Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos
Autor(a) principal: | |
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Data de Publicação: | 2004 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/27142 http://dx.doi.org/10.14393/ufu.di.2004.28 |
Resumo: | Paracoccidioides brasiliensis has antigenic components on surface able to promote the interaction between fungus and host, and promote the infection. In the present study the presence of JBE (Jacalin Binding Exoantigen) in culture supematant of BAT P. brasiliensis isolate and on yeast cells were verified and the effect of JBE in attachment to murine peritoneal macrophages was investigated. JBE was obtained in yeast cells culture supematant (liquid F-10 médium containing 0.5 % D-Glucose) and showed by SDS-PAGE and silver nitrate staining a 190 kDa component and, under reducing conditions, a 70 kDa component. Moreover, JBE was localized by rabbit anti-gpl90 IgG and immunogold reaction on yeast cells surface. Peritoneal macrophages form BALB/c mice were treated with JBE (50 e 100 pg/mL) and incubated with P. brasiliensis yeast cells FITC labelling pre-treated with 50 mM monosaccharides: N-acetyl-D-glucosamine (GlcNAc), or D-Glucose, or D-Galactose or D-Mannose. JBE was able to promote phagocytosis inhibition (40 %) and NO release by JBE-treated macrophages. The phagocytosis inhibition increased to 64 % when JBE-treated macrophages were incubated with yeast cells pre-treated with GlcNAc, though NO release was decreased. Yeast cells were treated with mice anti-JBE IgG F(ab) polyclonal fragments (50 e 100 pg/mL) and macrophages treated with 50 mM monosaccharides promoted phagocytosis inhibition, the incubation of both promoted a increase on phagocytosis inhibition. The incubation of JBE-treated macrophages (50 pg/mL) and yeast cells pre-treated with anti-JBE IgG F(ab) (50 pg/mL) promoted phagocytosis inhibition (61,5 %). This results suggest that JBE could be involved on the fungus and host interaction because its localization. Furthermore, JBE potentializes the phagocytosis of P. brasiliensis yeast cells by murine peritoneal macrophages. Probably, the phagocytosis inhibition and NO decrease by GlcNAc is due to gp 70 affinity to GlcNAc. The anti-JBE IgG F(ab) probably occupies sites of interaction on yeast cells and improve the phagocytosis inhibition by monosaccharides or JBE on macrophages. In conclusion, JBE has component able to promote invasion of fungus. |
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Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinosEvaluation of JBE (“Jacalin Binding Exoantigen”) in phagocytosis of Paracoccidioides brasiliensis yeast by murine peritoneal macrophagesParacoccidioides brasiliensisMacrófagosExoantígenoCarboidratoParacoccidioides brasiliensisMacrophagesExoantigenCarbohydratesCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIAParacoccidioides brasiliensis has antigenic components on surface able to promote the interaction between fungus and host, and promote the infection. In the present study the presence of JBE (Jacalin Binding Exoantigen) in culture supematant of BAT P. brasiliensis isolate and on yeast cells were verified and the effect of JBE in attachment to murine peritoneal macrophages was investigated. JBE was obtained in yeast cells culture supematant (liquid F-10 médium containing 0.5 % D-Glucose) and showed by SDS-PAGE and silver nitrate staining a 190 kDa component and, under reducing conditions, a 70 kDa component. Moreover, JBE was localized by rabbit anti-gpl90 IgG and immunogold reaction on yeast cells surface. Peritoneal macrophages form BALB/c mice were treated with JBE (50 e 100 pg/mL) and incubated with P. brasiliensis yeast cells FITC labelling pre-treated with 50 mM monosaccharides: N-acetyl-D-glucosamine (GlcNAc), or D-Glucose, or D-Galactose or D-Mannose. JBE was able to promote phagocytosis inhibition (40 %) and NO release by JBE-treated macrophages. The phagocytosis inhibition increased to 64 % when JBE-treated macrophages were incubated with yeast cells pre-treated with GlcNAc, though NO release was decreased. Yeast cells were treated with mice anti-JBE IgG F(ab) polyclonal fragments (50 e 100 pg/mL) and macrophages treated with 50 mM monosaccharides promoted phagocytosis inhibition, the incubation of both promoted a increase on phagocytosis inhibition. The incubation of JBE-treated macrophages (50 pg/mL) and yeast cells pre-treated with anti-JBE IgG F(ab) (50 pg/mL) promoted phagocytosis inhibition (61,5 %). This results suggest that JBE could be involved on the fungus and host interaction because its localization. Furthermore, JBE potentializes the phagocytosis of P. brasiliensis yeast cells by murine peritoneal macrophages. Probably, the phagocytosis inhibition and NO decrease by GlcNAc is due to gp 70 affinity to GlcNAc. The anti-JBE IgG F(ab) probably occupies sites of interaction on yeast cells and improve the phagocytosis inhibition by monosaccharides or JBE