Journal of Molecular Medicine

, Volume 86, Issue 5, pp 573–583

Enhanced functionality of T cell receptor-redirected T cells is defined by the transgene cassette

  • Matthias Leisegang
  • Boris Engels
  • Peter Meyerhuber
  • Elisa Kieback
  • Daniel Sommermeyer
  • Shao-An Xue
  • Simone Reuß
  • Hans Stauss
  • Wolfgang Uckert
Original Article

DOI: 10.1007/s00109-008-0317-3

Cite this article as:
Leisegang, M., Engels, B., Meyerhuber, P. et al. J Mol Med (2008) 86: 573. doi:10.1007/s00109-008-0317-3

Abstract

The transfer of T cell receptor (TCR) genes allows to endow T cells with a new antigen specificity. For clinical applications of TCR-redirected T cells, efficient functional expression of the transgenic TCR is a key prerequisite. Here, we compared the influence of the transgene cassette on the expression and function of the murine TCR P14 (recognizing a LCMV gp33 epitope) and the human TCR WT-1 (recognizing an epitope of the tumor-associated antigen WT-1). We constructed different vectors, in which TCRα- and β-chain genes were either (a) linked by an internal ribosomal entry site (IRES), (b) combined by a 2A peptide, or (c) introduced into two individual retroviral constructs. While in a TCR-deficient T cell line TCR P14 was expressed equally well by all constructs, we found that IRES- but not 2A-employing TCR expression is hampered in a TCR-bearing cell line and in primary murine T cells where the transgenic TCR has to compete with endogenous TCR chains. Similarly, 2A-linked TCR WT-1 genes yielded highest expression and function as measured by tetramer binding and peptide-specific IFN-γ secretion. Differences in expression were independent of copy number integration as shown by real-time PCR. Thus, linking TCRα- and β-chain genes by a 2A peptide is superior to an IRES for TCR expression and T cell function.

Keywords

T cell avidity T cell receptor Retroviral vector IRES 2A peptide 

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Matthias Leisegang
    • 1
  • Boris Engels
    • 2
  • Peter Meyerhuber
    • 3
  • Elisa Kieback
    • 1
  • Daniel Sommermeyer
    • 1
  • Shao-An Xue
    • 4
  • Simone Reuß
    • 1
  • Hans Stauss
    • 4
  • Wolfgang Uckert
    • 1
    • 3
  1. 1.Max-Delbrück-Center for Molecular MedicineBerlinGermany
  2. 2.Department of Pathology and Committee on ImmunologyUniversity of ChicagoChicagoUSA
  3. 3.Institute of BiologyHumboldt UniversityBerlinGermany
  4. 4.Department of Immunology and Molecular Pathology, Royal Free HospitalUniversity College LondonLondonUK

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