Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão

Detalhes bibliográficos
Autor(a) principal: Carvalho, Edson Fernando Goulart de
Data de Publicação: 2014
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFU
Texto Completo: https://repositorio.ufu.br/handle/123456789/16708
https://doi.org/10.14393/ufu.di.2014.360
Resumo: The nematode Strongyloides stercoralis responsible for strongyloidiasis diagnosed by detection of larvae in feces. However, the elimination of low larvae hampers the detection of disease particularly in cases of immunosuppression. Immunodiagnostic tests have been developed, but the difficulty of obtaining larvae of S. stercoralis for the production of homologous antigenic extract, thereby Strongyloides venezuelensis and their different evolutionary forms for the production of antigen extracts. The aim of this study was to evaluate the detection of antibodies anti S. venezuelensis against alkaline extracts of L3 larvae, parthenogenetic females or eggs in experimental strongyloidiasis associated with immunosuppression. Male rats were divided into two groups: non-immunosuppressed and immunosuppressed experimentally infected and serum from all animals were obtained at 0, 5th, 8th, 13th and 21th days post infection (d.p.i.). The test enzyme-linked immunosorbent assay (ELISA) was performed to compare the reactive potential of alkaline extracts of L3 larvae, parthenogenetic females and eggs of S. venezuelensis in rats non-immunosuppressed and immunosuppressed. Immunoblotting was also evaluated for detection of IgG anti S. venezuelensis in both groups front to alkaline of the three developmental forms of the parasite extracts. In all comparative ELISA extracts were skilled in detecting IgG S. venezuelensis. The extract of larvae showed reactivity from the 5th dpi ceasing at 21th d.p.i in the non-immunosuppressed group while the group immunosuppressed this reactivity remained at 21th d.p.i. immunoblotting showed the reactivity profile with immunoreactive fractions in immunosuppressed group from the 5th d.p.i. while the non-immunosuppressed group reactivity from 8th d.p.i. The alkali extracts of parthenogenetic females and eggs showed a similar profile of reactivity in both ELISA and immunoblotting. Immunoreactive protein fractions of 26 and 17 kDa present in the alkaline extract of larvae presented as important markers of infection in immunosuppressed animals. It is concluded that all alkali extracts have diagnostic potential in experimental strongyloidiasis especially the larval extract in cases of immunosuppression, and immunoblotting an important complementary diagnostic tool.
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spelling 2016-06-22T18:46:42Z2014-12-052016-06-22T18:46:42Z2014-07-28CARVALHO, Edson Fernando Goulart de. Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão. 2014. 57 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2014. Disponível em: https://doi.org/10.14393/ufu.di.2014.360https://repositorio.ufu.br/handle/123456789/16708https://doi.org/10.14393/ufu.di.2014.360The nematode Strongyloides stercoralis responsible for strongyloidiasis diagnosed by detection of larvae in feces. However, the elimination of low larvae hampers the detection of disease particularly in cases of immunosuppression. Immunodiagnostic tests have been developed, but the difficulty of obtaining larvae of S. stercoralis for the production of homologous antigenic extract, thereby Strongyloides venezuelensis and their different evolutionary forms for the production of antigen extracts. The aim of this study was to evaluate the detection of antibodies anti S. venezuelensis against alkaline extracts of L3 larvae, parthenogenetic females or eggs in experimental strongyloidiasis associated with immunosuppression. Male rats were divided into two groups: non-immunosuppressed and immunosuppressed experimentally infected and serum from all animals were obtained at 0, 5th, 8th, 13th and 21th days post infection (d.p.i.). The test enzyme-linked immunosorbent assay (ELISA) was performed to compare the reactive potential of alkaline extracts of L3 larvae, parthenogenetic females and eggs of S. venezuelensis in rats non-immunosuppressed and immunosuppressed. Immunoblotting was also evaluated for detection of IgG anti S. venezuelensis in both groups front to alkaline of the three developmental forms of the parasite extracts. In all comparative ELISA extracts were skilled in detecting IgG S. venezuelensis. The extract of larvae showed reactivity from the 5th dpi ceasing at 21th d.p.i in the non-immunosuppressed group while the group immunosuppressed this reactivity remained at 21th d.p.i. immunoblotting showed the reactivity profile with immunoreactive fractions in immunosuppressed group from the 5th d.p.i. while the non-immunosuppressed group reactivity from 8th d.p.i. The alkali extracts of parthenogenetic females and eggs showed