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Gene Ther. 2014 May;21(5):533-8. doi: 10.1038/gt.2014.25. Epub 2014 Apr 3.

T-cell receptor transfer into human T cells with ecotropic retroviral vectors.

Author information

1
III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany.
2
TRON (Translational Oncology at the University Medical Center), Johannes Gutenberg University Mainz, Mainz, Germany.
3
1] III Medical Clinic at the University Medical Center of the Johannes Gutenberg University Mainz, Mainz, Germany [2] TRON (Translational Oncology at the University Medical Center), Johannes Gutenberg University Mainz, Mainz, Germany.

Abstract

Adoptive T-cell transfer for cancer immunotherapy requires genetic modification of T cells with recombinant T-cell receptors (TCRs). Amphotropic retroviral vectors (RVs) used for TCR transduction for this purpose are considered safe in principle. Despite this, TCR-coding and packaging vectors could theoretically recombine to produce replication competent vectors (RCVs), and transduced T-cell preparations must be proven free of RCV. To eliminate the need for RCV testing, we transduced human T cells with ecotropic RVs so potential RCV would be non-infectious for human cells. We show that transfection of synthetic messenger RNA encoding murine cationic amino-acid transporter 1 (mCAT-1), the receptor for murine retroviruses, enables efficient transient ecotropic transduction of human T cells. mCAT-1-dependent transduction was more efficient than amphotropic transduction performed in parallel, and preferentially targeted naive T cells. Moreover, we demonstrate that ecotropic TCR transduction results in antigen-specific restimulation of primary human T cells. Thus, ecotropic RVs represent a versatile, safe and potent tool to prepare T cells for the adoptive transfer.

PMID:
24694535
DOI:
10.1038/gt.2014.25
[Indexed for MEDLINE]

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