T-cell synapse formation depends on antigen recognition but not CD3 interaction: Studies with TCR:ζ, a candidate transgene for TCR gene therapy

Authors

  • János Roszik,

    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Zsolt Sebestyén,

    1. Laboratory of Experimental Tumor Immunology, Department of Medical Oncology, Erasmus Medical Center – Daniel den Hoed Cancer Center, Rotterdam, The Netherlands
    Search for more papers by this author
    • These authors contributed equally to this work.

  • Coen Govers,

    1. Laboratory of Experimental Tumor Immunology, Department of Medical Oncology, Erasmus Medical Center – Daniel den Hoed Cancer Center, Rotterdam, The Netherlands
    Search for more papers by this author
  • Yakir Guri,

    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    Search for more papers by this author
  • Árpád Szöőr,

    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    Search for more papers by this author
  • Zsuzsanna Pályi-Krekk,

    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    Search for more papers by this author
  • György Vereb,

    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    Search for more papers by this author
  • Peter Nagy,

    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    Search for more papers by this author
  • János Szöllősi,

    Corresponding author
    1. Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    2. Cell Biology and Signaling Research Group of the Hungarian Academy of Sciences, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary
    • Department of Biophysics and Cell Biology, Medical and Health Science Center, University of Debrecen, P. O. Box 39, Nagyerdei krt. 98, H-4012 Debrecen, Hungary Fax: +36-52-532-201
    Search for more papers by this author
  • Reno Debets

    Corresponding author
    1. Laboratory of Experimental Tumor Immunology, Department of Medical Oncology, Erasmus Medical Center – Daniel den Hoed Cancer Center, Rotterdam, The Netherlands
    • Laboratory of Experimental Tumor Immunology, Department of Medical Oncology, Erasmus Medical Center-Daniel den Hoed Cancer Center, Groene Hilledijk 301 3175EA, Rotterdam Fax: +31-107041005
    Search for more papers by this author

Abstract

T-cell receptors (TCRs) can be genetically modified to improve gene-engineered T-cell responses, a strategy considered critical for the success of clinical TCR gene therapy to treat cancers. TCR:ζ, which is a heterodimer of TCRα and β chains each coupled to complete human CD3ζ, overcomes issues of mis-pairing with endogenous TCR chains, shows high surface expression and mediates antigen-specific T-cell functions in vitro. In the current study, we further characterized TCR:ζ in gene-engineered T cells and assessed whether this receptor is able to interact with surface molecules and drive correct synapse formation in Jurkat T cells. The results showed that TCR:ζ mediates the formation of synaptic areas with antigen-positive target cells, interacts closely with CD8α and MHC class I (MHCI), and co-localizes with CD28, CD45 and lipid rafts, similar to WT TCR. TCR:ζ did not closely associate with endogenous CD3ε, despite its co-presence in immune synapses, and TCR:ζ showed enhanced synaptic accumulation in T cells negative for surface-expressed TCR molecules. Notably, synaptic TCR:ζ demonstrated lowered densities when compared with TCR in dual TCR T cells, a phenomenon that was related to both extracellular and intracellular CD3ζ domains present in the TCR:ζ molecule and responsible for enlarged synapse areas.

Ancillary