Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats

Autores
Reggiani, Paula Cecilia; Hereñú, Claudia Beatriz; Rimoldi, Omar Jorge; Brown, Oscar Alfredo; Pléau, J. M.; Dardenne, M.; Goya, Rodolfo Gustavo
Año de publicación
2006
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Thymulin is a thymic peptide possessing hypophysiotropic activity and antiinflammatory effects in the brain. We constructed a synthetic DNA sequence encoding met-FTS, a biologically active analog of thymulin, and subsequently cloned it into different expression vectors. A sequence optimized for expression of met-FTS in rodents, 5′-ATGCAGGCCAAGTCGCAGGGGGGGTCGAACTAGTAG-3′, was cloned in the mammalian expression vectors pCDNA3.1(+) and phMGFP (which expresses the Monster Green Fluorescent Protein), thus obtaining pcDNA3.1-metFTS and p-metFTS-hMGFP, which express met-FTS and the fluorescent fusion protein metFTS-hMGFP, respectively. The synthetic sequence was also used to construct the adenoviral vector RAd-metFTS, which expresses met-FTS. Transfection of HEK293 and BHK cells with pcDNA3.1-metFTS (experimental groups) or pcDNA3.1 (control), led to high levels of thymulin bioactivity (>600 versus < 0.1 pg/ml in experimental and control supernatants, respectively). Transfection of HEK293 and BHK cells with pmetFTS-hMGFP revealed a cytoplasmic and nuclear distribution of the fluorescent fusion protein. A single intramuscular (i.m.) injection (107 plaque forming units (PFU)/mouse or 108/PFU/rat) of RAd-metFTS in thymectomized animals (nondetectable serum thymulin) restored serum thymulin levels for at least 110 and 130 days post-injection in mice and rats, respectively. We conclude that RAd-metFTS constitutes a suitable biotechnological tool for the implementation of thymulin gene therapy in animal models of chronic brain inflammation.
Facultad de Ciencias Médicas
Materia
Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/83049

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network_name_str SEDICI (UNLP)
spelling Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and ratsReggiani, Paula CeciliaHereñú, Claudia BeatrizRimoldi, Omar JorgeBrown, Oscar AlfredoPléau, J. M.Dardenne, M.Goya, Rodolfo GustavoBioquímicaThymulinFluorescent fusion proteinThymectomized animalsThymulin is a thymic peptide possessing hypophysiotropic activity and antiinflammatory effects in the brain. We constructed a synthetic DNA sequence encoding met-FTS, a biologically active analog of thymulin, and subsequently cloned it into different expression vectors. A sequence optimized for expression of met-FTS in rodents, 5′-ATGCAGGCCAAGTCGCAGGGGGGGTCGAACTAGTAG-3′, was cloned in the mammalian expression vectors pCDNA3.1(+) and phMGFP (which expresses the Monster Green Fluorescent Protein), thus obtaining pcDNA3.1-metFTS and p-metFTS-hMGFP, which express met-FTS and the fluorescent fusion protein metFTS-hMGFP, respectively. The synthetic sequence was also used to construct the adenoviral vector RAd-metFTS, which expresses met-FTS. Transfection of HEK293 and BHK cells with pcDNA3.1-metFTS (experimental groups) or pcDNA3.1 (control), led to high levels of thymulin bioactivity (>600 versus < 0.1 pg/ml in experimental and control supernatants, respectively). Transfection of HEK293 and BHK cells with pmetFTS-hMGFP revealed a cytoplasmic and nuclear distribution of the fluorescent fusion protein. A single intramuscular (i.m.) injection (107 plaque forming units (PFU)/mouse or 108/PFU/rat) of RAd-metFTS in thymectomized animals (nondetectable serum thymulin) restored serum thymulin levels for at least 110 and 130 days post-injection in mice and rats, respectively. We conclude that RAd-metFTS constitutes a suitable biotechnological tool for the implementation of thymulin gene therapy in animal models of chronic brain inflammation.Facultad de Ciencias Médicas2006-04-15info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf1214-1221http://sedici.unlp.edu.ar/handle/10915/83049enginfo:eu-repo/semantics/altIdentifier/issn/0969-7128info:eu-repo/semantics/altIdentifier/doi/10.1038/sj.gt.3302775info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by/4.0/Creative Commons Attribution 4.0 International (CC BY 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:15:45Zoai:sedici.unlp.edu.ar:10915/83049Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:15:46.049SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
title Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
spellingShingle Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
Reggiani, Paula Cecilia
Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
title_short Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
title_full Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
title_fullStr Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
title_full_unstemmed Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
title_sort Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
dc.creator.none.fl_str_mv Reggiani, Paula Cecilia
Hereñú, Claudia Beatriz
Rimoldi, Omar Jorge
Brown, Oscar Alfredo
Pléau, J. M.
