two groups were immunized intramuscularly with either the Construct007 or constructCTBsGnRh7-24His vaccine. The third group was inoculated with PBS + Montanide Gel 01. The vaccination protocol consisted of 2 doses with a 21 day interval. Clinically, the safety of formulations and testicular development (testis size and weight), and immunologically the ability of vaccines to induce anti-GnRH pig antibodies were evaluated. Animals immunized with Construct007-based vaccine showed significantly higher antibody titers compared to those vaccinated with recombinant constructCTBsGnRh7-24His protein. Clinically, the antibody response induced by the Construct007 vaccine was able to neutralize the endogenous GnRH of rats, significantly reducing the testicular development of these animals compared to control animals (PBS + Montanide Gel 01), as well, compared to those vaccinated with the constructCTBsGnRh7-24His protein. Taking into account the safety and functional blocking ability of GnRH induced by the Construct007 formulation, it becomes evident the potential use of this experimental vaccine in a future study in the immunization target species. This dissertation contains data that are potentially patentable and should be kept confidential until 13 xv they are processed by patent and international regulatory agencies. Key words: GnRH, Vaccines, Swine, immunocastration.O hormônio liberador de gonadotrofina (GnRH) tem como principal função atuar na regulação da reprodução de vertebrados. A imunização ativa contra o GnRH tem sido utilizada estrategicamente para neutralizar o GnRH endógeno e promover a imunocastração dos animais vacinados. Este método substitui a castração cirúrgica e aumenta a qualidade de vida dos animais. Neste trabalho, apresentamos o desenvolvimento e a avaliação de duas vacinas experimentais baseadas no GnRH suíno com potencial de neutralização do GnRH endógeno de ratos. Os genes codificantes das proteínas recombinantes Construct007 (carreador 001 + GnRH) e constructCTBsGnRh7-24His (carreador 002 + GnRH) foram sintetizados pela empresa Amplicon Vaccine (Pullman, USA) e clonados no vetor de expressão pET24. Em seguida, as construções foram transformadas quimicamente em Escherichia coli cepa ER2566 e a expressão das proteínas recombinantes foi induzida overnight com IPTG. Após lise celular, procedeu-se a purificação das proteínas mediante cromatrografia líquida de proteína utilizando uma coluna carregada com íons de níquel. Após a diálise das proteínas em PBS (pH 7,2), as vacinas experimentais foram formuladas com 50 ¿g dos antígenos puros potencializados com 15% de adjuvante Montanide Gel 01 (Seppic, França). Um total de 12 ratos Wistar, machos adultos, foram aleatoriamente separados em 3 grupos. Os dois primeiros grupos foram imunizados pela via intramuscular com a vacina Construct007 ou constructCTBsGnRh7-24His. O terceiro grupo foi inoculado com PBS + Montanide Gel 01. O protocolo de vacinação consistiu em 2 doses com intervalo de 21 dias. Clinicamente, avaliou-se a inocuidade das formulações e o desenvolvimento testicular (tamanho e peso dos testículos), e imunologicamente a capacidade das vacinas de induzirem anticorpos anti GnRH suíno. Os animais imunizados com vacina baseada na proteína Construct007 apresentaram títulos de anticorpos significativamente superiores em comparação com aqueles que foram vacinados com proteína recombinante constructCTBsGnRh7-24His. Clinicamente, a resposta de anticorpos induzida pela vacina Construct007 foi capaz de neutralizar o GnRH endógeno de ratos, reduzindo significativamente o desenvolvimento testicular destes animais em comparação com os animais controles (PBS + Montanide Gel 01), bem como, em comparação com aqueles vacinados com a 11 xii i proteína constructCTBsGnRh7-24His. Levando em consideração a inocuidade e a capacidade funcional de bloqueio do GnRH induzida pela formulação Construct007, torna-se evidente o potencial uso desta vacina experimental em um futuro estudo de imunização na espécie alvo. Esta dissertação contém dados potencialmente patenteáveis e que devem ser mantidos em sigilo até sua tramitação junto as agências regulatórias de patentes nacionais e internacionais.Universidade de Passo FundoFaculdade de Agronomia e Medicina Veterinária – FAMVBrasilUPFPrograma de Pós-Graduação em BioexperimentaçãoZanella, Eraldo Lourensolattes.cnpq.br/9221953442964161Andreetta, Angélica Teresinha2025-05-07T00:27:08Z2017-09-29info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfapplication/pdfANDREETTA, Angélica Teresinha. Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração. 2017. 48 f. Dissertação (Mestrado em Bioexperimentação) - Universidade de Passo Fundo, Passo Fundo, RS, 2017.https://repositorio.upf.br/handle/123456789/1596porinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UPFinstname:Universidade de Passo Fundo (UPF)instacron:UPF2025-12-30T19:54:32Zoai:repositorio.upf.br:123456789/1596Repositório InstitucionalPRIhttp://repositorio.upf.br/oai/requestjucelei@upf.br||biblio@upf.bropendoar:16102025-12-30T19:54:32Repositório Institucional da UPF - Universidade de Passo Fundo (UPF) |
| spellingShingle | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração Andreetta, Angélica Teresinha Suíno Vacinas Castração CIENCIA E TECNOLOGIA DE ALIMENTOS::ENGENHARIA DE ALIMENTOS |
| status_str | publishedVersion |
| title | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração |
| title_full | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração |
| title_fullStr | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração |
| title_full_unstemmed | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração |
| title_short | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração |
| title_sort | Desenvolvimento e avaliação imunogênica de duas vacinas baseadas no GnRH suíno com potencial uso em imunocastração |
| topic | Suíno Vacinas Castração CIENCIA E TECNOLOGIA DE ALIMENTOS::ENGENHARIA DE ALIMENTOS |
| url | https://repositorio.upf.br/handle/123456789/1596 |