Fig. 2

Akt inhibition generates CD8 + CD19CAR T cell populations with memory-like characteristics. CD8 + CD19CAR T cells that had been cultured in the presence and absence of 1 μM Akti were stained with biotinylated Erbitux (cetuximab), followed by streptavidin-PE for CAR detection and antibodies against CD8, CD62L, CD28, KLRG1, CD57, Tim3, PD1, and LAG3. a Percentages of CD62L+ and CAR + CD62L+ double positive cells are depicted on the basis of the gating of isotype-stained cells. b Percentages of CD62L+ and CAR + CD62L+ double positive cells from four different donors are presented. c Expression and percentages of CD62L and CD28 after gating on CAR+ cells are presented. d Percentages of CD62L + CD28 + CAR+ cells after gating on CAR+ cells. Data from four different donors are presented. e Mean percentages of immunoreactive cells + SEM from 4 different donors are presented. f In vitro expanded CD8 + CD19CAR+ T cells were collected. Relative telomere length of T cells was determined with Telomere PNA detection kit and normalized against 1301 cell line as control. Means + SEMs of duplicates from three different donors are depicted. RTL = (mean FL1 sample cells with probe-mean FL1 sample cells without probe) × DNA index of control cells ×100/(mean FL1 control cells with probe-mean FL1 control cells without probe) × DNA index of sample cells