Fig. 6
From: Ex vivo Akt inhibition promotes the generation of potent CD19CAR T cells for adoptive immunotherapy
Akti-treated CD4 + CD19CAR+ and CD8 + CD19CAR T cells exhibit increased anti-tumor efficacy and persistence in vivo. PBMC from healthy donors were activated with CD3/CD28 Dynabeads and transduced the next day with CD19R (EQ):CD28:ζ/EGFRt lentivirus at an MOI of 3. The cultures were supplemented with 50 U/mL rhIL-2 and Akt inhibitor every 48 h for 21 days. Transduced CD19CAR T cells without Akt inhibitor treatment were used as control. Non-transduced mock T cells were used as another type of control. a Percentages of CD8 and CAR positive cells in the input population are presented. b Acute lymphoid leukemic 0.5 × 106 SupB15 cells engineered with GFPffluc were inoculated into NSG mice (N = 3 mice per group) intravenously (i.v) on day -5. After confirmation of the tumor engraftment, 1 × 106 expanded CD19CAR+ T cells were adoptively transferred into tumor-bearing mice intravenously. Tumor signals were monitored by Biophotonic tumor imaging and c the bioluminescence signal was measured as total photon flux normalized for exposure time and surface area and expressed in units of photons (p) per second per cm2 per steradian (sr). d Forty days post adoptive transfer, mice were euthanized, and blood were analyzed after staining with antibodies against human CD45, CD8 and cetuximab for CAR detection. Mean percentages of human CD45 positive and GFP (tumor) negative cells from three mice are presented. e CD8+ and CAR T cells gated on CD45 positive human T cells are depicted