Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos

Detalhes bibliográficos
Autor(a) principal: Cavalheiro, Patrícia Bernardes
Data de Publicação: 2010
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Manancial - Repositório Digital da UFSM
Texto Completo: http://repositorio.ufsm.br/handle/1/5935
Resumo: Oomycete Pythium insisiosum is the etiologic agent of pythiosis, chronic and granulomatous disease, which affects humans and animals in tropical and subtropical areas of the world. Due to ergosterol absence in the plasmatic membrane of this microorganism, treatments based on antifungal agents have been ineffective. Immunotherapy has emerged showing promising results. In this context, the improvement of the immunotherapy was the main goal of this study, whose first purpose was to characterize the immunodominant antigens from this species (P. insidosum ATCC 58637 and P. insidiosum 210-LAPEMI) through sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE). Some proteins showing 15 kDa to 120 kDa were detected, and bands were transferred to nitrocellulose membranes in order to develop the western blot tests. The second purpose was to characterize the humoral response to pythiosis in equine, in bovine and in experimental pythiosis in rabbits, considering the no treated disease, the disease cured by immunotherapy, the disease treated with immunotherapy but not cured, and the disease cured spontaneously. To encompass all these cases we have included sera from: a) horses cured by immunotherapy; b) horses with pythiosis but not responsive to immunotherapy; c) horses with pythiosis but not treated; d) bovines with pythiosis and showing spontaneous cure; e) rabbits with experimental pythiosis but not treated; and f) rabbits with experimental pythiosis treated with immunotherapy. All specimens recognized three immunodominant proteins: 74, 33 and 32 kDa. The sera from horses cured by immunotherapy (a) and bovine sera (d) recognized another immunodominant protein, 55 kDa, which was weakly positive or negative in the other groups. Thus, the 74, 33 and 32 kDa immunodominant proteins suggest a relevant function in the humoral response to pythiosis because they were recognized by horses, rabbits and bovines. In addition, the 55 kDa antigen, which is being reported here for the first time, is likely to be involved in the cure mechanisms stimulated by immunotherapy.
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spelling 2013-05-202013-05-202010-07-26CAVALHEIRO, Patrícia Bernardes. CHARACTERIZATION OF IMMUNODOMINANT ANTIGEN OF PYTHIUM INSIDIOSUM RECOGNIZED BY ANTIBODIES OF HORSES, RABBITS AND CATTLE. 2010. 86 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2010.http://repositorio.ufsm.br/handle/1/5935Oomycete Pythium insisiosum is the etiologic agent of pythiosis, chronic and granulomatous disease, which affects humans and animals in tropical and subtropical areas of the world. Due to ergosterol absence in the plasmatic membrane of this microorganism, treatments based on antifungal agents have been ineffective. Immunotherapy has emerged showing promising results. In this context, the improvement of the immunotherapy was the main goal of this study, whose first purpose was to characterize the immunodominant antigens from this species (P. insidosum ATCC 58637 and P. insidiosum 210-LAPEMI) through sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE). Some proteins showing 15 kDa to 120 kDa were detected, and bands were transferred to nitrocellulose membranes in order to develop the western blot tests. The second purpose was to characterize the humoral response to pythiosis in equine, in bovine and in experimental pythiosis in rabbits, considering the no treated disease, the disease cured by immunotherapy, the disease treated with immunotherapy but not cured, and the disease cured spontaneously. To encompass all these cases we have included sera from: a) horses cured by immunotherapy; b) horses with pythiosis but not responsive to immunotherapy; c) horses with pythiosis but not treated; d) bovines with pythiosis and showing spontaneous cure; e) rabbits with experimental pythiosis but not treated; and f) rabbits with experimental pythiosis treated with immunotherapy. All specimens recognized three immunodominant proteins: 74, 33 and 32 kDa. The sera from horses cured by immunotherapy (a) and bovine sera (d) recognized another immunodominant protein, 55 kDa, which was weakly positive or negative in the other groups. Thus, the 74, 33 and 32 kDa immunodominant proteins suggest a relevant function in the humoral response to pythiosis because they were recognized by horses, rabbits and