A Self-Inactivating Retrovector Incorporating the IL-2 Promoter for Activation- Induced Transgene Expression in Genetically Engineered T-Cells - Genetic Engineering: Recent Developments in Applications - page 232

Figure 4. Activation-induced EGFP expression in Jurkat P116 cells transduced with SINIL-2pr retroparticles.
A-D. Flow cytometry analysis of mean EGFP expression in Jurkat cells deficient for ZAP-70 (Jurkat P116)
transduced with SINIL-2pr retroparticles relative to null cells. Expression was measured at ~48 hr post-activation
with 1 µ M ionomycin and 10 ng/ml PMA relative to untreated counterparts. E. Cyclosporin A-sensitive
induction of EGFP expression in Jurkat P116 cells transduced with SINIL-2pr. Co-stimulation of the SINIL-
2pr- gene modified Jurkat P116 T-cells with 1 µ M ionomycin and 10 ng/ml PMA resulted in a 3.4 ± 0.4 - fold
increase in relative mean EGFP expression measured at ~48 hrs (P = 0.029). This activation-induced EGFP
expression was abrogated when the cells were pretreated for ~30 minutes with 300 nM CsA.
discussion
In this study, we first compared the transcriptional potency of two T-cell activa-
tion-dependent promoters, the IL-2 promoter and a synthetic promoter com-
posed of 3 repeats of the NFAT element following pharmacological stimulation
of transiently transfected T-cells. Plasmid vectors incorporating one of the two
promoters were used to transfer luciferase reporter expression into Jurkat T-cells