Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus

Detalhes bibliográficos
Autor(a) principal: Stegmiller, Nataly Pescinalli
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)
Texto Completo: http://repositorio.ufes.br/handle/10/1893
Resumo: Mastitis is a disease of the mammary gland characterized by an inflammatory process, often due to the infectious microorganisms presence. It is considered the main cause of economic losses related to dairy practice, as it can act directly in the organ function, decreasing to 15-20% milk/animal production. Among the microorganisms identified as causing the mastitis, Staphylococcus aureus bacteria species are the most common, accounting for over 91% of mastitis cases. The emergence of drug-resistant strains associated with high toxicity of the treatment, prints the need to develop an effective, inexpensive and safe vaccine that could be used as a preventive measure against mastitis. In this work, we aimed to develop a vaccine produced with total antigen of S. aureus (SaAg) and intranasaly or intramuscularly given against experimental mastitis. The vaccine preparation was done using a S. aureus strain isolated from bovine mammary gland, followed of 15 slow freezing /thawing cycles. BALB/c mice were vaccinated by intranasal or intramuscular route with 2 doses of SaAg (50μg/animal) followed of biocompatibility and immunogenicity evaluation 72 hours after each dose. Intramuscular vaccinated mice showed behavioral and physical changes. No differences of transaminases and creatinine levels were observed in the vaccinated groups as compared to unvaccinated animals, different from observed in the lymphoproliferative responses. Intranasal vaccinated mice presented significant percentage increase of CD4+ and CD8+ IFN-γ producer’s when compared to intramuscular or unvaccinated groups. By the other hand, mice vaccinated by intramuscular route showed a increase of CD4+ and CD8+ T cells producers percentage also supernatant cytokine production (Il-10 and IFN- γ) after SaAg in vitro recall, when compared to intramuscular vaccinated animals or unvaccinated. The Intranasal vaccination induced a significant increase of IgG2a levels and decreased IgG1 production when compared to control or intramuscular vaccinated groups. Thus, our results demonstrate the feasibility of SaAg vaccine, given by intramuscular and intranasal routes, as a potential prophylactic agent against mastitis caused by S. aureus.
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spelling Guimarães, Marco César CunegundesGomes, Daniel Cláudio de OliveiraStegmiller, Nataly PescinalliNogueira, Breno ValentimBergmann, Bartira RossiPereira, Fausto Edmundo Lima2016-05-17T16:02:43Z2016-06-24T06:00:05Z2014-04-302014-04-30Mastitis is a disease of the mammary gland characterized by an inflammatory process, often due to the infectious microorganisms presence. It is considered the main cause of economic losses related to dairy practice, as it can act directly in the organ function, decreasing to 15-20% milk/animal production. Among the microorganisms identified as causing the mastitis, Staphylococcus aureus bacteria species are the most common, accounting for over 91% of mastitis cases. The emergence of drug-resistant strains associated with high toxicity of the treatment, prints the need to develop an effective, inexpensive and safe vaccine that could be used as a preventive measure against mastitis. In this work, we aimed to develop a vaccine produced with total antigen of S. aureus (SaAg) and intranasaly or intramuscularly given against experimental mastitis. The vaccine preparation was done using a S. aureus strain isolated from bovine mammary gland, followed of 15 slow freezing /thawing cycles. BALB/c mice were vaccinated by intranasal or intramuscular route with 2 doses of SaAg (50μg/animal) followed of biocompatibility and immunogenicity evaluation 72 hours after each dose. Intramuscular vaccinated mice showed behavioral and physical changes. No differences of transaminases and creatinine levels were observed in the vaccinated groups as compared to unvaccinated animals, different from observed in the lymphoproliferative responses. Intranasal vaccinated mice presented significant percentage increase of CD4+ and CD8+ IFN-γ producer’s when compared to intramuscular or unvaccinated groups. By the other hand, mice vaccinated by intramuscular route showed a increase of CD4+ and CD8+ T cells producers percentage also supernatant cytokine production (Il-10 and IFN- γ) after SaAg in vitro recall, when compared to intramuscular vaccinated animals or unvaccinated. The Intranasal vaccination induced a significant increase of IgG2a levels and decreased IgG1 production when compared to control or intramuscular