In vitro functionality of NFAT-inducible lentiviral vector. (A) Sorted extGLuc⁺ and extGLuc⁻ primary T cells from chimeric B6 mice were stimulated with PMA/Ionomycin or αCD3/CD28 in the presence or absence of Cyclosporine A (CsA). ExtGLuc and NFAT-induced CBRLuc activity were assessed using the IVIS bioluminescence imaging system. A representative image from 1 of 2 identical experiments is shown. (B) Activation marker (CD25, CD44, and CD62L) expression were determined for primary transgenic T cells after stimulation with PMA (100 ng/mL)/ionomycin (1500 ng/mL) and αCD3/CD28 for 16 hours in the presence or absence of CsA (300 ng/mL) and analyzed by flow cytometry. (C) ExtGLuc⁺ T cells from chimeric B6 were challenged with wild-type BALB/c (allogeneic) or B6 (syngeneic) antigen presenting cells plus αCD28 antibody (c = 2 μg/mL) for 72 hours. The extGluc and CBRLuc signal activity (photons/s) were quantified. Data shown represent average of 2 identical experiments.