a similar profile of reactivity in both ELISA and immunoblotting. Immunoreactive protein fractions of 26 and 17 kDa present in the alkaline extract of larvae presented as important markers of infection in immunosuppressed animals. It is concluded that all alkali extracts have diagnostic potential in experimental strongyloidiasis especially the larval extract in cases of immunosuppression, and immunoblotting an important complementary diagnostic tool.O nematódeo Strongyloides stercoralis é responsável pela estrongiloidíase cujo diagnóstico é realizado pela detecção de larvas nas fezes. Porém a baixa eliminação de larvas dificulta a detecção da doença em especial em casos de imunossupressão. Testes imunodiagnósticos têm sido desenvolvidos, porém há dificuldade de se obter larvas de S. stercoralis para a produção de extrato antigênico homólogo assim o parasito Strongyloides venezuelensis e suas diferentes formas evolutivas são utilizados para produção de extratos antigênicos. O objetivo do presente estudo foi avaliar a detecção de anticorpos anti S. venezuelensis a partir de extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos na estrongiloidíase experimental associada à imunossupressão. Ratos machos foram divididos em dois grupos: não-imunossuprimidos e imunossuprimidos experimentalmente infectados e o soro de todos os animais foram obtidos no 0, 5º, 8º, 13º e 21º dias pós-infecção (d.p.i.). O teste enzyme-linked immunosorbent assay (ELISA) foi realizado para comparar o potencial reativo dos extratos alcalinos de larva L3, fêmeas partenogenéticas e ovos de S. venezuelensis em ratos não-imunossuprimidos e imunossuprimido. Immunoblotting foi avaliado quanto sua detecção de IgG anti S. venezuelensis em ambos os grupos frente aos extratos alcalinos das três formas evolutivas do parasito. No ELISA comparativo todos extratos foram efetivos na detecção de anti-IgG de S. venezuelensis. O extrato de larvas apresentou reatividade a partir do 5º d.p.i cessando no 21º d.p.i. no grupo não-imunossuprimido enquanto o grupo imunossuprimido essa reatividade permaneceu no 21º d.p.i.. O immunoblotting apresentou perfil de reatividade com frações imunorreativas no grupo imunossuprimido a partir do 5º d.p.i. enquanto o grupo não-imunossuprimido a reatividade a partir do 8º d.p.i. Os extratos alcalinos de fêmeas partenogenéticas e de ovos apresentaram similar perfil de reatividade tanto no ELISA comparativo quanto no immunoblotting. Frações proteicas imunorreativas de 26 e 17 kDa presentes no extrato alcalino de larvas apresentaram como importantes marcadores de infecção em animais imunossuprimidos. Conclui-se que todos os extratos alcalinos possuem potencial diagnóstico na estrongiloidíase experimental com destaque para o extrato de larvas em casos de imunossupressão, sendo o immunoblotting uma importante ferramenta diagnóstica complementar.Fundação de Amparo a Pesquisa do Estado de Minas GeraisMestre em Imunologia e Parasitologia Aplicadasapplication/pdfporUniversidade Federal de UberlândiaPrograma de Pós-graduação em Imunologia e Parasitologia AplicadasUFUBRCiências BiológicasImunossupressãoStrongyloides venezuelensisImmunoblottingRatosImmunosuppressionRatCNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADAImmunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressãoinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisCunha Junior, Jair Pereira dahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4795802Y5Costa-Cruz, Julia Mariahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797155D9http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4291535J3Carvalho, Edson Fernando Goulart de817615714760f38e-67c9-4f29-9dee-77c4e5a21ac4info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFUinstname:Universidade Federal de Uberlândia (UFU)instacron:UFUTHUMBNAILImmunoblottingUtilizandoExtratos.pdf.jpgImmunoblottingUtilizandoExtratos.pdf.jpgGenerated Thumbnailimage/jpeg1194https://repositorio.ufu.br/bitstream/123456789/16708/3/ImmunoblottingUtilizandoExtratos.pdf.jpg615260487eb3acc0284d0f904ef2a336MD53ORIGINALImmunoblottingUtilizandoExtratos.pdfapplication/pdf960361https://repositorio.ufu.br/bitstream/123456789/16708/1/ImmunoblottingUtilizandoExtratos.pdf0fb4723952939429a8d4d7bfd1d21c35MD51TEXTImmunoblottingUtilizandoExtratos.pdf.txtImmunoblottingUtilizandoExtratos.pdf.txtExtracted texttext/plain103244https://repositorio.ufu.br/bitstream/123456789/16708/2/ImmunoblottingUtilizandoExtratos.pdf.txt7d684cbeeda34432ff01bb3574050499MD52123456789/167082021-09-16 14:59:03.303oai:repositorio.ufu.br:123456789/16708Repositório InstitucionalONGhttp://repositorio.ufu.br/oai/requestdiinf@dirbi.ufu.bropendoar:2021-09-16T17:59:03Repositório Institucional da UFU - Universidade Federal de Uberlândia (UFU)false
dc.title.por.fl_str_mv Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
title Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
