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TCR Gene Therapy for Cancer

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Gene Therapy of Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2521))

Abstract

The protocol describes the procedure of antigen-specific T cell generation and TCR identification for the use in adoptive T cell therapy. We describe two paths of generating antigen-specific T cells, first, T cell stimulation with autologous dendritic cells pulsed with antigen peptide, second, in vivo T cell stimulation with peptide or DNA by gene gun application in a suitable mouse model followed by in vitro enrichment of peptide-reactive T cells. Peptide-stimulated T cells are sorted by fluorescence-activated cell sorting for CD107α or IFNγ expression and subsequently isolated RNA is used in a 5′ rapid amplification of cDNA ends (RACE )-PCR specific for TCR for TCR chain identification. After retroviral cloning, it is re-expressed on human T cells to test its applicability in adoptive T cell therapy.

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Author information

Authors and Affiliations

  1. Max Delbrück Center for Molecular Medicine in the Helmholtz Association, Berlin, Germany

    Simone Rhein

  2. Charité-Universitätsmedizin, corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany

    Neşe Çakmak-Görür

Authors
  1. Simone Rhein
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  2. Neşe Çakmak-Görür
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Corresponding author

Correspondence to Simone Rhein .

Editor information

Editors and Affiliations

  1. Max-Delbrück-Center for Molecular Medicine, Berlin, Berlin, Germany

    Wolfgang Walther

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© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Rhein, S., Çakmak-Görür, N. (2022). TCR Gene Therapy for Cancer. In: Walther, W. (eds) Gene Therapy of Cancer. Methods in Molecular Biology, vol 2521. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2441-8_6

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  • DOI: https://doi.org/10.1007/978-1-0716-2441-8_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2440-1

  • Online ISBN: 978-1-0716-2441-8

  • eBook Packages: Springer Protocols

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