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Generation of multi-functional antigen-specific human T-cells by lentiviral TCR gene transfer

Gene Therapy volume 17, pages 721–732 (2010)Cite this article

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Abstract

T-cell receptor (TCR) gene transfer is an attractive strategy to generate antigen-specific T-cells for adoptive immunotherapy of cancer and chronic viral infection. However, current TCR gene transfer protocols trigger T-cell differentiation into terminally differentiated effector cells, which likely have reduced ability to mediate disease protection in vivo. We have developed a lentiviral gene transfer strategy to generate TCR-transduced human T-cells without promoting T-cell differentiation. We found that a combination of interleukin-15 (IL15) and IL21 facilitated lentiviral TCR gene transfer into non-proliferating T-cells. The transduced T-cells showed redirection of antigen specificity and produced IL2, IFNγ and TNFα in a peptide-dependent manner. A significantly higher proportion of the IL15/IL21-stimulated T-cells were multi-functional and able to simultaneously produce all three cytokines (P<0.01), compared with TCR-transduced T-cells generated by conventional anti-CD3 plus IL2 stimulation, which primarily secreted only one cytokine. Similarly, IL15/IL21 maintained high levels of CD62L and CD28 expression in transduced T-cells, whereas anti-CD3 plus IL2 accelerated the loss of CD62L/CD28 expression. The data demonstrate that the combination of lentiviral TCR gene transfer together with IL15/IL21 stimulation can efficiently redirect the antigen specificity of resting primary human T-cells and generate multi-functional T-cells.

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Acknowledgements

ECM, MP and HJS designed the research, analysed the data and wrote the paper. MP, JT, SX, LG and DE performed the research. MCG, JT and CP contributed new reagents. MC and DH analysed data. Grant Support: Leukaemia Research, Medical Research Council.

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Authors and Affiliations

  1. Department of Immunology, Division of Infection and Immunity, University College London, Royal Free Hospital, London, UK

    M Perro, J Tsang, S-A Xue, M Cesco-Gaspere, C Pospori, L Gao, D Hart, H Stauss & E C Morris

  2. Division of Infection and Immunity, UCL/MRC Centre for Medical Molecular Virology, University College London, London, UK

    J Tsang, D Escors & M Collins

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Correspondence to E C Morris.

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Perro, M., Tsang, J., Xue, SA. et al. Generation of multi-functional antigen-specific human T-cells by lentiviral TCR gene transfer. Gene Ther 17, 721–732 (2010). https://doi.org/10.1038/gt.2010.4

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