vaccinated groups. Thus, our results demonstrate the feasibility of SaAg vaccine, given by intramuscular and intranasal routes, as a potential prophylactic agent against mastitis caused by S. aureus.A mastite é uma enfermidade da glândula mamária que se caracteriza por um processo inflamatório, quase sempre decorrente da presença de microrganismos infecciosos. É considerada a principal causa de perdas econômicas relacionadas à prática leiteira, já que interfere diretamente na função do órgão, diminuindo de 15-20% a produção láctea/animal. Dentre os microrganismos identificados como causadores da mastite, bactérias da espécie Staphylococcus aureus são as mais comumente relacionadas, sendo responsáveis por mais de 91% dos casos. O surgimento de cepas droga-resistentes associado a grande toxicidade do tratamento, imprime a necessidade do desenvolvimento de uma vacina efetiva, segura e barata, que possa ser utilizada como medida preventiva contra a mastite. Por esse motivo, objetivamos o desenvolvimento de uma vacina composta por antígenos totais de S. aureus (SaAg), administrada por via intranasal ou intramuscular, contra a mastite experimental. O preparo da vacina foi realizado a partir da cultura de uma cepa de S. aureus isolada de glândula mamária bovina, caracterizada molecularmente e submetida a 15 ciclos de congelamento e descongelamento lento. Camundongos BALB/c, foram vacinados pelas vias intranasal ou intramuscular com SaAg (50 g/mL) em 2 doses intervaladas por 15 dias, para avaliação da biocompatibilidade e imunogenicidade. Os sinais clínicos foram monitorados por 72 horas após cada dose, onde se observou alterações comportamentais e físicas, com intensa resposta inflamatória nos animais vacinados por via intramuscular. Não foram observadas diferenças nos níveis de transaminases e creatinina entre os grupos vacinados por via nasal ou intramuscular em comparação com o controle. Além disso, foi verificado um aumento significativo das respostas linfoproliferativas dos animais vacinados em relação aos controles. Animais vacinados por via intranasal apresentaram maiores porcentagens de linfócitos TCD4+ e TCD8+ produtores de IFN-γ quando comparados aos demais grupos, o que não foi verificado na avaliação da produção de citocinas utilizando o sobrenadante das culuras estimuladas com SaAg in vitro. De forma diferente, animais vacinados por via intramuscular apresentaram maiores porcentagens de linfócitos TCD4+ e TCD8+ produtores de IL-10, quando comparados aos demais grupos, bem como produziram maiores níveis de IFN-γ nos sobrenadantes de cultura reestimulados, quando comparados aos grupos controle ou vacinados por via intranasal. Na avaliação da resposta imune humoral, significativos níveis de IgG2a foram relacionados à vacinação intranasal, quando comparados aos demais grupos. Além disso, animais vacinados por via intranasal ou intramuscular apresentaram semelhantes níveis de IgG1, porém, significativamente maiores que os animais controle. Dessa forma, nossos resultados demonstram a viabilidade do uso da vacina SaAg, administrada pelas vias intramuscular e intranasal, como um potencial agente profilático contra a mastite causada por S. aureus.CAPESTexthttp://repositorio.ufes.br/handle/10/1893porUniversidade Federal do Espírito SantoMestrado em BiotecnologiaPrograma de Pós-Graduação em BiotecnologiaUFESBRCentro de Ciências da SaúdeVaccineMastitisImmune responseStaphylococcus aureusMastiteVacinasResposta imuneBiotecnologia61Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureusinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da Universidade Federal do Espírito Santo (riUfes)instname:Universidade Federal do Espírito Santo (UFES)instacron:UFESCAPESORIGINALDissertacao Nataly Stegmiller.pdfDissertacao Nataly Stegmiller.pdfapplication/pdf969971http://repositorio.ufes.br/bitstreams/86d357ef-8058-44e0-a79d-b9e29fe72b43/download56211b8da0204ded63a6d90ae86998e6MD51CC-LICENSElicense_urllicense_urltext/plain; charset=utf-849http://repositorio.ufes.br/bitstreams/f6dc0459-06f9-4e62-9be9-c742c7c5c23f/download4afdbb8c545fd630ea7db775da747b2fMD52license_textlicense_texttext/html; charset=utf-822064http://repositorio.ufes.br/bitstreams/0ef40748-ad30-4cf9-960c-a28a14ec845d/downloadef48816a10f2d45f2e2fee2f478e2fafMD53license_rdflicense_rdfapplication/rdf+xml; charset=utf-823148http://repositorio.ufes.br/bitstreams/456ca857-f55e-4de6-befc-fac9731733c3/download9da0b6dfac957114c6a7714714b86306MD54LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufes.br/bitstreams/3874b08e-5d42-4d5c-b37a-768ab01ce47b/download8a4605be74aa9ea9d79846c1fba20a33MD5510/18932024-06-27 11:06:52.962oai:repositorio.ufes.br:10/1893http://repositorio.ufes.brRepositório InstitucionalPUBhttp://repositorio.ufes.br/oai/requestopendoar:21082024-06-27T11:06:52Repositório Institucional da Universidade Federal do Espírito Santo (riUfes) - Universidade Federal do Espírito Santo (UFES)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
