Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins
| Autor(a) principal: | |
|---|---|
| Data de Publicação: | 2014 |
| Outros Autores: | , , , , , , , , , , , , , |
| Tipo de documento: | Artigo |
| Idioma: | eng |
| Título da fonte: | Repositório Institucional da FIOCRUZ (ARCA) |
| Texto Completo: | https://www.arca.fiocruz.br/handle/icict/9802 |
Resumo: | University of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United Kingdom/University of Nottingham. School of Biosciences. Loughborough, United Kingdom |
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Wan, DanielRibeiro, Fernanda LudolfAlanine, Daniel G. W.Stretton, OwenAli, Eman AliAl-Barwary, NafalWang, XiaoweiDoenhoff, Michael J.Mari, AdrianoFitzsimmons, Colin M.Dunne, David W.Nakamura, RyosukeOliveira, Guilherme Correa deAlcocer, Marcos J. C.Falcone, Franco H.2015-03-23T13:06:28Z2015-03-23T13:06:28Z2014WAN, Daniel et al. Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins. PLoS Negl Trop Dis, v. 8, n. 9, p. e3124, 20141935-2735https://www.arca.fiocruz.br/handle/icict/980210.1371/journal.pntd.0003124engLibrary Public ScienceUse of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteinsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleUniversity of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United Kingdom/University of Nottingham. School of Biosciences. Loughborough, United KingdomFundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Grupo de Genômica e Biologia Computational. Belo Horizonte, MG, Brasil/ Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Instituto de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, BrasilUniversity of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United Kingdom/University of Nottingham. School of Biosciences. Loughborough, United KingdomUniversity of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United KingdomUniversity of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United KingdomUniversity of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United KingdomUniversity of Nottingham. School of Biosciences. Loughborough, United KingdomUniversity of Nottingham. School of Life Sciences. Nottingham, United KingdomCenter for Molecular Allergology. IDI-IRCCS. Rome, Italy/Associated Centres for Molecular Allergology. Rome, ItalyUniversity of Cambridge. Department of Pathology. Cambridge, United KingdomUniversity of Cambridge. Department of Pathology. Cambridge, United KingdomNational Institute of Health Sciences. Division of Medicinal Safety Science. Setagayaku, Tokyo, JapanFundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Grupo de Genômica e Biologia Computational. Belo Horizonte, MG, Brasil/ Fundação Oswaldo Cruz. Centro de Pesquisa René Rachou. Instituto de Ciência e Tecnologia em Doenças Tropicais. Belo Horizonte, MG, BrasilUniversity of Nottingham. School of Biosciences. Loughborough, United KingdomUniversity of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United KingdomBackground: Parasite-specific IgE is thought to correlate with protection against Schistosoma mansoni infection or re-infection. Only a few molecular targets of the IgE response in S. mansoni infection have been characterised. A better insight into the basic mechanisms of anti-parasite immunity could be gained from a genome-wide characterisation of such S. mansoni allergens. This would have repercussions on our understanding of allergy and the development of safe and efficacious vaccinations against helminthic parasites. Methodology/Principal Findings: A complete medium- to high-throughput amenable workflow, including important quality controls, is described, which enables the rapid translation of S. mansoni proteins using wheat germ lysate and subsequent assessment of potential allergenicity with a humanised Rat Basophilic Leukemia (RBL) reporter cell line. Cell-free translation is completed within 90 minutes, generating sufficient amounts of parasitic protein for rapid screening of allergenicity without any need for purification. Antigenic integrity is demonstrated using Western Blotting. After overnight incubation with infected individuals' serum, the RS-ATL8 reporter cell line is challenged with the complete wheat germ translation mixture and Luciferase activity measured, reporting cellular activation by the suspected allergen. The suitability of this system for characterization of novel S. mansoni allergens is demonstrated using well characterised plant and parasitic allergens such as Par j 2, SmTAL-1 and the IgE binding