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

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 expr...

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Detalles Bibliográficos
Autores principales: Reggiani, Paula Cecilia, Hereñú, Claudia Beatriz, Rimoldi, Omar Jorge, Brown, Oscar Alfredo, Pléau, J. M., Dardenne, M., Goya, Rodolfo Gustavo
Formato: Articulo
Lenguaje:Inglés
Publicado: 2006
Materias:
Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/83049
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-83049
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
spellingShingle Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
Reggiani, Paula Cecilia
Hereñú, Claudia Beatriz
Rimoldi, Omar Jorge
Brown, Oscar Alfredo
Pléau, J. M.
Dardenne, M.
Goya, Rodolfo Gustavo
Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
topic_facet Bioquímica
Thymulin
Fluorescent fusion protein
Thymectomized animals
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.
format Articulo
Articulo
author Reggiani, Paula Cecilia
Hereñú, Claudia Beatriz
Rimoldi, Omar Jorge
Brown, Oscar Alfredo
Pléau, J. M.
Dardenne, M.
Goya, Rodolfo Gustavo
author_facet Reggiani, Paula Cecilia
Hereñú, Claudia Beatriz
Rimoldi, Omar Jorge
Brown, Oscar Alfredo
Pléau, J. M.
Dardenne, M.
Goya, Rodolfo Gustavo
author_sort Reggiani, Paula Cecilia
title Gene therapy for long-term restoration of circulating thymulin in thymectomized mice and rats
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
publishDate 2006
url http://sedici.unlp.edu.ar/handle/10915/83049
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