on macrophages. In conclusion, JBE has component able to promote invasion of fungus.Dissertação (Mestrado)Componentes antigênicos presentes na superfície de Paracoccidioides brasiliensis são capazes de promover interação com as células do hospedeiro, favorecendo a patogênese da infecção. O presente estudo buscou verificar a presença de JBE (“Jacalin Binding Exoantigen” - Exoantígeno Ligante de Jacalina) no sobrenadante de cultura líquida de leveduras do isolado BAT de P. brasiliensis e, também, nas leveduras, bem como investigar seu efeito na fagocitose de leveduras deste fungo por macrófagos peritoneais murinos. JBE foi obtido no sobrenadante de cultura líquida ao 16° dia de cultivo de leveduras, em meio F-10 acrescido de 0,5 % de D-Glicose, sua análise, por SDS-PAGE c coloração pela prata, evidenciou uma banda de 190 kDa e, sob condições redutoras uma de 70 kDa. O ensaio imunohistoquímico utilizando IgG de coelho anti-gpl90 e IgG ligada ao ouro coloidal, localizou o componente de JBE, predominantemente, na superfície das leveduras. Macrófagos peritoneais de camundongos BALB/c foram tratados com JBE (50 e 100 pg/mL) e incubados com leveduras de P. brasiliensis marcadas com FITC pré-tratadas com 50 mM de monossacarídeos: N-acetil-D-glicosamina (GlcNAc), ou D-Glicose, ou D-Galactose ou D-Manose. JBE, quando presente nos macrófagos, foi capaz de promover a inibição da fagocitose (40 %) e a produção de NO, e ao serem incubados com leveduras pré-tratadas com GlcNAc a inibição foi ainda maior (64 %), porém houve inibição na produção de NO. As leveduras de P. brasiliensis marcadas com FITC foram tratadas com fragmentos F(ab) da IgG de camundongo anti-JBE (50 e 100 pg/mL) e os macrófagos foram tratados com 50 mM de monossacarídeos, esse tratamento promoveu inibição na fagocitose e essa inibição aumentou quando houve a incubação de ambos. A inibição da fagocitose foi verificada também em macrófagos tratados com JBE (50 pg/mL) incubados com leveduras pré-tratadas com fragmentos F(ab) da IgG de camundongo anti-JBE (50 pg/mL) (61,5 %). Esses resultados sugerem que JBE esteja envolvido na interação entre o fungo e as células do hospedeiro, inferido por sua localização, bem como, JBE mostrou-se capaz de potencializar a fagocitose de leveduras de P. brasiliensis por macrófagos peritoneais murinos. Provavelmente, a inibição da fagocitose e da produção de NO promovida por GlcNAc possa ser pela afinidade da gp70 de JBE por esse monossacarídeo. Os fragmentos F(ab) da IgG de camundongo anti-JBE, provavelmente, ocupam os sítios de interação nas leveduras e aumentam a taxa de inibição da fagocitose pelos monossacarídeos ou por JBE, nos macrófagos. Em conclusão, JBE possui componente capaz de promover a invasão do fungo.Universidade Federal de UberlândiaBrasilPrograma de Pós-graduação em Imunologia e Parasitologia AplicadasCardoso, Margareth Leitão Gennarihttp://lattes.cnpq.br/0852473414625759Carvalho, Fernanda Caroline de2019-10-11T13:30:23Z2019-10-11T13:30:23Z2004info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfCARVALHO, Fernanda Caroline de. Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos. 2004. 92 f. Dissertação (Mestrado em Imunologia e Parasitologia Aplicadas) - Universidade Federal de Uberlândia, Uberlândia, 2019. DOI http://dx.doi.org/10.14393/ufu.di.2004.28https://repositorio.ufu.br/handle/123456789/27142http://dx.doi.org/10.14393/ufu.di.2004.28porAttribution-NonCommercial-NoDerivs 3.0 United Stateshttp://creativecommons.org/licenses/by-nc-nd/3.0/us/info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFU2019-10-12T06:09:27Zoai:repositorio.ufu.br:123456789/27142Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2019-10-12T06:09:27Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.none.fl_str_mv |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos Evaluation of JBE (“Jacalin Binding Exoantigen”) in phagocytosis of Paracoccidioides brasiliensis yeast by murine peritoneal macrophages |
title |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos |
spellingShingle |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos Carvalho, Fernanda Caroline de Paracoccidioides brasiliensis Macrófagos Exoantígeno Carboidrato Paracoccidioides brasiliensis Macrophages Exoantigen Carbohydrates CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
title_short |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos |
title_full |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos |
title_fullStr |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos |
title_full_unstemmed |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos |
title_sort |
Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos |
author |
Carvalho, Fernanda Caroline de |
author_facet |
Carvalho, Fernanda Caroline de |
author_role |
author |
dc.contributor.none.fl_str_mv |
Cardoso, Margareth Leitão Gennari http://lattes.cnpq.br/0852473414625759 |
dc.contributor.author.fl_str_mv |
Carvalho, Fernanda Caroline de |
dc.subject.por.fl_str_mv |