bovines. In addition, the 55 kDa antigen, which is being reported here for the first time, is likely to be involved in the cure mechanisms stimulated by immunotherapy.O oomiceto Pythium insidiosum é o agente etiológico da pitiose, doença granulomatosa crônica que afeta animais domésticos (eqüinos, bovinos, etc) e o homem, em regiões tropicais e subtropicais de todo mundo. A ausência de ergosterol na membrana plasmática deste microorganismo torna a antifungicoterapia muito limitada, donde a imunoterapia (a base de hifas rompidas) tem emergido com resultados promissores. A identificação de antígenos imunodominantes é fundamental para a consolidação dos imunoterápicos utilizados no tratamento da pitiose. Objetivando-se o refinamento destes imunoterápicos, o presente estudo teve como proposta inicial, caracterizar os antígenos imunodominantes desta espécie (P. insidosum ATCC 58637 e P. insidiosum 210-LAPEMI) através de eletroforese em gel de poliacrilamida (SDS-PAGE). Diversas proteínas de 120 a 15 kDa foram detectadas e, a seguir, eletrotransferidas para membranas de nitrocelulose para realização de western blot. O segundo objetivo foi caracterizar a resposta humoral à doença em eqüinos, bovinos e na infecção experimental em coelhos, considerando-se a doença curada pela imunoterapia, doença tratada pela imunoterapia e não curada, doença não tratada, e na doença evidenciando cura espontânea. Para tanto, utilizou-se o soro de: a) eqüinos doentes e curados pela imunoterapia; b) eqüinos doentes e sem resposta a imunoterapia; c) eqüinos doentes e não tratados; d) bovinos doentes e com cura espontânea; e) coelhos infectados e tratados pela imunoterapia; f) coelhos infectados e não tratados. Todos os soros reconheceram três proteínas imunodominantes: 74, 33 e 32 kDa. O soro dos eqüinos doentes curados pela imunoterapia (a) e o soro dos bovinos (d) reconheceram também outra proteína imunodominante de 55 kDa, a qual, foi fracamente reconhecida ou ausente nos demais grupos. Assim, comprova-se que as proteínas de 74, 33 e 32 kDa desempenham relevante papel na resposta imune à pitiose, pois foram imunogênicas nas três espécies avaliadas. Em adição, sugere-se que a proteína de 55 kDa possa estar envolvida no mecanismo de cura promovido pela imunoterapia; ressalta-se que esta proteína é aqui pela primeira vez relatada.application/pdfporUniversidade Federal de Santa MariaPrograma de Pós-Graduação em Ciências FarmacêuticasUFSMBRAnálises Clínicas e ToxicológicasPythium insidiosumImunoterapiaProteínas imunodominantesPythium insidiosumImmunotherapyImmunodominant proteinsCNPQ::CIENCIAS DA SAUDE::FARMACIACaracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinosCharacterization of immunodominant antigen of Pythium insidiosum recognized by antibodies of horses, rabbits and cattleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisAlves, Sydney Hartzhttp://lattes.cnpq.br/0330782478769631Santurio, Janio Moraishttp://lattes.cnpq.br/6316012260769979Pereira, Daniela Isabel Brayerhttp://lattes.cnpq.br/3382450720179401http://lattes.cnpq.br/4911392125230611Cavalheiro, Patrícia Bernardes201000000000400500500300300bfe6016c-9c1e-42ca-ab1d-db5453ed090c4dd4e59f-61cf-47f3-95a2-fc2b3d1dcb1ab3be69bf-7acc-480d-86ef-841f8e956de5d09ffaf4-4202-45bc-9969-4464c5afe88ainfo:eu-repo/semantics/openAccessreponame:Manancial - Repositório Digital da UFSMinstname:Universidade Federal de Santa Maria (UFSM)instacron:UFSMORIGINALCAVALHEIRO, PATRICIA BERNARDES.pdfapplication/pdf955804http://repositorio.ufsm.br/bitstream/1/5935/1/CAVALHEIRO%2c%20PATRICIA%20BERNARDES.pdf2e0dc6ff4500667f2b45c6cdd2038896MD51TEXTCAVALHEIRO, PATRICIA BERNARDES.pdf.txtCAVALHEIRO, PATRICIA BERNARDES.pdf.txtExtracted texttext/plain152898http://repositorio.ufsm.br/bitstream/1/5935/2/CAVALHEIRO%2c%20PATRICIA%20BERNARDES.pdf.txt9fa792e5b1cbc088f6c5b2e1301b68aeMD52THUMBNAILCAVALHEIRO, PATRICIA BERNARDES.pdf.jpgCAVALHEIRO, PATRICIA BERNARDES.pdf.jpgIM Thumbnailimage/jpeg4993http://repositorio.ufsm.br/bitstream/1/5935/3/CAVALHEIRO%2c%20PATRICIA%20BERNARDES.pdf.jpg80a457d1f1a179752ed3eebedbe81138MD531/59352022-10-11 11:56:21.516oai:repositorio.ufsm.br:1/5935Repositório Institucionalhttp://repositorio.ufsm.br/PUBhttp://repositorio.ufsm.br/oai/requestopendoar:39132022-10-11T14:56:21Manancial - Repositório Digital da UFSM - Universidade Federal de Santa Maria (UFSM)false
dc.title.por.fl_str_mv Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
dc.title.alternative.eng.fl_str_mv Characterization of immunodominant antigen of Pythium insidiosum recognized by antibodies of horses, rabbits and cattle
title Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
spellingShingle Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
Cavalheiro, Patrícia Bernardes
Pythium insidiosum
Imunoterapia
Proteínas imunodominantes
Pythium insidiosum
Immunotherapy
Immunodominant proteins
CNPQ::CIENCIAS DA SAUDE::FARMACIA
title_short Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
title_full Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
title_fullStr Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
title_full_unstemmed Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
title_sort Caracterização de antígenos imunodominantes de Pythium insidiosum reconhecidos por anticorpos de eqüinos, coelhos e bovinos
author Cavalheiro, Patrícia Bernardes
author_facet Cavalheiro, Patrícia Bernardes
author_role author
dc.contributor.advisor1.fl_str_mv Alves, Sydney Hartz
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/0330782478769631
dc.contributor.referee1.fl_str_mv Santurio, Janio Morais
dc.contributor.referee1Lattes.fl_str_mv http://lattes.cnpq.br/6316012260769979
dc.contributor.referee2.fl_str_mv Pereira, Daniela Isabel Brayer
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/3382450720179401
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/4911392125230611
dc.contributor.author.fl_str_mv Cavalheiro, Patrícia Bernardes
contributor_str_mv Alves, Sydney Hartz
Santurio, Janio Morais
Pereira, Daniela Isabel Brayer
dc.subject.por.fl_str_mv Pythium insidiosum
Imunoterapia
Proteínas imunodominantes
topic Pythium insidiosum
Imunoterapia
Proteínas imunodominantes
Pythium insidiosum
Immunotherapy
Immunodominant proteins
CNPQ::CIENCIAS DA SAUDE::FARMACIA
dc.subject.eng.fl_str_mv Pythium insidiosum
Immunotherapy
Immunodominant proteins
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS DA SAUDE::FARMACIA
description Oomycete Pythium insisiosum is the etiologic agent of pythiosis, chronic and granulomatous disease, which affects humans and animals in tropical and subtropical areas of the world. Due to ergosterol absence in the plasmatic membrane of this microorganism, treatments based on antifungal agents have been ineffective. Immunotherapy has emerged showing promising results. In this context, the improvement of the immunotherapy was the main goal of this study, whose first purpose was to characterize the immunodominant antigens from this species (P. insidosum ATCC 58637 and P. insidiosum 210-LAPEMI) through sodium dodecyl sulfate polyacrylamide gel eletrophoresis (SDS-PAGE). Some proteins showing 15 kDa to 120 kDa were detected, and bands were transferred to nitrocellulose membranes in order to develop the western blot tests. The second purpose was to characterize the humoral response to pythiosis in equine, in bovine and in experimental pythiosis in rabbits, considering the no treated disease, the disease cured by immunotherapy, the disease treated with immunotherapy but not cured, and the disease cured spontaneously. To encompass all these cases we have included sera from: a) horses cured by immunotherapy; b) horses with pythiosis but not responsive to immunotherapy; c) horses with pythiosis but not treated; d) bovines with pythiosis and showing spontaneous cure; e) rabbits with experimental pythiosis but not treated; and f) rabbits with experimental pythiosis treated with immunotherapy. All specimens recognized three immunodominant proteins: 74, 33 and 32 kDa. The sera from horses cured by immunotherapy (a) and bovine sera (d) recognized another immunodominant protein, 55 kDa, which was weakly positive or negative in the other groups. Thus, the 74, 33 and 32 kDa immunodominant proteins suggest a relevant function in the humoral response to pythiosis because they were recognized by horses, rabbits and bovines. In addition, the 55 kDa antigen, which is being reported here for the first time, is likely to be involved in the cure mechanisms stimulated by immunotherapy.
publishDate 2010
dc.date.issued.fl_str_mv 2010-07-26
dc.date.accessioned.fl_str_mv 2013-05-20
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dc.identifier.citation.fl_str_mv CAVALHEIRO, Patrícia Bernardes. CHARACTERIZATION OF IMMUNODOMINANT ANTIGEN OF PYTHIUM INSIDIOSUM RECOGNIZED BY ANTIBODIES OF HORSES, RABBITS AND CATTLE. 2010. 86 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2010.
dc.identifier.uri.fl_str_mv http://repositorio.ufsm.br/handle/1/5935
identifier_str_mv CAVALHEIRO, Patrícia Bernardes. CHARACTERIZATION OF IMMUNODOMINANT ANTIGEN OF PYTHIUM INSIDIOSUM RECOGNIZED BY ANTIBODIES OF HORSES, RABBITS AND CATTLE. 2010. 86 f. Dissertação (Mestrado em Farmacologia) - Universidade Federal de Santa Maria, Santa Maria, 2010.
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