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Clinical-scale selection and viral transduction of human naïve and central memory CD8+ T cells for adoptive cell therapy of cancer patients

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Abstract

The adoptive transfer of lymphocytes genetically engineered to express tumor-specific antigen receptors is a potent strategy to treat cancer patients. T lymphocyte subsets, such as naïve or central memory T cells, selected in vitro prior to genetic engineering have been extensively investigated in preclinical mouse models, where they demonstrated improved therapeutic efficacy. However, so far, this is challenging to realize in the clinical setting, since good manufacturing practices (GMP) procedures for complex cell sorting and genetic manipulation are limited. To be able to directly compare the immunological attributes and therapeutic efficacy of naïve (TN) and central memory (TCM) CD8+ T cells, we investigated clinical-scale procedures for their parallel selection and in vitro manipulation. We also evaluated currently available GMP-grade reagents for stimulation of T cell subsets, including a new type of anti-CD3/anti-CD28 nanomatrix. An optimized protocol was established for the isolation of both CD8+ TN cells (CD4−CD62L+CD45RA+) and CD8+ TCM (CD4−CD62L+CD45RA−) from a single patient. The highly enriched T cell subsets can be efficiently transduced and expanded to large cell numbers, sufficient for clinical applications and equivalent to or better than current cell and gene therapy approaches with unselected lymphocyte populations. The GMP protocols for selection of TN and TCM we reported here will be the basis for clinical trials analyzing safety, in vivo persistence and clinical efficacy in cancer patients and will help to generate a more reliable and efficacious cellular product.

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Acknowledgments

This work was supported by grants from ATTRACT European Consortium (FP7-PEOPLE-ITN-2008, Grant Agreement Number 238778); Anna Casati was recipient of a Marie Curie fellowship.

Conflict of interest

Mario Assenmacher is employee of Miltenyi Biotec GmbH and Alexander Scheffold works as a consultant for Miltenyi Biotec GmbH. All other authors do not have any conflicts to disclose.

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Authors and Affiliations

  1. Research and Development, Miltenyi Biotec GmbH, Bergisch Gladbach, Germany

    Anna Casati & Mario Assenmacher

  2. Surgery Branch, National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA

    Azam Varghaei-Nahvi, Steven Alexander Feldman, Steven Aaron Rosenberg & Mark Edward Dudley

  3. Department of Rheumatology and Clinical Immunology, Charité, Universitätsmedizin Berlin, Charitéplatz 1, 10117, Berlin, Germany

    Alexander Scheffold

  4. German Rheumatism Research Centre (DRFZ) Berlin, Leibniz Association, Berlin, Germany

    Alexander Scheffold

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  1. Anna Casati
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  2. Azam Varghaei-Nahvi
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  3. Steven Alexander Feldman
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  7. Alexander Scheffold
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Correspondence to Alexander Scheffold.

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Casati, A., Varghaei-Nahvi, A., Feldman, S.A. et al. Clinical-scale selection and viral transduction of human naïve and central memory CD8+ T cells for adoptive cell therapy of cancer patients. Cancer Immunol Immunother 62, 1563–1573 (2013). https://doi.org/10.1007/s00262-013-1459-x

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  • DOI: https://doi.org/10.1007/s00262-013-1459-x

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