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Functional impairment of human T-lymphocytes following PHA-induced expansion and retroviral transduction: implications for gene therapy

Gene Therapy volume 9, pages 1359–1368 (2002)Cite this article

Abstract

The immune function of retrovirus-mediated gene modified (GM) T cells is critical for a beneficial effect to follow their adoptive transfer into patients. Recent clinical data show that GM T cells expanded with PHA have reduced function in vivo. However, little functional analysis of PHA stimulation is available. Our results show that expansion of T cells with PHA impairs their ability to respond (proliferation, cytotoxicity and IFNγ and perforin expression) to allogeneic stimulation or viral antigens in vitro. Conversely, CD3/CD28-based protocols can preserve this immune function. Retroviral transduction did not alter the functional profile induced by polyclonal stimulation. We investigated the mechanisms leading to this functional effect, and identified differential effects of PHA and CD3/CD28 on the distribution of CCR7/CD45RA T cell functional subsets, which may explain the functional differences observed. While CD3/CD28 stimulation parallels the lineage differentiation pattern induced by antigens in physiological conditions, PHA induces a skewed distribution of the CCR7/CD45RA functional T cell subsets, with near disappearance of the subpopulations that display the effector phenotype. Overall, this study demonstrates a functional disadvantage for transduction protocols based on PHA, uncovers mechanisms that may explain this functional effect, and provides us with information to design and select transduction protocols with an improved functional outcome.

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Acknowledgements

We thank Professor Y-J Bignon (Centre Jean Perrin, France) and Dr G Dranoff (Dana-Farber Cancer Institute, Boston, MA, USA) for generously sharing reagents, and Dr David A Frank (Dana-Farber Cancer Institute, Boston, MA, USA) and Dr SGE Marsh (The Anthony Nolan Research Institute) for critical reading and stimulating discussion on the manuscript.

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Authors and Affiliations

  1. The Anthony Nolan Research Institute, London, UK

    R F Duarte, F E Chen & J A Madrigal

  2. Department of Haematology, Royal Free and UCL School of Medicine, London, UK

    R F Duarte, F E Chen, M W Lowdell, M N Potter, H G Prentice & J A Madrigal

  3. Ciemat, Madrid, Spain

    M L Lamana

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Duarte, R., Chen, F., Lowdell, M. et al. Functional impairment of human T-lymphocytes following PHA-induced expansion and retroviral transduction: implications for gene therapy. Gene Ther 9, 1359–1368 (2002). https://doi.org/10.1038/sj.gt.3301807

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