Efficient transfer of HTLV-1 tax gene in various primary and immortalized cells using a flap lentiviral vector.
Human T cell leukemia virus type 1 (HTLV-1) causes two major diseases: adult T-cell leukemia-lymphoma and tropical spastic paraparesis/HTLV-1 associated myelopathy (TSP/HAM). In order to understand the involvement of Tax protein in HTLV-1 pathogenesis, we constructed a HIV-1 based lentiviral vector containing the central DNA flap sequence and either the green fluorescent protein (GFP) or the HTLV-1 tax genes. Using these vectors, GFP and tax genes were introduced in several primary and immortalized cells of endothelial, lymphoid, astrocytic or macrophagic origin. As assessed by GFP expression, up to 100% efficiency of transduction was obtained for all cell types tested. Tax expression was detected by Western blot and immuno-fluorescence in the transduced cells. After transduction, the Tax transcriptional activity was confirmed by the transactivation of HTLV-1 LTR-lacZ or HTLV-1 LTR-GFP reporter genes. Increased CD25 and HLA DR expression was observed in human peripheral blood lymphocytes transduced with the Tax vector. These results indicate that both pathways of Tax transactivation, CREB (viral LTR) and NF-kappa B (CD25 and HLA DR), are functional after transduction by TRIP Tax vector. Therefore, this vector provides a useful tool for investigating the role of the Tax viral protein in the pathogenesis of diseases linked to HTLV-1 infection.
Royer-Leveau
Christelle
C
Unité d'Epidémiologie et Physiopathologie des Virus Oncogènes, Institut Pasteur, 25-28 rue du Dr Roux, 75724 Paris,Cedex 15 France.
Mordelet
Elodie
E
Delebecque
Frédéric
F
Gessain
Antoine
A
Charneau
Pierre
P
Ozden
Simona
S
D002461:Cell Line, Transformed; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics; D016356:Gene Products, tax / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D015497:HIV-1 / Q000235:genetics; D015490:HTLV-I Infections / Q000503:physiopathology / Q000821:virology; D015368:Human T-lymphotropic virus 1 / Q000378:metabolism; D006801:Humans; D007763:Lac Operon / Q000235:genetics / Q000502:physiology; D016086:Lentivirus / Q000235:genetics*; D007963:Leukocytes, Mononuclear / Q000821:virology; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D014161:Transduction, Genetic
D004279:DNA, Viral; D016356:Gene Products, tax; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins