Fig. 1

Design and expression of TIGIT-based CSRs, TCR F4 and CD155 ligand.a Schematic representation of the different TIGIT chimeras (as indicated). The amino acid numbering (based on the original protein) is indicated below each segment. b Human PBLs were transduced with the retroviral vectors encoding the indicated constructs. 72 h after transduction, the expression of the transgenes was measured by flow cytometry using antibodies specific for TIGIT (upper panels) and F4-TCR (Vβ12 – lower panels). The dotted line represents the basal endogenous expression in the control population. The percentage of positive cells and the MFI (in brackets) are shown. These results are representative of ten independent experiments with at least eight different donors and the difference between the population transduced and the non-transduced population was found statistically significant (p < 0.05; calculated using a Student’s paired t-test). c CD155 expression by melanoma lines (as indicated on the right side) was assessed by flow cytometry. The CD155 expression levels by native cell lines (left column – “WT”) and by CD155-transduced cell lines (right column – “CD155 tr.”) are shown. These results are representative of four independent experiments and the difference between the CD155-stained population and the control population (isotype-stained – dotted line) was found statistically significant (p < 0.05; calculated using a Student’s paired t-test). d-f Following transduction with TIGIT-28 or a control gene (tr.CD34), we measured the distribution of CD4+/CD8+ cells after a 10-day culture (d). No statistically significant difference was observed between the TIGIT-28 and control populations. These cells were stained also for CD45RO and CCR7 expression to determine the memory phenotype of these different populations (e). EM - Effector memory (CD45RO⁺/CCR7⁻), CM - central memory (CD45RO⁺/CCR7⁺), EMRA - terminally differentiated effector memory cells re-expressing CD45RA (CD45RO⁻/CCR7⁻) or naïve cell population (CD45RO⁺/CCR7⁺) are presented. No significant differences were observed in the distribution of these populations between the different treatments (i.e., TIGIT-28 or controls). These results are representative of three independent experiments with three different donors. f Cell count of these cells following transduction with TIGIT-28 + TCR F4, TCR F4 only or mock transduced was determined at different time points as indicated. No significant differences were observed and these results are representative of three independent experiments with three different donors