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
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/83049
Ver los metadatos del registro completo
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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 |
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http://sedici.unlp.edu.ar/handle/10915/83049 |
dc.language.none.fl_str_mv |
eng |
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eng |
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info:eu-repo/semantics/altIdentifier/issn/0969-7128 info:eu-repo/semantics/altIdentifier/doi/10.1038/sj.gt.3302775 |
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info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/4.0/ Creative Commons Attribution 4.0 International (CC BY 4.0) |
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openAccess |
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http://creativecommons.org/licenses/by/4.0/ Creative Commons Attribution 4.0 International (CC BY 4.0) |
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