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

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.

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Acknowledgements

We thank Ines Eiser, Aileen Laubenheimer, Tina Hempel and Kathleen Hobohm for excellent technical assistance, Sebastian Kreiter for proofreading the manuscript and Stephen Reece for native-speaker review.

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Correspondence to U Sahin.

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ÖT and US are inventors on patent applications, which cover optimization of RNA-based therapeutics. The remaining authors declare no conflict of interest.

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Supplementary Information accompanies this paper on Gene Therapy website

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Koste, L., Beissert, T., Hoff, H. et al. T-cell receptor transfer into human T cells with ecotropic retroviral vectors. Gene Ther 21, 533–538 (2014). https://doi.org/10.1038/gt.2014.25

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