Figure 1.

Design and expression of NCR2-based chimeras. (A) Schematic representation of the different NCR-2-based chimeric receptors. The amino acid numbering (based on the original protein) is indicated below each segment. (B) OKT3-stimulated human primary PBLs were transduced with the different versions of NCR2-based chimeric antigen receptor or with truncated CD34 (control gene) as indicated. Transgene expression was assessed by flow cytometry. The dotted line represents the staining of the mock-transduced control. The percentage of positive cells and the MFI (in brackets) are shown. These results are representative of six independent experiments with at least four different donors and the difference between the population transduced with NCR2-chimeric molecules and the control population was found statistically significant (p < 0.05; calculated using a Student’s paired t-test). (C) These cells (10⁵) were incubated in a 96-well plate in the presence of plate-bound anti-NCR2 (0.2 μg/well) or OKT3 for 16 h. IFNγ secreted in the coculture supernatant was measured by ELISA.