Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar
Autor(a) principal: | |
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Data de Publicação: | 2010 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFU |
Texto Completo: | https://repositorio.ufu.br/handle/123456789/16587 |
Resumo: | Background: Blomia tropicalis (Bt) is a common house dust mite in tropical regions and important source of allergens for the induction of respiratory allergic diseases. Most Bt allergens identified so far, however, have shown sequence homology to other dust mite allergens, such as Dermatophagoides spp. The aim of this study was to investigate major IgE-binding isoforms for screening Bt allergens. Methods: We used two-dimensional (2D)-gel electrophoresis of crude Bt extract and sera from mite-sensitized patients with positive skin prick test to Bt that were screened for specific IgE antibodies in ELISA, followed by one-dimensional (1D-) and 2D-immunoblots. Inhibition 1D-immunoblot was performed to visualize the crossreactivity with Dermatophagoides spp. From 2D gels, corresponding IgE-binding spots were excised and identified by mass spectrometry. Results: IgE-binding Bt allergens with low molecular weight (LMW, 14-16 and 25 kDa) and high molecular weight (HMW, 54-134 kDa) showed >50% recognition and were considered immunodominant bands. A high homologous inhibition rate (>80%) was demonstrated with all immunodominant bands. Nearly all HMW immunodominant components showed an expressive rate of heterologous inhibition. However, the low heterologous inhibition was found to LMW immunodominant compounds (25 and 14 kDa). More than 50 spots were identified by stained 2D gel of Bt extract, from which 15 allergens showed IgE reactivity in 2D-immunoblot. Blo t 3 isoforms with enzymatic molecular function of trypsin were identified by mass spectrometry. Blo t 11 was considered as a hypothetical allergen. Other IgE-binding isoforms, such as the enzymes aldehyde dehydrogenase (Blo t ALDH), hexosaminidase (Blo t HEXM), and unknown function (Blo t 2) were also identified. Conclusions: The results indicate several isoforms (Blo t 11, Blo t 3, Blo t 2 and probably Blo t ALDH and Blo t HEXM ) from B. tropicalis recognized by IgE antibodies in Brazilian mite-allergic patients as potential candidates for diagnosis and allergen-specific immunotherapy of allergic diseases. |
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2016-06-22T18:46:21Z2013-07-152016-06-22T18:46:21Z2010-02-22ALMEIDA, Karine Cristine de. Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar. 2010. 109 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2010.https://repositorio.ufu.br/handle/123456789/16587Background: Blomia tropicalis (Bt) is a common house dust mite in tropical regions and important source of allergens for the induction of respiratory allergic diseases. Most Bt allergens identified so far, however, have shown sequence homology to other dust mite allergens, such as Dermatophagoides spp. The aim of this study was to investigate major IgE-binding isoforms for screening Bt allergens. Methods: We used two-dimensional (2D)-gel electrophoresis of crude Bt extract and sera from mite-sensitized patients with positive skin prick test to Bt that were screened for specific IgE antibodies in ELISA, followed by one-dimensional (1D-) and 2D-immunoblots. Inhibition 1D-immunoblot was performed to visualize the crossreactivity with Dermatophagoides spp. From 2D gels, corresponding IgE-binding spots were excised and identified by mass spectrometry. Results: IgE-binding Bt allergens with low molecular weight (LMW, 14-16 and 25 kDa) and high molecular weight (HMW, 54-134 kDa) showed >50% recognition and were considered immunodominant bands. A high homologous inhibition rate (>80%) was demonstrated with all immunodominant bands. Nearly all HMW immunodominant components showed an expressive rate of heterologous inhibition. However, the low heterologous inhibition was found to LMW immunodominant compounds (25 and 14 kDa). More than 50 spots were identified by stained 2D gel of Bt extract, from which 15 allergens showed IgE reactivity in 2D-immunoblot. Blo t 3 isoforms with enzymatic molecular function of trypsin were identified by mass spectrometry. Blo t 11 was considered as a hypothetical allergen. Other IgE-binding isoforms, such as the enzymes aldehyde dehydrogenase (Blo t ALDH), hexosaminidase (Blo t HEXM), and unknown function (Blo t 2) were also identified. Conclusions: The results indicate several isoforms (Blo t 11, Blo t 3, Blo t 2 and probably Blo t ALDH and Blo t HEXM ) from B. tropicalis recognized by IgE antibodies in Brazilian mite-allergic patients as potential candidates for diagnosis and allergen-specific immunotherapy of allergic diseases.Blomia tropicalis (Bt) é um ácaro da poeira domiciliar comum em regiões tropicais e importante fonte de alérgenos para a indução de doenças alérgicas respiratórias. Até o momento, a maioria dos alérgenos de Bt identificados mostrou homologia de seqüência com outros alérgenos de ácaros da poeira, como Dermatophagoides spp. O objetivo deste estudo foi investigar os