Paracoccidioides brasiliensis Macrófagos Exoantígeno Carboidrato Paracoccidioides brasiliensis Macrophages Exoantigen Carbohydrates CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
topic |
Paracoccidioides brasiliensis Macrófagos Exoantígeno Carboidrato Paracoccidioides brasiliensis Macrophages Exoantigen Carbohydrates CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA |
description |
Paracoccidioides brasiliensis has antigenic components on surface able to promote the interaction between fungus and host, and promote the infection. In the present study the presence of JBE (Jacalin Binding Exoantigen) in culture supematant of BAT P. brasiliensis isolate and on yeast cells were verified and the effect of JBE in attachment to murine peritoneal macrophages was investigated. JBE was obtained in yeast cells culture supematant (liquid F-10 médium containing 0.5 % D-Glucose) and showed by SDS-PAGE and silver nitrate staining a 190 kDa component and, under reducing conditions, a 70 kDa component. Moreover, JBE was localized by rabbit anti-gpl90 IgG and immunogold reaction on yeast cells surface. Peritoneal macrophages form BALB/c mice were treated with JBE (50 e 100 pg/mL) and incubated with P. brasiliensis yeast cells FITC labelling pre-treated with 50 mM monosaccharides: N-acetyl-D-glucosamine (GlcNAc), or D-Glucose, or D-Galactose or D-Mannose. JBE was able to promote phagocytosis inhibition (40 %) and NO release by JBE-treated macrophages. The phagocytosis inhibition increased to 64 % when JBE-treated macrophages were incubated with yeast cells pre-treated with GlcNAc, though NO release was decreased. Yeast cells were treated with mice anti-JBE IgG F(ab) polyclonal fragments (50 e 100 pg/mL) and macrophages treated with 50 mM monosaccharides promoted phagocytosis inhibition, the incubation of both promoted a increase on phagocytosis inhibition. The incubation of JBE-treated macrophages (50 pg/mL) and yeast cells pre-treated with anti-JBE IgG F(ab) (50 pg/mL) promoted phagocytosis inhibition (61,5 %). This results suggest that JBE could be involved on the fungus and host interaction because its localization. Furthermore, JBE potentializes the phagocytosis of P. brasiliensis yeast cells by murine peritoneal macrophages. Probably, the phagocytosis inhibition and NO decrease by GlcNAc is due to gp 70 affinity to GlcNAc. The anti-JBE IgG F(ab) probably occupies sites of interaction on yeast cells and improve the phagocytosis inhibition by monosaccharides or JBE on macrophages. In conclusion, JBE has component able to promote invasion of fungus. |
publishDate |
2004 |
dc.date.none.fl_str_mv |
2004 2019-10-11T13:30:23Z 2019-10-11T13:30:23Z |
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.uri.fl_str_mv |
CARVALHO, Fernanda Caroline de. Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos. 2004. 92 f. Dissertação (Mestrado em Imunologia e Parasitologia Aplicadas) - Universidade Federal de Uberlândia, Uberlândia, 2019. DOI http://dx.doi.org/10.14393/ufu.di.2004.28 https://repositorio.ufu.br/handle/123456789/27142 http://dx.doi.org/10.14393/ufu.di.2004.28 |
identifier_str_mv |
CARVALHO, Fernanda Caroline de. Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos. 2004. 92 f. Dissertação (Mestrado em Imunologia e Parasitologia Aplicadas) - Universidade Federal de Uberlândia, Uberlândia, 2019. DOI http://dx.doi.org/10.14393/ufu.di.2004.28 |
url |
https://repositorio.ufu.br/handle/123456789/27142 http://dx.doi.org/10.14393/ufu.di.2004.28 |
dc.language.iso.fl_str_mv |
por |
language |
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Attribution-NonCommercial-NoDerivs 3.0 United States http://creativecommons.org/licenses/by-nc-nd/3.0/us/ info:eu-repo/semantics/openAccess |
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Attribution-NonCommercial-NoDerivs 3.0 United States http://creativecommons.org/licenses/by-nc-nd/3.0/us/ |
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openAccess |
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application/pdf |
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Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas |
publisher.none.fl_str_mv |
Universidade Federal de Uberlândia Brasil Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas |
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reponame:Repositório Institucional da UFU instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
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Universidade Federal de Uberlândia (UFU) |
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UFU |
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Repositório Institucional da UFU |
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Repositório Institucional da UFU |
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Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU) |
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diinf@dirbi.ufu.br |
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