Figure 2.

LunX Antigen Induces Expression of the Immune-Function Molecules of CAR^LunX T Cells

(A) Microscopic appearance of enhanced GFP (EGFP) expressed by lentivirus-infected human primary T cells. Scale bar, 50 μm. (B) Expression of chimeric s-35-8 scFv on the surface of human primary T cells transduced with the LunX-CAR construct was measured by flow cytometry after cells had been stained with an anti-myc antibody or IgG1 isotype control. Data are representative of three experiments with similar results. (C) Immunofluorescence staining for c-myc tag and EGFP of human primary T cells transduced with the LunX-CAR construct. Scale bar, 5 μm. (D)The graphical representation of experimental protocol in (E). (E) Indirect ELISAs quantifying production of the cytokines IFN-γ, IL-2, or TNF-α in supernatants from LunX CAR T cells and CD19 CAR T cells cultured on peptide-bound plates for 24 h. LunX-antigen peptides were plated at 1,000 ng/mL. Antibody against OKT3 (10 μg/mL) but not transfected by lentivirus was used as a control stimulant of T cells. n = 3, and results are representative of three independent experiments. Data in (E) are the mean ± SEM. Unpaired t test, ∗∗∗∗p < 0.0001.