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CD34-based enrichment of genetically engineered human T cells for clinical use results in dramatically enhanced tumor targeting

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Abstract

Objective clinical responses can be achieved in melanoma patients by infusion of T cell receptor (TCR) gene transduced T cells. Although promising, the therapy is still largely ineffective, as most patients did not benefit from treatment. That only a minority of the infused T cells were genetically modified and that these were extensively expanded ex vivo may have prevented their efficacy. We developed novel and generally applicable retroviral vectors that allow rapid and efficient selection of T cells transduced with human TCRs. These vectors encode two TCR chains and a truncated CD34 molecule (CD34t) in a single mRNA transcript. Transduced T cells were characterized and the effects of CD34-based enrichment of redirected T cells were evaluated. Both CD8+ and CD4+ T cells could be transduced and efficiently co-expressed all introduced transgenes on their surface. Importantly, more than fivefold enrichment of both the frequency of transduced cells and the specific anti-tumor reactivity of the effector population could be achieved by magnetic beads-based enrichment procedures readily available for clinical grade hematopoietic stem cell isolation. This CD34-based enrichment technology will improve the feasibility of adoptive transfer of clinically relevant effectors. In addition to their enhanced tumor recognition, the enriched redirected T cells may also show superior reactivity and persistence in vivo due to the high purity of transduced cells and the shortened ex vivo culture.

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Abbreviations

CTL:

Cytolytic T lymphocytes

PBMC:

Peripheral blood mononuclear cells

TCR:

T cell receptor

IFN:

Interferon

FACS:

Fluorescence-activated cell sorting

ELISA:

Enzyme-linked immunosorbent assay

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Acknowledgments

The work was supported by Abney Foundation postdoctoral scholarship (HN), NIH Grant R01CA104947 (MIN), laboratory start-up funds and American Cancer Society Institutional Research Grant 97-219-08 (YZ) and RBC Research Scholars Program (AT). The authors are also grateful to the MUSC Center for Cellular Therapy for their invaluable support. The Center for Cellular Therapy is supported in part by the Clinical and Translational Science Award support grant (UL 1 RR029882) and the Hollings Cancer Center. We thank Georgiana Onicescu for providing biostatistics expertise and Dr. Toshio Kitamura for providing PlatA cells. The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Division of General Surgery, Department of Surgery, Medical University of South Carolina, Charleston, SC, USA

    Håkan Norell, Telma Martins da Palma, Aaron Lesher, Anquanette Temple & Michael I. Nishimura

  2. Division of Transplantation, Department of Surgery, Medical University of South Carolina, Charleston, SC, USA

    Yi Zhang & Yueying Liu

  3. Section of General Surgery, Department of Surgery, University of Chicago, Chicago, IL, USA

    James McCracken, Jeffrey J. Roszkowski, Glenda G. Callender & José Guevara-Patiño

  4. Department of Surgery, Duke University Medical Center, Durham, NC, USA

    Timothy Clay

  5. Lentigen Corporation, Gaithersburg, MD, USA

    Rimas Orentas

  6. Department of Surgery, Hollings Cancer Center, Medical University of South Carolina, HO503, 86 Jonathan Lucas Street, PO Box 250613, Charleston, SC, 29425, USA

    Håkan Norell

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  1. Håkan Norell
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  2. Yi Zhang
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Correspondence to Håkan Norell.

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Norell, H., Zhang, Y., McCracken, J. et al. CD34-based enrichment of genetically engineered human T cells for clinical use results in dramatically enhanced tumor targeting. Cancer Immunol Immunother 59, 851–862 (2010). https://doi.org/10.1007/s00262-009-0810-8

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  • DOI: https://doi.org/10.1007/s00262-009-0810-8

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