Avaliação de JBE (“Jacalin Binding Exoantigen”- Exoantígeno Ligante de Jacalina) na fagocitose de leveduras de Paracoccidioides brasiliensis por macrófagos peritoneais murinos

Detalhes bibliográficos
Autor(a) principal: Carvalho, Fernanda Caroline de
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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spelling 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 por
dc.rights.driver.fl_str_mv Attribution-NonCommercial-NoDerivs 3.0 United States
http://creativecommons.org/licenses/by-nc-nd/3.0/us/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Attribution-NonCommercial-NoDerivs 3.0 United States
http://creativecommons.org/licenses/by-nc-nd/3.0/us/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv 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
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFU
instname:Universidade Federal de Uberlândia (UFU)
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instname_str Universidade Federal de Uberlândia (UFU)
instacron_str UFU
institution UFU
reponame_str Repositório Institucional da UFU
collection Repositório Institucional da UFU
repository.name.fl_str_mv Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)
repository.mail.fl_str_mv diinf@dirbi.ufu.br
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