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Retroviral transduction of human peripheral blood lymphocytes with bcl-xL promotes in vitro lymphocyte survival in pro-apoptotic conditions

Gene Therapy volume 9, pages 527–535 (2002)Cite this article

Abstract

The prolonged in vivo survival of genetically modified effector cells is crucial to the success of any (gene-modified) adoptive cellular immunotherapy approach. In cancer clinical trials to date, however, the detection of surviving circulating gene-modified T cells has required highly sensitive techniques. In vitro studies of T cell co-stimulation have shown that up-regulation of the anti-apoptosis gene Bcl-XL by ligation of CD28 promotes T cell survival, but not proliferation. Here we have investigated the ability to modulate resistance to apoptosis and improve cell survival by transducing human peripheral blood lymphocytes using a retroviral vector that expresses Bcl-XL. We show that Jurkat cells transduced with Bcl-XL retrovirus were partially resistant to Fas (CD95) antibody-induced apoptosis. Subsequent in vitro assays with transduced primary human lymphocytes demonstrates that over-expression of Bcl-XL promotes the survival of lymphocytes cultured in the absence of interleukin-2. Activation-induced apoptosis with anti-CD3ɛ antibody, OKT3 is also modulated. Furthermore, Bcl-XL over-expression in human lymphocytes delays the onset of apoptosis induced by long-term co-culture with tumour cell lines. Despite this improved in vitro survival, in a preliminary experiment to assess safety, no signs of malignancy or autoimmunity were observed in NOD/SCID mice injected with Bcl-XL transduced lymphocytes. These results indicate that expression of Bcl-XL in lymphocyte therapy either alone or in conjunction with an additional therapeutic gene could enhance persistence of cells in vivo thereby potentially improving the clinical outcome of adoptive cellular therapy.

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Acknowledgements

Cancer Research UK funds DE, DEG and AON. We thank Mike Hughes and Jeff Barry (Paterson Institute for Cancer Research, Manchester, UK) for their assistance with flow cytometric analysis. This work was also supported by EU Framework Programme V-QLK3-1999-0162.

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  1. CRC Department of Medical Oncology, Paterson Institute for Cancer Research, University of Manchester, Manchester, UK

    D Eaton, D E Gilham, A O'Neill & R E Hawkins

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  1. D Eaton
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  2. D E Gilham
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  3. A O'Neill
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  4. R E Hawkins
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Eaton, D., Gilham, D., O'Neill, A. et al. Retroviral transduction of human peripheral blood lymphocytes with bcl-xL promotes in vitro lymphocyte survival in pro-apoptotic conditions. Gene Ther 9, 527–535 (2002). https://doi.org/10.1038/sj.gt.3301685

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