A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning

Autores
Manful, Theresa; Mulindwa, Julius; Frank, Fernanda Maria; Clayton, Christine E.; Matovu, Enock
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. Methodology and Principal Findings: To find trypanosome proteins that are specifically recognised by sera from human sleeping sickness patients, we have screened the Trypanosoma brucei brucei proteome by Western blotting. Using cytosolic, cytoskeletal and glycosomal fractions, we found that the vast majority of abundant trypanosome proteins is not specifically recognised by patient sera. We identified phosphoglycerate kinase (PGKC), heat shock protein (HSP70), and histones H2B and H3 as possible candidate diagnostic antigens. These proteins, plus paraflagellar rod protein 1, rhodesain (a cysteine protease), and an extracellular fragment of the Trypanosoma brucei nucleoside transporter TbNT10, were expressed in E. coli and tested for reactivity with patient and control sera. Only TbHSP70 was preferentially recognized by patient sera, but the sensitivity and specificity were insufficient for use of TbHSP70 alone as a diagnostic. Immunoprecipitation using a native protein extract revealed no specifically reacting proteins. Conclusions: No abundant T. brucei soluble, glycosomal or cytoskeletal protein is likely to be useful in diagnosis. To find useful diagnostic antigens it will therefore be necessary to use more sophisticated proteomic methods, or to test a very large panel of candidate proteins.
Fil: Manful, Theresa. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania
Fil: Mulindwa, Julius. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania. Makerere University. Faculty of Veterinary Medicine; Uganda
Fil: Frank, Fernanda Maria. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "Profesor R. A. Margni"; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina
Fil: Clayton, Christine E.. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania
Fil: Matovu, Enock. Makerere University. Faculty of Veterinary Medicine; Uganda
Materia
Trypanosoma brucei rhodesiense
diagnosis
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
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Acceder
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloningManful, TheresaMulindwa, JuliusFrank, Fernanda MariaClayton, Christine E.Matovu, EnockTrypanosoma brucei rhodesiensediagnosishttps://purl.org/becyt/ford/3.1https://purl.org/becyt/ford/3Background: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. Methodology and Principal Findings: To find trypanosome proteins that are specifically recognised by sera from human sleeping sickness patients, we have screened the Trypanosoma brucei brucei proteome by Western blotting. Using cytosolic, cytoskeletal and glycosomal fractions, we found that the vast majority of abundant trypanosome proteins is not specifically recognised by patient sera. We identified phosphoglycerate kinase (PGKC), heat shock protein (HSP70), and histones H2B and H3 as possible candidate diagnostic antigens. These proteins, plus paraflagellar rod protein 1, rhodesain (a cysteine protease), and an extracellular fragment of the Trypanosoma brucei nucleoside transporter TbNT10, were expressed in E. coli and tested for reactivity with patient and control sera. Only TbHSP70 was preferentially recognized by patient sera, but the sensitivity and specificity were insufficient for use of TbHSP70 alone as a diagnostic. Immunoprecipitation using a native protein extract revealed no specifically reacting proteins. Conclusions: No abundant T. brucei soluble, glycosomal or cytoskeletal protein is likely to be useful in diagnosis. To find useful diagnostic antigens it will therefore be necessary to use more sophisticated proteomic methods, or to test a very large panel of candidate proteins.Fil: Manful, Theresa. Universität Heidelberg. Zentrum für Molekulare Biologie; AlemaniaFil: Mulindwa, Julius. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania. Makerere University. Faculty of Veterinary Medicine; UgandaFil: Frank, Fernanda Maria. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "Profesor R. A. Margni"; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; ArgentinaFil: Clayton, Christine E.. Universität Heidelberg. Zentrum für Molekulare Biologie; AlemaniaFil: Matovu, Enock. Makerere University. Faculty of Veterinary Medicine; UgandaPublic Library of Science2010-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/13892Manful, Theresa; Mulindwa, Julius; Frank, Fernanda Maria; Clayton, Christine E.; Matovu, Enock; A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning; Public Library of Science; Plos One; 5; e963; 3-2010; 1-71932-6203enginfo:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0009630info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0009630info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2835760/info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T12:04:09Zoai:ri.conicet.gov.ar:11336/13892instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 12:04:09.647CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
title A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
spellingShingle A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
Manful, Theresa
Trypanosoma brucei rhodesiense
diagnosis
title_short A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
title_full A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
title_fullStr A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
title_full_unstemmed A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
title_sort A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning
dc.creator.none.fl_str_mv Manful, Theresa
Mulindwa, Julius
Frank, Fernanda Maria
Clayton, Christine E.