principais alérgenos com reatividade a anticorpos IgE no soro de pacientes alérgicos. Métodos: O extrato Bt foi submetido à eletroforese bidimensional (2D) e soros de pacientes sensibilizados a ácaros com teste cutâneo positivo a Bt, foram selecionados por ELISA para a presença de anticorpos IgE específicos e utilizados em ensaios de immunoblot uni- e bidimensional. Immunoblot de inibição 1D foi realizado para avaliar a reatividade cruzada com Dermatophagoides spp. Spots visualizados no gel 2D que apresentaram reatividade a IgE do soro de pacientes alérgicos foram excisados e identificados por espectrometria de massa. Resultados: Alérgenos de Bt com baixo (14-16 e 25 kDa) e alto peso molecular (54-134 kDa) apresentaram mais de 50% de reatividade a anticorpos IgE no soro de pacientes alérgicos e foram considerados imunodominantes. Uma alta taxa de inibição homóloga (> 80%) foi observada para todas as bandas imunodominantes. Quase todos os componentes imunodominantes de alto peso molecular mostraram uma taxa expressiva de inibição heteróloga. No entanto, a menor inibição heteróloga foi encontrada para os compostos imunodominantes de baixo peso molecular (25 e 14 kDa). Mais de 50 spots foram identificados pelo gel 2D, dos quais 15 alérgenos apresentaram reatividade IgE no immunoblot 2D. Isoformas do alérgeno Blo t 3, já conhecido com a função molecular enzimática da tripsina, foram identificadas por espectrometria de massa. Blo t 11 foi considerado um alérgeno hipotético. Outras isoformas ligantes de IgE, tais como as enzimas aldeído desidrogenase (Blo t ALDH) e hexosaminidase (Blot HEXM), e uma isoforma com função desconhecida (Blo t 2), também foram identificadas. Conclusões: Os alérgenos Blo t 11, Blo t 3, Blo t 2 e possivelmente Blo t ALDH e Blo t HEXM demonstraram ser alérgenos imunodominantes em pacientes sensibilizados a ácaros da poeira domiciliar de nossa região.Doutor em Imunologia e Parasitologia Aplicadasapplication/pdfporUniversidade Federal de UberlândiaPrograma de Pós-graduação em Imunologia e Parasitologia AplicadasUFUBRCiências BiológicasRinite alérgicaBlomia tropicalisReactividade cruzadaÁcaros da poeira domiciliarIgEFebre do fenoAlergiaAllergic rhinitisCross-reactivityHouse dust mite allergensCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADAIdentificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliarinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisSilva, Deise Aparecida de Oliveirahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701157D0Taketomi, Ernesto Akiohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788625D6Silva Segundo, Gesmar Rodrigueshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4777060P8Mendes, Nelson FigueiredoEspindola, Foued Salmenhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727087Y6Silva, Cláudio Vieira dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4704340J2http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4775901A4Almeida, Karine Cristine deinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFUTHUMBNAILIdentificacaoIsoformasBlomia.pdf.jpgIdentificacaoIsoformasBlomia.pdf.jpgGenerated Thumbnailimage/jpeg1426https://repositorio.ufu.br/bitstream/123456789/16587/3/IdentificacaoIsoformasBlomia.pdf.jpge96d8d538710c39f377a149c77623902MD53ORIGINALIdentificacaoIsoformasBlomia.pdfapplication/pdf3280607https://repositorio.ufu.br/bitstream/123456789/16587/1/IdentificacaoIsoformasBlomia.pdf1f6ad92973d94c110ee692cffb61f782MD51TEXTIdentificacaoIsoformasBlomia.pdf.txtIdentificacaoIsoformasBlomia.pdf.txtExtracted texttext/plain228560https://repositorio.ufu.br/bitstream/123456789/16587/2/IdentificacaoIsoformasBlomia.pdf.txt4ade1951deaf97bc5dfc5e96eeb23e70MD52123456789/165872016-06-23 04:36:35.737oai:repositorio.ufu.br:123456789/16587Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2016-06-23T07:36:35Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false |
dc.title.por.fl_str_mv |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
title |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
spellingShingle |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar Almeida, Karine Cristine de Rinite alérgica Blomia tropicalis Reactividade cruzada Ácaros da poeira domiciliar IgE Febre do feno Alergia Allergic rhinitis Cross-reactivity House dust mite allergens CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
title_short |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
title_full |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
title_fullStr |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
title_full_unstemmed |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
title_sort |
Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar |
author |
Almeida, Karine Cristine de |
author_facet |
Almeida, Karine Cristine de |
author_role |
author |
dc.contributor.advisor-co1.fl_str_mv |
Silva, Deise Aparecida de Oliveira |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701157D0 |
dc.contributor.advisor1.fl_str_mv |
Taketomi, Ernesto Akio |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788625D6 |
dc.contributor.referee1.fl_str_mv |