Dardenne, M.
Goya, Rodolfo Gustavo
author Reggiani, Paula Cecilia
author_facet Reggiani, Paula Cecilia
Hereñú, Claudia Beatriz
Rimoldi, Omar Jorge
Brown, Oscar Alfredo
Pléau, J. M.
Dardenne, M.
Goya, Rodolfo Gustavo
author_role author
author2 Hereñú, Claudia Beatriz
Rimoldi, Omar Jorge
Brown, Oscar Alfredo
Pléau, J. M.
Dardenne, M.
Goya, Rodolfo Gustavo
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
topic Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
dc.description.none.fl_txt_mv Thymulin is a thymic peptide possessing hypophysiotropic activity and antiinflammatory effects in the brain. We constructed a synthetic DNA sequence encoding met-FTS, a biologically active analog of thymulin, and subsequently cloned it into different expression vectors. A sequence optimized for expression of met-FTS in rodents, 5′-ATGCAGGCCAAGTCGCAGGGGGGGTCGAACTAGTAG-3′, was cloned in the mammalian expression vectors pCDNA3.1(+) and phMGFP (which expresses the Monster Green Fluorescent Protein), thus obtaining pcDNA3.1-metFTS and p-metFTS-hMGFP, which express met-FTS and the fluorescent fusion protein metFTS-hMGFP, respectively. The synthetic sequence was also used to construct the adenoviral vector RAd-metFTS, which expresses met-FTS. Transfection of HEK293 and BHK cells with pcDNA3.1-metFTS (experimental groups) or pcDNA3.1 (control), led to high levels of thymulin bioactivity (>600 versus < 0.1 pg/ml in experimental and control supernatants, respectively). Transfection of HEK293 and BHK cells with pmetFTS-hMGFP revealed a cytoplasmic and nuclear distribution of the fluorescent fusion protein. A single intramuscular (i.m.) injection (107 plaque forming units (PFU)/mouse or 108/PFU/rat) of RAd-metFTS in thymectomized animals (nondetectable serum thymulin) restored serum thymulin levels for at least 110 and 130 days post-injection in mice and rats, respectively. We conclude that RAd-metFTS constitutes a suitable biotechnological tool for the implementation of thymulin gene therapy in animal models of chronic brain inflammation.
Facultad de Ciencias Médicas
description Thymulin is a thymic peptide possessing hypophysiotropic activity and antiinflammatory effects in the brain. We constructed a synthetic DNA sequence encoding met-FTS, a biologically active analog of thymulin, and subsequently cloned it into different expression vectors. A sequence optimized for expression of met-FTS in rodents, 5′-ATGCAGGCCAAGTCGCAGGGGGGGTCGAACTAGTAG-3′, was cloned in the mammalian expression vectors pCDNA3.1(+) and phMGFP (which expresses the Monster Green Fluorescent Protein), thus obtaining pcDNA3.1-metFTS and p-metFTS-hMGFP, which express met-FTS and the fluorescent fusion protein metFTS-hMGFP, respectively. The synthetic sequence was also used to construct the adenoviral vector RAd-metFTS, which expresses met-FTS. Transfection of HEK293 and BHK cells with pcDNA3.1-metFTS (experimental groups) or pcDNA3.1 (control), led to high levels of thymulin bioactivity (>600 versus < 0.1 pg/ml in experimental and control supernatants, respectively). Transfection of HEK293 and BHK cells with pmetFTS-hMGFP revealed a cytoplasmic and nuclear distribution of the fluorescent fusion protein. A single intramuscular (i.m.) injection (107 plaque forming units (PFU)/mouse or 108/PFU/rat) of RAd-metFTS in thymectomized animals (nondetectable serum thymulin) restored serum thymulin levels for at least 110 and 130 days post-injection in mice and rats, respectively. We conclude that RAd-metFTS constitutes a suitable biotechnological tool for the implementation of thymulin gene therapy in animal models of chronic brain inflammation.
publishDate 2006
dc.date.none.fl_str_mv 2006-04-15
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/83049
url http://sedici.unlp.edu.ar/handle/10915/83049
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/0969-7128
info:eu-repo/semantics/altIdentifier/doi/10.1038/sj.gt.3302775
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International (CC BY 4.0)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by/4.0/
Creative Commons Attribution 4.0 International (CC BY 4.0)
dc.format.none.fl_str_mv application/pdf
1214-1221
dc.source.none.fl_str_mv reponame:SEDICI (UNLP)
instname:Universidad Nacional de La Plata
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collection SEDICI (UNLP)
instname_str Universidad Nacional de La Plata
instacron_str UNLP
institution UNLP
repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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