spellingShingle Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
Carvalho, Edson Fernando Goulart de
Imunossupressão
Strongyloides venezuelensis
Immunoblotting
Ratos
Immunosuppression
Rat
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA
title_short Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
title_full Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
title_fullStr Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
title_full_unstemmed Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
title_sort Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão
author Carvalho, Edson Fernando Goulart de
author_facet Carvalho, Edson Fernando Goulart de
author_role author
dc.contributor.advisor-co1.fl_str_mv Cunha Junior, Jair Pereira da
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4795802Y5
dc.contributor.advisor1.fl_str_mv Costa-Cruz, Julia Maria
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797155D9
dc.contributor.authorLattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4291535J3
dc.contributor.author.fl_str_mv Carvalho, Edson Fernando Goulart de
contributor_str_mv Cunha Junior, Jair Pereira da
Costa-Cruz, Julia Maria
dc.subject.por.fl_str_mv Imunossupressão
Strongyloides venezuelensis
Immunoblotting
Ratos
topic Imunossupressão
Strongyloides venezuelensis
Immunoblotting
Ratos
Immunosuppression
Rat
CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA
dc.subject.eng.fl_str_mv Immunosuppression
Rat
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA::IMUNOLOGIA APLICADA
description The nematode Strongyloides stercoralis responsible for strongyloidiasis diagnosed by detection of larvae in feces. However, the elimination of low larvae hampers the detection of disease particularly in cases of immunosuppression. Immunodiagnostic tests have been developed, but the difficulty of obtaining larvae of S. stercoralis for the production of homologous antigenic extract, thereby Strongyloides venezuelensis and their different evolutionary forms for the production of antigen extracts. The aim of this study was to evaluate the detection of antibodies anti S. venezuelensis against alkaline extracts of L3 larvae, parthenogenetic females or eggs in experimental strongyloidiasis associated with immunosuppression. Male rats were divided into two groups: non-immunosuppressed and immunosuppressed experimentally infected and serum from all animals were obtained at 0, 5th, 8th, 13th and 21th days post infection (d.p.i.). The test enzyme-linked immunosorbent assay (ELISA) was performed to compare the reactive potential of alkaline extracts of L3 larvae, parthenogenetic females and eggs of S. venezuelensis in rats non-immunosuppressed and immunosuppressed. Immunoblotting was also evaluated for detection of IgG anti S. venezuelensis in both groups front to alkaline of the three developmental forms of the parasite extracts. In all comparative ELISA extracts were skilled in detecting IgG S. venezuelensis. The extract of larvae showed reactivity from the 5th dpi ceasing at 21th d.p.i in the non-immunosuppressed group while the group immunosuppressed this reactivity remained at 21th d.p.i. immunoblotting showed the reactivity profile with immunoreactive fractions in immunosuppressed group from the 5th d.p.i. while the non-immunosuppressed group reactivity from 8th d.p.i. The alkali extracts of parthenogenetic females and eggs showed a similar profile of reactivity in both ELISA and immunoblotting. Immunoreactive protein fractions of 26 and 17 kDa present in the alkaline extract of larvae presented as important markers of infection in immunosuppressed animals. It is concluded that all alkali extracts have diagnostic potential in experimental strongyloidiasis especially the larval extract in cases of immunosuppression, and immunoblotting an important complementary diagnostic tool.
publishDate 2014
dc.date.available.fl_str_mv 2014-12-05
2016-06-22T18:46:42Z
dc.date.issued.fl_str_mv 2014-07-28
dc.date.accessioned.fl_str_mv 2016-06-22T18:46:42Z
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dc.identifier.citation.fl_str_mv CARVALHO, Edson Fernando Goulart de. Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão. 2014. 57 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2014. Disponível em: https://doi.org/10.14393/ufu.di.2014.360
dc.identifier.uri.fl_str_mv https://repositorio.ufu.br/handle/123456789/16708
dc.identifier.doi.por.fl_str_mv https://doi.org/10.14393/ufu.di.2014.360
identifier_str_mv CARVALHO, Edson Fernando Goulart de. Immunoblotting utilizando extratos alcalinos de larvas L3, fêmeas partenogenéticas ou ovos de Strongyloides venezuelensis na estrongiloidíase experimental associada à imunossupressão. 2014. 57 f. Dissertação (Mestrado em Ciências Biológicas) - Universidade Federal de Uberlândia, Uberlândia, 2014. Disponível em: https://doi.org/10.14393/ufu.di.2014.360
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