title: Peripheral and mesencephalic transfer of a synthetic gene for the thymic peptide thymulin.
abstract: Thymulin is a thymic peptide with antiinflammatory activity in the brain. We constructed a recombinant adenoviral vector, RAd-FTS, expressing a synthetic DNA sequence encoding met-FTS, a biologically active analog of thymulin and used it for peripheral and central gene transfer in rats. Thymulin concentration in serum and brain tissue was determined by bioassay. Reporter gene expression in the substantia nigra (SN) was quantitated by enzymohistochemistry or fluorescence microscopy using an appropriate image analysis software. A single intramuscular injection (10(8) plaque forming units (pfu)/animal) of RAd-FTS in thymectomized rats (nondetectable serum thymulin) induced supraphysiologic serum thymulin levels for at least 110 days (123+/-22 fg/ml versus 598+/-144 fg/ml in intact and vector-injected rats, respectively). Stereotaxic intranigral injection of RAd-FTS induced steady expression levels of met-FTS for at least 90 days, whereas expression of adenovirally transferred reporter genes coding for green fluorescent protein fused to HSV thymidine kinase (GFP-TK)(fus) or E.coli beta-galactosidase (beta-gal), declined drastically within a month (% transgene expression in the SN on post-injection day 30 relative to day 2 was: 18, <1 and 125%, for beta-gal, (GFP-TK)(fus) and met-FTS, respectively). We conclude that RAd-FTS constitutes a suitable biotechnological tool for the assessment of peripheral and central thymulin gene therapy in animal models of nigral dopaminergic neurodegeneration induced by pro-inflammatory agents.
authors.affiliation: Institute for Biochemical Research-Histology B (INIBIOLP), Faculty of Medicine, UNLP, CC 455, 1900 La Plata, Argentina. gustavmorel@Argentina.com
authors.forename: Gustavo R
authors.initials: GR
authors.lastname: Morel
authors.forename: Oscar A
authors.initials: OA
authors.lastname: Brown
authors.forename: Paula C
authors.initials: PC
authors.lastname: Reggiani
authors.forename: Claudia B
authors.initials: CB
authors.lastname: Hereñú
authors.forename: Enrique L
authors.initials: EL
authors.lastname: Portiansky
authors.forename: Gustavo O
authors.initials: GO
authors.lastname: Zuccolilli
authors.forename: Jean M
authors.initials: JM
authors.lastname: Pléau
authors.forename: Mireille
authors.initials: M
authors.lastname: Dardenne
authors.forename: Rodolfo G
authors.initials: RG
authors.lastname: Goya
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D003952:Diagnostic Imaging / Q000379:methods; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology; D018014:Gene Transfer Techniques; D017930:Genes, Reporter / Q000502:physiology; D005813:Genes, Synthetic / Q000502:physiology; D005822:Genetic Vectors / Q000502:physiology; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D013378:Substantia Nigra / Q000378:metabolism*; D013934:Thymectomy / Q000379:methods; D013935:Thymic Factor, Circulating / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors
chemical_list: D013935:Thymic Factor, Circulating