factor IPSE/alpha-1, expressed in wheat germ lysates and/or E. coli. SmTAL-1, but not SmTAL2 (used as a negative control), was able to activate the basophil reporter cell line. Conclusion/Significance: This method offers an accessible way for assessment of potential allergenicity of anti-helminthic vaccine candidates and is suitable for medium- to high-throughput studies using infected individual sera. It is also suitable for the study of the basis of allergenicity of helminthic proteins.Schistosoma mansoniallergenicityhelminthic proteinsinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82352https://www.arca.fiocruz.br/bitstream/icict/9802/1/license.txtafbdf7d7a9bcf771a1cc7edf5f618413MD51ORIGINAL2014_160.pdf2014_160.pdfapplication/pdf1390473https://www.arca.fiocruz.br/bitstream/icict/9802/2/2014_160.pdf7bbb1168830c3ca3293dc6c532318a34MD52TEXT2014_160.pdf.txt2014_160.pdf.txtExtracted texttext/plain63931https://www.arca.fiocruz.br/bitstream/icict/9802/3/2014_160.pdf.txt054ecae28051a0217bcbbf1f044fbb28MD53icict/98022019-06-19 10:12:10.184oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352019-06-19T13:12:10Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false |
| dc.title.pt_BR.fl_str_mv |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| title |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| spellingShingle |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins Wan, Daniel Schistosoma mansoni allergenicity helminthic proteins |
| title_short |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| title_full |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| title_fullStr |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| title_full_unstemmed |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| title_sort |
Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins |
| author |
Wan, Daniel |
| author_facet |
Wan, Daniel Ribeiro, Fernanda Ludolf Alanine, Daniel G. W. Stretton, Owen Ali, Eman Ali Al-Barwary, Nafal Wang, Xiaowei Doenhoff, Michael J. Mari, Adriano Fitzsimmons, Colin M. Dunne, David W. Nakamura, Ryosuke Oliveira, Guilherme Correa de Alcocer, Marcos J. C. Falcone, Franco H. |
| author_role |
author |
| author2 |
Ribeiro, Fernanda Ludolf Alanine, Daniel G. W. Stretton, Owen Ali, Eman Ali Al-Barwary, Nafal Wang, Xiaowei Doenhoff, Michael J. Mari, Adriano Fitzsimmons, Colin M. Dunne, David W. Nakamura, Ryosuke Oliveira, Guilherme Correa de Alcocer, Marcos J. C. Falcone, Franco H. |
| author2_role |
author author author author author author author author author author author author author author |
| dc.contributor.author.fl_str_mv |
Wan, Daniel Ribeiro, Fernanda Ludolf Alanine, Daniel G. W. Stretton, Owen Ali, Eman Ali Al-Barwary, Nafal Wang, Xiaowei Doenhoff, Michael J. Mari, Adriano Fitzsimmons, Colin M. Dunne, David W. Nakamura, Ryosuke Oliveira, Guilherme Correa de Alcocer, Marcos J. C. Falcone, Franco H. |
| dc.subject.en.pt_BR.fl_str_mv |
Schistosoma mansoni allergenicity helminthic proteins |
| topic |
Schistosoma mansoni allergenicity helminthic proteins |
| description |
University of Nottingham. School of Pharmacy. Division of Molecular and Cellular Science. Nottingham, United Kingdom/University of Nottingham. School of Biosciences. Loughborough, United Kingdom |
| publishDate |
2014 |
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2014 |
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2015-03-23T13:06:28Z |
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2015-03-23T13:06:28Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
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article |
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publishedVersion |
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WAN, Daniel et al. Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins. PLoS Negl Trop Dis, v. 8, n. 9, p. e3124, 2014 |
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https://www.arca.fiocruz.br/handle/icict/9802 |
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1935-2735 |
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10.1371/journal.pntd.0003124 |
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WAN, Daniel et al. Use of Humanised Rat Basophilic Leukaemia Cell Line RSATL8 for the Assessment of Allergenicity of Schistosoma mansoni Proteins. PLoS Negl Trop Dis, v. 8, n. 9, p. e3124, 2014 1935-2735 10.1371/journal.pntd.0003124 |
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Library Public Science |
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