Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)

Detalhes bibliográficos
Autor(a) principal: Fischer, Geferson
Data de Publicação: 2007
Tipo de documento: Tese
Idioma: por
Título da fonte: Repositório Institucional da UFPel - Guaiaca
Texto Completo: http://guaiaca.ufpel.edu.br/handle/123456789/1246
Resumo: Vaccine formulation often requires the use of adjuvants which enhance or potentiate humoral and/or cellular responses. Nowadays, many vaccines are formulated with substances such as aluminum hydroxide or oily emulsions, specific for veterinary use. However, the interest on the evaluation of natural substances with adjuvant potential like plant extracts has grown. Propolis produced by honey bees from exudates collected from plants has call the attention of researchers due to several bioactive properties reported, such as antiviral, antiinflamatory and antitumoral action. In addition, although the mechanism of action remains unknown, propolis presents activity on the immune system. The aim of this study was to evaluate the immunomodulator activity of an ethanolic extract of Brazilian green propolis, when used as adjuvant in inactivated vaccines against swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5). The addition of 5 mg/dose of the extract to a vaccine with aluminum hydroxide against SuHV-1 increased the humoral immune response of mice, when compared to the same vaccine without propolis (P<0,01). This effect was more evident when the vaccine was diluted (1:4 and 1:8). The raise in neutralizing antibodies titer, expressed in log2, went from 3 to 4.48 and 2.18 to 4.48, respectively, suggesting that the smaller the antigenic mass or less immunogenic the antigen, more pronounced is the adjuvant effect of propolis. When propolis was used on its own with the antigen, an increase in the titer of neutralizing antibodies determined by seroneutralization was not observed. Besides increasing the humoral immune response, the use of propolis also increased the cellular response, increasing mRNA synthesis of IFN-? in mice splenocytes, measured by RT-PCR. This increase was also observed in the group of animals immunized only with antigen and propolis, unlike the effect observed on the humoral response. The adjuvant effect of propolis was also demonstrated when mice were challenged with 31.6 lethal doses of SuHV-1, 21 days after the second inoculation. The addition of propolis to the vaccine with aluminum hydroxide increased the percentage of protected animals, especially in the higher dilutions, comparing to the vaccine without propolis. Similar result was observed in the group of animals vaccinated only with propolis and antigen. The association of 40 mg/dose of ethanolic extract of green propolis to the oily vaccine against BoHV-5 increased titer of bovine neutrali zing antibodies (P<0,01), when compared to a vaccine without propolis. Thirty days after the second dose the titer went from 35 to 54, and after the third dose it increased from 43 to 67. In addition, there was an increase in the percentage of animals with titer above 32. The inclusion of 20 mg/dose of the extract did not alter the humoral response. The chromatographic analysis of propolis by HPLC showed high levels of phenolic compounds as artepillin C and cynamic acid derived, which may be the main substances with action on the immunologic system. Therefore, green propolis ethanolic extract acted as an adjuvant substance, increasing humoral and cellular immune responses in mice and humoral response in bovines, improving the efficiency of experimental vaccines.
