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Modulating the differentiation status of ex vivo-cultured anti-tumor T cells using cytokine cocktails

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Abstract

The genetic modification of CD8+ T cells using anti-tumor T-cell receptors (TCR) or chimeric antigen receptors is a promising approach for the adoptive cell therapy of patients with cancer. We previously developed a simplified method for the clinical-scale generation of central memory-like (Tcm) CD8+ T cells following transduction with lentivirus encoding anti-tumor TCR and culture in the presence of IL-2. In this study, we compared different cytokines or combinations of IL-2, IL-7, IL-12, IL-15, and IL-21 to expand genetically engineered CD8+ T cells. We demonstrated that specific cytokine combinations IL-12 plus IL-7 or IL-21 for 3 days followed by withdrawal of IL-12 yielded the phenotype of CD62LhighCD28high CD127highCD27highCCR7high, which is associated with less-differentiated T cells. Genes associated with stem cells (SOX2, NANOG, OCT4, and LIN28A), were also up-regulated by this cytokine cocktail. Moreover, the use of IL-12 plus IL-7 or IL-21 yielded CD8 T cells showing enhanced persistence in the NOD/SCID/γc−/− mouse model. This defined cytokine combination could also alter highly differentiated TIL from melanoma patients into cells with a less-differentiated phenotype. The methodology that we developed for generating a less-differentiated anti-tumor CD8+ T cells ex vivo may be ideal for the adoptive immunotherapy of cancer.

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Acknowledgments

We thank Arnold Mixon and Shawn Farid in the FACS laboratory and all members in the TIL laboratory at the Surgery Branch for providing technical support and maintenance of PBL and tumor cells from patients. This work is supported by the Intramural Research Program of the National Institute of Health, National Cancer Institute, Center for Cancer Research.

Conflict of interest

The authors declare that they have no competing financial interests.

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Author notes

  1. Shicheng Yang

    Present address: Department of Surgery, Duke University Medical Center, Durham, NC, 27710, USA

Authors and Affiliations

  1. Surgery Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, 10 Center Drive, Building 10, CRC 3 W-3864, Bethesda, MD, 20892, USA

    Shicheng Yang, Yun Ji, Luca Gattinoni, Ling Zhang, Zhiya Yu, Nicholas P. Restifo, Steven A. Rosenberg & Richard A. Morgan

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  1. Shicheng Yang
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Correspondence to Richard A. Morgan.

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Yang, S., Ji, Y., Gattinoni, L. et al. Modulating the differentiation status of ex vivo-cultured anti-tumor T cells using cytokine cocktails. Cancer Immunol Immunother 62, 727–736 (2013). https://doi.org/10.1007/s00262-012-1378-2

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  • DOI: https://doi.org/10.1007/s00262-012-1378-2

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