Pharmacological Control of Antigen Responsiveness in Genetically Modified T Lymphocytes

Luis Alvarez-Vallina, Siamak Agha-Mohammadi, Robert E. Hawkins, Stephen J. Russell

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

A chimeric TCR gene, comprising an anti-hapten single-chain Ab variable fragment fused to the transmembrane and cytoplasmic regions of the human TCR ζ-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.

Original languageEnglish (US)
Pages (from-to)5889-5895
Number of pages7
JournalJournal of Immunology
Volume159
Issue number12
StatePublished - Dec 15 1997

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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