Matovu, Enock
author Manful, Theresa
author_facet Manful, Theresa
Mulindwa, Julius
Frank, Fernanda Maria
Clayton, Christine E.
Matovu, Enock
author_role author
author2 Mulindwa, Julius
Frank, Fernanda Maria
Clayton, Christine E.
Matovu, Enock
author2_role author
author
author
author
dc.subject.none.fl_str_mv Trypanosoma brucei rhodesiense
diagnosis
topic Trypanosoma brucei rhodesiense
diagnosis
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.1
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Background: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. Methodology and Principal Findings: To find trypanosome proteins that are specifically recognised by sera from human sleeping sickness patients, we have screened the Trypanosoma brucei brucei proteome by Western blotting. Using cytosolic, cytoskeletal and glycosomal fractions, we found that the vast majority of abundant trypanosome proteins is not specifically recognised by patient sera. We identified phosphoglycerate kinase (PGKC), heat shock protein (HSP70), and histones H2B and H3 as possible candidate diagnostic antigens. These proteins, plus paraflagellar rod protein 1, rhodesain (a cysteine protease), and an extracellular fragment of the Trypanosoma brucei nucleoside transporter TbNT10, were expressed in E. coli and tested for reactivity with patient and control sera. Only TbHSP70 was preferentially recognized by patient sera, but the sensitivity and specificity were insufficient for use of TbHSP70 alone as a diagnostic. Immunoprecipitation using a native protein extract revealed no specifically reacting proteins. Conclusions: No abundant T. brucei soluble, glycosomal or cytoskeletal protein is likely to be useful in diagnosis. To find useful diagnostic antigens it will therefore be necessary to use more sophisticated proteomic methods, or to test a very large panel of candidate proteins.
Fil: Manful, Theresa. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania
Fil: Mulindwa, Julius. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania. Makerere University. Faculty of Veterinary Medicine; Uganda
Fil: Frank, Fernanda Maria. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Oficina de Coordinacion Administrativa Houssay. Instituto de Estudios de la Inmunidad Humoral "Profesor R. A. Margni"; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica; Argentina
Fil: Clayton, Christine E.. Universität Heidelberg. Zentrum für Molekulare Biologie; Alemania
Fil: Matovu, Enock. Makerere University. Faculty of Veterinary Medicine; Uganda
description Background: The only available diagnostic method for East African trypanosomiasis is light microscopy of blood samples. A simple immunodiagnostic would greatly aid trypanosomiasis control. Methodology and Principal Findings: To find trypanosome proteins that are specifically recognised by sera from human sleeping sickness patients, we have screened the Trypanosoma brucei brucei proteome by Western blotting. Using cytosolic, cytoskeletal and glycosomal fractions, we found that the vast majority of abundant trypanosome proteins is not specifically recognised by patient sera. We identified phosphoglycerate kinase (PGKC), heat shock protein (HSP70), and histones H2B and H3 as possible candidate diagnostic antigens. These proteins, plus paraflagellar rod protein 1, rhodesain (a cysteine protease), and an extracellular fragment of the Trypanosoma brucei nucleoside transporter TbNT10, were expressed in E. coli and tested for reactivity with patient and control sera. Only TbHSP70 was preferentially recognized by patient sera, but the sensitivity and specificity were insufficient for use of TbHSP70 alone as a diagnostic. Immunoprecipitation using a native protein extract revealed no specifically reacting proteins. Conclusions: No abundant T. brucei soluble, glycosomal or cytoskeletal protein is likely to be useful in diagnosis. To find useful diagnostic antigens it will therefore be necessary to use more sophisticated proteomic methods, or to test a very large panel of candidate proteins.
publishDate 2010
dc.date.none.fl_str_mv 2010-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/13892
Manful, Theresa; Mulindwa, Julius; Frank, Fernanda Maria; Clayton, Christine E.; Matovu, Enock; A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning; Public Library of Science; Plos One; 5; e963; 3-2010; 1-7
1932-6203
url http://hdl.handle.net/11336/13892
identifier_str_mv Manful, Theresa; Mulindwa, Julius; Frank, Fernanda Maria; Clayton, Christine E.; Matovu, Enock; A search for Trypanosoma brucei rhodesiense diagnostic antigens by proteomic screening and targeted cloning; Public Library of Science; Plos One; 5; e963; 3-2010; 1-7
1932-6203
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0009630
info:eu-repo/semantics/altIdentifier/doi/10.1371/journal.pone.0009630
info:eu-repo/semantics/altIdentifier/url/https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2835760/
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 12.982451

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