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

Eur J Immunol. 2011 May;41(5):1288-97. doi: 10.1002/eji.200940233. Epub 2011 Apr 13.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adoptive Transfer
  • CD28 Antigens / immunology
  • CD28 Antigens / metabolism
  • CD3 Complex / genetics
  • CD3 Complex / immunology
  • CD8 Antigens / immunology
  • CD8 Antigens / metabolism
  • Flow Cytometry
  • Genetic Therapy
  • Histocompatibility Antigens Class I
  • Humans
  • Immunity, Cellular
  • Immunological Synapses / physiology*
  • Jurkat Cells
  • Leukocyte Common Antigens / immunology
  • Leukocyte Common Antigens / metabolism
  • Membrane Microdomains / metabolism
  • Receptor-CD3 Complex, Antigen, T-Cell / genetics
  • Receptor-CD3 Complex, Antigen, T-Cell / immunology*
  • Receptors, Antigen, T-Cell, alpha-beta / genetics
  • Receptors, Antigen, T-Cell, alpha-beta / immunology*
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism
  • Transgenes


  • CD28 Antigens
  • CD3 Complex
  • CD3 antigen, zeta chain
  • CD8 Antigens
  • Histocompatibility Antigens Class I
  • Receptor-CD3 Complex, Antigen, T-Cell
  • Receptors, Antigen, T-Cell, alpha-beta
  • Leukocyte Common Antigens