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spelling 2014-08-20T13:32:51Z2007-10-292014-08-20T13:32:51Z2007-02-28FISCHER, Geferson. Evaluation of the immunomodulator activity of green propolis extract in animals vaccinated with swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5). 2007. 90 f. Tese (Doutorado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2007.http://guaiaca.ufpel.edu.br/handle/123456789/1246Vaccine formulation often requires the use of adjuvants which enhance or potentiate humoral and/or cellular responses. Nowadays, many vaccines are formulated with substances such as aluminum hydroxide or oily emulsions, specific for veterinary use. However, the interest on the evaluation of natural substances with adjuvant potential like plant extracts has grown. Propolis produced by honey bees from exudates collected from plants has call the attention of researchers due to several bioactive properties reported, such as antiviral, antiinflamatory and antitumoral action. In addition, although the mechanism of action remains unknown, propolis presents activity on the immune system. The aim of this study was to evaluate the immunomodulator activity of an ethanolic extract of Brazilian green propolis, when used as adjuvant in inactivated vaccines against swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5). The addition of 5 mg/dose of the extract to a vaccine with aluminum hydroxide against SuHV-1 increased the humoral immune response of mice, when compared to the same vaccine without propolis (P<0,01). This effect was more evident when the vaccine was diluted (1:4 and 1:8). The raise in neutralizing antibodies titer, expressed in log2, went from 3 to 4.48 and 2.18 to 4.48, respectively, suggesting that the smaller the antigenic mass or less immunogenic the antigen, more pronounced is the adjuvant effect of propolis. When propolis was used on its own with the antigen, an increase in the titer of neutralizing antibodies determined by seroneutralization was not observed. Besides increasing the humoral immune response, the use of propolis also increased the cellular response, increasing mRNA synthesis of IFN-? in mice splenocytes, measured by RT-PCR. This increase was also observed in the group of animals immunized only with antigen and propolis, unlike the effect observed on the humoral response. The adjuvant effect of propolis was also demonstrated when mice were challenged with 31.6 lethal doses of SuHV-1, 21 days after the second inoculation. The addition of propolis to the vaccine with aluminum hydroxide increased the percentage of protected animals, especially in the higher dilutions, comparing to the vaccine without propolis. Similar result was observed in the group of animals vaccinated only with propolis and antigen. The association of 40 mg/dose of ethanolic extract of green propolis to the oily vaccine against BoHV-5 increased titer of bovine neutrali zing antibodies (P<0,01), when compared to a vaccine without propolis. Thirty days after the second dose the titer went from 35 to 54, and after the third dose it increased from 43 to 67. In addition, there was an increase in the percentage of animals with titer above 32. The inclusion of 20 mg/dose of the extract did not alter the humoral response. The chromatographic analysis of propolis by HPLC showed high levels of phenolic compounds as artepillin C and cynamic acid derived, which may be the main substances with action on the immunologic system. Therefore, green propolis ethanolic extract acted as an adjuvant substance, increasing humoral and cellular immune responses in mice and humoral response in bovines, improving the efficiency of experimental vaccines.A formulação de vacinas frequentemente requer o uso de adjuvantes que prolongam ou potencializam as respostas imunes humoral e/ou celular. Atualmente, muitas vacinas são formuladas a partir de substâncias como o hidróxido de alumínio ou emulsões oleosas, específicas para uso veterinário. Contudo, tem sido crescente o interesse pela avaliação de substâncias naturais com potencial adjuvante, como os extratos derivados de plantas. A própolis, produzida pelas abelhas a partir de exsudatos coletados de plantas, tem despertado o interesse de pesquisadores em função das inúmeras propriedades bioativas relatadas, como ação antiviral, antiinflamatória e antitumoral. Além disso, apesar de desconhecidos muitos dos mecanismos de ação, a própolis apresenta atividade sobre o sistema imune. O objetivo deste estudo foi avaliar a capacidade imunomoduladora de um extrato etanólico da própolis verde brasileira, quando utilizado como adjuvante em vacinas inativadas contra o herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV-5). A adição de 5 mg/dose do extrato a uma vacina com hidróxido de alumínio contra o SuHV-1 incrementou a resposta imune humoral de camundongos, quando comparado com a mesma vacina sem própolis (P<0,01). Este efeito foi mais evidente quando a vacina foi diluída (1:4 e 1:8), podendo-se observar um aumento no título de anticorpos neutralizantes, expresso em log2, que passou de 3 para 4,48 e de 2,18 para 4,48, respectivamente, sugerindo que quanto menor a