dc.title.none.fl_str_mv Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
title Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
spellingShingle Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
Stegmiller, Nataly Pescinalli
Vaccine
Mastitis
Immune response
Biotecnologia
Staphylococcus aureus
Mastite
Vacinas
Resposta imune
61
title_short Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
title_full Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
title_fullStr Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
title_full_unstemmed Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
title_sort Comparação entre as vias intranasal e intramuscular para a imunização com uma vacina de antígeno bruto de Staphylococcus aureus
author Stegmiller, Nataly Pescinalli
author_facet Stegmiller, Nataly Pescinalli
author_role author
dc.contributor.advisor-co1.fl_str_mv Guimarães, Marco César Cunegundes
dc.contributor.advisor1.fl_str_mv Gomes, Daniel Cláudio de Oliveira
dc.contributor.author.fl_str_mv Stegmiller, Nataly Pescinalli
dc.contributor.referee1.fl_str_mv Nogueira, Breno Valentim
dc.contributor.referee2.fl_str_mv Bergmann, Bartira Rossi
dc.contributor.referee3.fl_str_mv Pereira, Fausto Edmundo Lima
contributor_str_mv Guimarães, Marco César Cunegundes
Gomes, Daniel Cláudio de Oliveira
Nogueira, Breno Valentim
Bergmann, Bartira Rossi
Pereira, Fausto Edmundo Lima
dc.subject.eng.fl_str_mv Vaccine
Mastitis
Immune response
topic Vaccine
Mastitis
Immune response
Biotecnologia
Staphylococcus aureus
Mastite
Vacinas
Resposta imune
61
dc.subject.cnpq.fl_str_mv Biotecnologia
dc.subject.br-rjbn.none.fl_str_mv Staphylococcus aureus
Mastite
Vacinas
Resposta imune
dc.subject.udc.none.fl_str_mv 61
description Mastitis is a disease of the mammary gland characterized by an inflammatory process, often due to the infectious microorganisms presence. It is considered the main cause of economic losses related to dairy practice, as it can act directly in the organ function, decreasing to 15-20% milk/animal production. Among the microorganisms identified as causing the mastitis, Staphylococcus aureus bacteria species are the most common, accounting for over 91% of mastitis cases. The emergence of drug-resistant strains associated with high toxicity of the treatment, prints the need to develop an effective, inexpensive and safe vaccine that could be used as a preventive measure against mastitis. In this work, we aimed to develop a vaccine produced with total antigen of S. aureus (SaAg) and intranasaly or intramuscularly given against experimental mastitis. The vaccine preparation was done using a S. aureus strain isolated from bovine mammary gland, followed of 15 slow freezing /thawing cycles. BALB/c mice were vaccinated by intranasal or intramuscular route with 2 doses of SaAg (50μg/animal) followed of biocompatibility and immunogenicity evaluation 72 hours after each dose. Intramuscular vaccinated mice showed behavioral and physical changes. No differences of transaminases and creatinine levels were observed in the vaccinated groups as compared to unvaccinated animals, different from observed in the lymphoproliferative responses. Intranasal vaccinated mice presented significant percentage increase of CD4+ and CD8+ IFN-γ producer’s when compared to intramuscular or unvaccinated groups. By the other hand, mice vaccinated by intramuscular route showed a increase of CD4+ and CD8+ T cells producers percentage also supernatant cytokine production (Il-10 and IFN- γ) after SaAg in vitro recall, when compared to intramuscular vaccinated animals or unvaccinated. The Intranasal vaccination induced a significant increase of IgG2a levels and decreased IgG1 production when compared to control or intramuscular vaccinated groups. Thus, our results demonstrate the feasibility of SaAg vaccine, given by intramuscular and intranasal routes, as a potential prophylactic agent against mastitis caused by S. aureus.
publishDate 2014
dc.date.submitted.none.fl_str_mv 2014-04-30
dc.date.issued.fl_str_mv 2014-04-30
dc.date.accessioned.fl_str_mv 2016-05-17T16:02:43Z
dc.date.available.fl_str_mv 2016-06-24T06:00:05Z
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dc.publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Mestrado em Biotecnologia
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dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Centro de Ciências da Saúde
publisher.none.fl_str_mv Universidade Federal do Espírito Santo
Mestrado em Biotecnologia
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