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Pharmacological control of antigen responsiveness in genetically modified T lymphocytes.
Alvarez-Vallina, L; Agha-Mohammadi, S; Hawkins, R E; Russell, S J.
Afiliación
  • Alvarez-Vallina L; Center for Protein Engineering, Medical Research Council Center, Cambridge, United Kingdom. lav@mrc-lmb.cam.ac.uk
J Immunol ; 159(12): 5889-95, 1997 Dec 15.
Article en En | MEDLINE | ID: mdl-9550385
A chimeric TCR gene, comprising an anti-hapten single-chain Ab variable fragment fused to the transmembrane and cytoplasmic regions of the human TCR zeta-chain, was used to determine whether the tetracycline-regulatable system could be used to regulate gene expression in T cells. Jurkat T cells were stably transfected with a single vector encoding the tetracycline trans-activator protein, controlled by a constitutive promoter, and the chimeric TCR, under the control of a trans-activator protein-responsive promoter. In the absence of tetracyclines, the transfected T cells were shown to express the chimeric receptor on the cell surface and could be activated by its cognate Ag, leading to the secretion of IL-2. When the cells were exposed to increasing concentrations of tetracyclines, surface expression of the chimeric receptor was suppressed in a dose-dependent manner, and this suppression was sufficient to result in complete loss of responsiveness to the targeted Ag. Prolonged suppression of trans-gene expression for up to 7 days was observed after doxycycline was removed from the cultures, but eventual recovery of surface expression was complete, and the absolute time to recovery was directly proportional to the initial concentration of the drug. Pharmacologic control of trans-gene expression in gene-modified T cells will not only facilitate new approaches to the study of different aspects of T cell biology, but will also provide the basis for new gene therapy strategies.
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Tetraciclina / Activación de Linfocitos / Linfocitos T / Vectores Genéticos / Antígenos Límite: Humans Idioma: En Revista: J Immunol Año: 1997 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos
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Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Tetraciclina / Activación de Linfocitos / Linfocitos T / Vectores Genéticos / Antígenos Límite: Humans Idioma: En Revista: J Immunol Año: 1997 Tipo del documento: Article País de afiliación: Reino Unido Pais de publicación: Estados Unidos