massa antigênica ou menos imunogênico o antígeno, mais pronunciado é o efeito adjuvante da própolis. Quando a própolis foi utilizada isoladamente com o antígeno, não foi observado aumento no título de anticorpos neutralizantes, determinado por soroneutralização. Além de incrementar a resposta imune humoral, o uso da própolis também aumentou a resposta celular, elevando a síntese de mRNA de IFN-? nos esplenócitos dos camundongos, mensurada pela técnica de RT-PCR. Este aumento foi observado inclusive no grupo de animais imunizados somente com antígeno e própolis, contrariando os resultados da resposta humoral. O efeito adjuvante da própolis foi evidenciado também quando camundongos foram desafiados com 31,6 doses letais do SuHV-1, 21 após a segunda inoculação. A adição da própolis à vacina com hidróxido de alumínio aumentou o percentual de animais protegidos, especialmente nas maiores diluições, em comparação à vacina sem própolis. Resultado semelhante foi observado no grupo de animas vacinados somente com própolis e antígeno. A associação de 40 mg/dose do extrato etanólico de própolis verde à vacina oleosa contra BoHV-5 aumentou o título de anticorpos neutralizantes de bovinos (P<0,01), quando comparado à vacina sem própolis. Trinta dias após a segunda vacinação, o título passou de 35 para 54 e aumentou de 43 para 67, trinta dias após a terceira vacinação. Além disso, houve aumento no percentual de animais com títulos elevados, acima de 32. A inclusão de 20 mg/dose do extrato não alterou a resposta humoral. A análise cromatográfica da própolis por HPLC revelou altos níveis de compostos fenólicos como o artepillin C e derivados do ácido cinâmico, que podem ter sido as principais substâncias com ação sobre o sistema imunológico. Portanto, o extrato etanólico de própolis verde atuou como uma substância adjuvante, incrementando as respostas imunes humoral e celular em camundongos e humoral em bovinos, melhorando a eficiência das vacinas experimentais.application/pdfporUniversidade Federal de PelotasPrograma de Pós-Graduação em BiotecnologiaUFPelBRBiotecnologiaImunologyPropolisImmunological systemImmunomodulationImunologiaPrópolisSistema imunológicoImunomodulaçãoCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)Evaluation of the immunomodulator activity of green propolis extract in animals vaccinated with swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5)info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesishttp://lattes.cnpq.br/6163020760854309http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783392H7Dellagostin, Odir Antôniohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9Leite, Fabio Pereira Leivashttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763076J5Vidor, TelmoFischer, Gefersoninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFPel - Guaiacainstname:Universidade Federal de Pelotas (UFPEL)instacron:UFPELORIGINALtese_geferson_fischer.pdfapplication/pdf1176305http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1246/1/tese_geferson_fischer.pdfaba1e55bca9c3b0b1e42000394239602MD51open accessTEXTtese_geferson_fischer.pdf.txttese_geferson_fischer.pdf.txtExtracted Texttext/plain128025http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1246/2/tese_geferson_fischer.pdf.txt0be263397a3c04a5f20fc8a7e30cf6d8MD52open accessTHUMBNAILtese_geferson_fischer.pdf.jpgtese_geferson_fischer.pdf.jpgGenerated Thumbnailimage/jpeg1727http://guaiaca.ufpel.edu.br/xmlui/bitstream/123456789/1246/3/tese_geferson_fischer.pdf.jpgce07ee15a24f46b3a258c4f37af1d43fMD53open access123456789/12462019-08-23 09:58:43.391open accessoai:guaiaca.ufpel.edu.br:123456789/1246Repositório InstitucionalPUBhttp://repositorio.ufpel.edu.br/oai/requestrippel@ufpel.edu.br || repositorio@ufpel.edu.br || aline.batista@ufpel.edu.bropendoar:2019-08-23T12:58:43Repositório Institucional da UFPel - Guaiaca - Universidade Federal de Pelotas (UFPEL)false
dc.title.por.fl_str_mv Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
dc.title.alternative.eng.fl_str_mv Evaluation of the immunomodulator activity of green propolis extract in animals vaccinated with swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5)
title Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
spellingShingle Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
Fischer, Geferson
Imunology
Propolis
Immunological system
Immunomodulation
Imunologia
Própolis
Sistema imunológico
Imunomodulação
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
title_short Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
title_full Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
title_fullStr Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
title_full_unstemmed Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
title_sort Avaliação da capacidade imunomoduladora de extratos de própolis verde em animais vacinados com herpesvírus suíno tipo 1 (SuHV-1) ou herpesvírus bovino tipo 5 (BoHV -5)
author Fischer, Geferson
author_facet Fischer, Geferson
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/6163020760854309
dc.contributor.advisorLattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783392H7
dc.contributor.referees1.pt_BR.fl_str_mv Dellagostin, Odir Antônio
dc.contributor.referees1ID.por.fl_str_mv
dc.contributor.referees1Lattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723107D9
dc.contributor.referees2.pt_BR.fl_str_mv Leite, Fabio Pereira Leivas
dc.contributor.referees2ID.por.fl_str_mv
dc.contributor.referees2Lattes.por.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4763076J5
dc.contributor.advisor1.fl_str_mv Vidor, Telmo
dc.contributor.author.fl_str_mv Fischer, Geferson
contributor_str_mv Vidor, Telmo
dc.subject.eng.fl_str_mv Imunology
Propolis
Immunological system
Immunomodulation
topic Imunology
Propolis
Immunological system
Immunomodulation
Imunologia
Própolis
Sistema imunológico
Imunomodulação
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
dc.subject.por.fl_str_mv Imunologia
Própolis
Sistema imunológico
Imunomodulação
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::MEDICINA VETERINARIA PREVENTIVA::SAUDE ANIMAL (PROGRAMAS SANITARIOS)
description Vaccine formulation often requires the use of adjuvants which enhance or potentiate humoral and/or cellular responses. Nowadays, many vaccines are formulated with substances such as aluminum hydroxide or oily emulsions, specific for veterinary use. However, the interest on the evaluation of natural substances with adjuvant potential like plant extracts has grown. Propolis produced by honey bees from exudates collected from plants has call the attention of researchers due to several bioactive properties reported, such as antiviral, antiinflamatory and antitumoral action. In addition, although the mechanism of action remains unknown, propolis presents activity on the immune system. The aim of this study was to evaluate the immunomodulator activity of an ethanolic extract of Brazilian green propolis, when used as adjuvant in inactivated vaccines against swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5). The addition of 5 mg/dose of the extract to a vaccine with aluminum hydroxide against SuHV-1 increased the humoral immune response of mice, when compared to the same vaccine without propolis (P<0,01). This effect was more evident when the vaccine was diluted (1:4 and 1:8). The raise in neutralizing antibodies titer, expressed in log2, went from 3 to 4.48 and 2.18 to 4.48, respectively, suggesting that the smaller the antigenic mass or less immunogenic the antigen, more pronounced is the adjuvant effect of propolis. When propolis was used on its own with the antigen, an increase in the titer of neutralizing antibodies determined by seroneutralization was not observed. Besides increasing the humoral immune response, the use of propolis also increased the cellular response, increasing mRNA synthesis of IFN-? in mice splenocytes, measured by RT-PCR. This increase was also observed in the group of animals immunized only with antigen and propolis, unlike the effect observed on the humoral response. The adjuvant effect of propolis was also demonstrated when mice were challenged with 31.6 lethal doses of SuHV-1, 21 days after the second inoculation. The addition of propolis to the vaccine with aluminum hydroxide increased the percentage of protected animals, especially in the higher dilutions, comparing to the vaccine without propolis. Similar result was observed in the group of animals vaccinated only with propolis and antigen. The association of 40 mg/dose of ethanolic extract of green propolis to the oily vaccine against BoHV-5 increased titer of bovine neutrali zing antibodies (P<0,01), when compared to a vaccine without propolis. Thirty days after the second dose the titer went from 35 to 54, and after the third dose it increased from 43 to 67. In addition, there was an increase in the percentage of animals with titer above 32. The inclusion of 20 mg/dose of the extract did not alter the humoral response. The chromatographic analysis of propolis by HPLC showed high levels of phenolic compounds as artepillin C and cynamic acid derived, which may be the main substances with action on the immunologic system. Therefore, green propolis ethanolic extract acted as an adjuvant substance, increasing humoral and cellular immune responses in mice and humoral response in bovines, improving the efficiency of experimental vaccines.
publishDate 2007
dc.date.available.fl_str_mv 2007-10-29
2014-08-20T13:32:51Z
dc.date.issued.fl_str_mv 2007-02-28
dc.date.accessioned.fl_str_mv 2014-08-20T13:32:51Z
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dc.identifier.citation.fl_str_mv FISCHER, Geferson. Evaluation of the immunomodulator activity of green propolis extract in animals vaccinated with swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5). 2007. 90 f. Tese (Doutorado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2007.
dc.identifier.uri.fl_str_mv http://guaiaca.ufpel.edu.br/handle/123456789/1246
identifier_str_mv FISCHER, Geferson. Evaluation of the immunomodulator activity of green propolis extract in animals vaccinated with swine herpesvirus type 1 (SuHV-1) or bovine herpesvirus type 5 (BoHV-5). 2007. 90 f. Tese (Doutorado em Biotecnologia) - Universidade Federal de Pelotas, Pelotas, 2007.
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