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

• Published in: European journal of immunology 2011-05, Vol.41 (5), p.1288-1297
• Main Authors: Roszik, János, Sebestyen, Zsolt, Govers, Coen, Guri, Yakir, Szor, Árpád, Palyi-Krekk, Zsuzsanna, Vereb, György, Nagy, Peter, Szollsi, János, Debets, Reno
• Format: Article
• Language: English
• Subjects: Gene therapy; Lymphocytes; Medical research; T cell receptors
• ISSN: 0014-2980; 1521-4141
• DOI: 10.1002/eji.200940233

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:[zeta], which is a heterodimer of TCR[alpha] and [beta] chains each coupled to complete human CD3[zeta], 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:[zeta] 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:[zeta] mediates the formation of synaptic areas with antigen-positive target cells, interacts closely with CD8[alpha] and MHC class I (MHCI), and co-localizes with CD28, CD45 and lipid rafts, similar to WT TCR. TCR:[zeta] did not closely associate with endogenous CD3[epsi], despite its co-presence in immune synapses, and TCR:[zeta] showed enhanced synaptic accumulation in T cells negative for surface-expressed TCR molecules. Notably, synaptic TCR:[zeta] demonstrated lowered densities when compared with TCR in dual TCR T cells, a phenomenon that was related to both extracellular and intracellular CD3[zeta] domains present in the TCR:[zeta] molecule and responsible for enlarged synapse areas. [PUBLICATION ABSTRACT]