Silva Segundo, Gesmar Rodrigues |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4777060P8 |
dc.contributor.referee2.fl_str_mv |
Mendes, Nelson Figueiredo |
dc.contributor.referee3.fl_str_mv |
Espindola, Foued Salmen |
dc.contributor.referee3Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4727087Y6 |
dc.contributor.referee4.fl_str_mv |
Silva, Cláudio Vieira da |
dc.contributor.referee4Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4704340J2 |
dc.contributor.authorLattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4775901A4 |
dc.contributor.author.fl_str_mv |
Almeida, Karine Cristine de |
contributor_str_mv |
Silva, Deise Aparecida de Oliveira Taketomi, Ernesto Akio Silva Segundo, Gesmar Rodrigues Mendes, Nelson Figueiredo Espindola, Foued Salmen Silva, Cláudio Vieira da |
dc.subject.por.fl_str_mv |
Rinite alérgica Blomia tropicalis Reactividade cruzada Ácaros da poeira domiciliar IgE Febre do feno Alergia |
topic |
Rinite alérgica Blomia tropicalis Reactividade cruzada Ácaros da poeira domiciliar IgE Febre do feno Alergia Allergic rhinitis Cross-reactivity House dust mite allergens CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
dc.subject.eng.fl_str_mv |
Allergic rhinitis Cross-reactivity House dust mite allergens |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA |
description |
Background: Blomia tropicalis (Bt) is a common house dust mite in tropical regions and important source of allergens for the induction of respiratory allergic diseases. Most Bt allergens identified so far, however, have shown sequence homology to other dust mite allergens, such as Dermatophagoides spp. The aim of this study was to investigate major IgE-binding isoforms for screening Bt allergens. Methods: We used two-dimensional (2D)-gel electrophoresis of crude Bt extract and sera from mite-sensitized patients with positive skin prick test to Bt that were screened for specific IgE antibodies in ELISA, followed by one-dimensional (1D-) and 2D-immunoblots. Inhibition 1D-immunoblot was performed to visualize the crossreactivity with Dermatophagoides spp. From 2D gels, corresponding IgE-binding spots were excised and identified by mass spectrometry. Results: IgE-binding Bt allergens with low molecular weight (LMW, 14-16 and 25 kDa) and high molecular weight (HMW, 54-134 kDa) showed >50% recognition and were considered immunodominant bands. A high homologous inhibition rate (>80%) was demonstrated with all immunodominant bands. Nearly all HMW immunodominant components showed an expressive rate of heterologous inhibition. However, the low heterologous inhibition was found to LMW immunodominant compounds (25 and 14 kDa). More than 50 spots were identified by stained 2D gel of Bt extract, from which 15 allergens showed IgE reactivity in 2D-immunoblot. Blo t 3 isoforms with enzymatic molecular function of trypsin were identified by mass spectrometry. Blo t 11 was considered as a hypothetical allergen. Other IgE-binding isoforms, such as the enzymes aldehyde dehydrogenase (Blo t ALDH), hexosaminidase (Blo t HEXM), and unknown function (Blo t 2) were also identified. Conclusions: The results indicate several isoforms (Blo t 11, Blo t 3, Blo t 2 and probably Blo t ALDH and Blo t HEXM ) from B. tropicalis recognized by IgE antibodies in Brazilian mite-allergic patients as potential candidates for diagnosis and allergen-specific immunotherapy of allergic diseases. |
publishDate |
2010 |
dc.date.issued.fl_str_mv |
2010-02-22 |
dc.date.available.fl_str_mv |
2013-07-15 2016-06-22T18:46:21Z |
dc.date.accessioned.fl_str_mv |
2016-06-22T18:46:21Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
ALMEIDA, Karine Cristine de. Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar. 2010. 109 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2010. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufu.br/handle/123456789/16587 |
identifier_str_mv |
ALMEIDA, Karine Cristine de. Identificação de isoformas de Blomia tropicalis (Acari: Echimyopodidae) ligantes de anticorpos IgE de pacientes sensibilizados a ácaros da poeira domiciliar. 2010. 109 f. Tese (Doutorado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2010. |
url |
https://repositorio.ufu.br/handle/123456789/16587 |
dc.language.iso.fl_str_mv |
por |
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por |
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info:eu-repo/semantics/openAccess |
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openAccess |
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dc.publisher.none.fl_str_mv |
Universidade Federal de Uberlândia |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Imunologia e Parasitologia Aplicadas |
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UFU |
dc.publisher.country.fl_str_mv |
BR |
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Ciências Biológicas |
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Universidade Federal de Uberlândia |
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reponame:Repositório Institucional da UFU instname:Universidade Federal de Uberlândia (UFU) instacron:UFU |
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