Figure 7. p53 inhibitor treated TCR transduced T cells exhibit increased effector function and persistence.

Human peripheral blood T cells from normal healthy individuals were retro-virally transduced with melanoma epitope tyrosinase reactive TIL1383I TCR and were either left untreated or were pretreated for an hour with p53 inhibitors Pifithrin-α (30μM) or Pifithrin-μ (10μM) before TCR restimulation with tyrosinase peptide pulsed T2-A2 cells for 4-6 hrs to analyze: A) Glucose uptake using 2NBDG assay, B) Cytokine secretion by intracellular IFN-γ staining, and C) Susceptibility to AICD by Annexin V staining (*p<0.01). Bar diagram on right of each overlay represent cumulative data from different experiments. D) Ten million human T cells engineered with tyrsoinase reactive TIL1383I TCR were either untreated or pretreated with a combination of p53 inhibitors Pifithrin-α and Pifithrin-μ and adoptively transferred to NSG-A2 mice. Peripheral blood and spleens of recipient mice were stained for human Vβ12, human CD8 and CD4 for tracking the persistence of the transferred cells. Data was acquired using FACS. Numerical value is the average from three mice in similar groups.