A retroviral vector that directs simultaneous expression of α and β T cell receptor genes

Robert J. Pogulis, Larry R Pease

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

The transfer of α/β T cell receptor (TCR) genes into T lymphocytes or their precursors could provide a means to increase frequency of tumor- or pathogen-specific cytotoxic T lymphocytes. To begin to address this possibility, we have used class I MHC-restricted α/β TCR cDNAs to develop a retroviral TCR expression vector, α- and β-chain cDNAs were inserted into a derivative of the LN series of retroviral vectors, with the retroviral LTR directing expression of TCR-β and an internal cytomegalovirus promoter/enhancer driving TCR-α expression. The variable region fragments can be replaced using unique restriction sites that have been introduced into the proximal constant regions. We have used this vector system to transfer two different pairs of α/β TCR genes into an α- and β-chain-deficient T cell hybridoma. TCR- hybridoma cells were transduced by coculture with pools of virus-producing cells, and fluorescence-activated cell sorting was used to enrich for cells expressing the transduced TCR. Transduction with either α/β TCR restores stable, long-lived expression of the α/β TCR complex. TCR-mediated signal transduction is also reconstituted, as demonstrated by the ability of transduced cells to secrete IL-2 following stimulation with Vβ-specific antibodies. Our results suggest that α/β T cell receptor gene transfer could provide a basis for new approaches to immunotherapy, and that further studies examining the in vivo fate of transduced TCR are possible.

Original languageEnglish (US)
Pages (from-to)2299-2304
Number of pages6
JournalHuman Gene Therapy
Volume9
Issue number15
StatePublished - Oct 10 1998

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T-Cell Receptor Genes
T-Cell Antigen Receptor
Hybridomas
Complementary DNA
T-Lymphocytes
Cytotoxic T-Lymphocytes
Coculture Techniques
Cytomegalovirus
Immunotherapy
Interleukin-2
Signal Transduction
Flow Cytometry
Viruses

ASJC Scopus subject areas

  • Genetics

Cite this

A retroviral vector that directs simultaneous expression of α and β T cell receptor genes. / Pogulis, Robert J.; Pease, Larry R.

In: Human Gene Therapy, Vol. 9, No. 15, 10.10.1998, p. 2299-2304.

Research output: Contribution to journalArticle

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