title: Pre-TCRα supports CD3-dependent reactivation and expansion of TCRα-deficient primary human T-cells.
abstract: Chimeric antigen receptor technology offers a highly effective means for increasing the anti-tumor effects of autologous adoptive T-cell immunotherapy, and could be made widely available if adapted to the use of allogeneic T-cells. Although gene-editing technology can be used to remove the alloreactive potential of third party T-cells through destruction of either the α or β T-cell receptor (TCR) subunit genes, this approach results in the associated loss of surface expression of the CD3 complex. This is nonetheless problematic as it results in the lack of an important trophic signal normally mediated by the CD3 complex at the cell surface, potentially compromising T-cell survival in vivo, and eliminating the potential to expand TCR-knockout cells using stimulatory anti-CD3 antibodies. Here, we show that pre-TCRα, a TCRα surrogate that pairs with TCRβ chains to signal proper TCRβ folding during T-cell development, can be expressed in TCRα knockout mature T-cells to support CD3 expression at the cell surface. Cells expressing pre-TCR/CD3 complexes can be activated and expanded using standard CD3/CD28 T-cell activation protocols. Thus, heterologous expression of pre-TCRα represents a promising technology for use in the manufacturing of TCR-deficient T-cells for adoptive immunotherapy applications.
authors.affiliation: Cellectis Therapeutics , Paris, France.
authors.forename: Román
authors.initials: R
authors.lastname: Galetto
authors.affiliation: Cellectis Therapeutics , Paris, France.
authors.forename: Celine
authors.initials: C
authors.lastname: Lebuhotel
authors.affiliation: Cellectis Therapeutics , Paris, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Poirot
authors.affiliation: Cellectis Therapeutics , Paris, France.
authors.forename: Agnès
authors.initials: A
authors.lastname: Gouble
authors.affiliation: Centro de Biologia Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid , Madrid, Spain.
authors.forename: Maria L
authors.initials: ML
authors.lastname: Toribio
authors.affiliation: Cellectis Therapeutics , Paris, France.
authors.forename: Julianne
authors.initials: J
authors.lastname: Smith
authors.affiliation: Cellectis Therapeutics , Paris, France.
authors.forename: Andrew
authors.initials: A
authors.lastname: Scharenberg