Figure 3.

Anti-tumoral Activity of In Vivo-Generated CAR T Cells

(A) Experimental layout. 1 × 10⁵ Nalm6-Luc tumor cells were i.v. injected into NSG mice. Five days later, 5 × 10⁶ activated human PBMCs were i.v. administered, and finally a day later the mice were i.v. injected with either PBS (control), or 1 × 10¹¹ particles of CD4-LV, or 2.5 × 10¹¹ particles of CD8-LV, or 5 × 10¹⁰ particles of CD4-LV and 1.25 × 10¹¹ particles of CD8-LV (MIX) encoding for CD19-CAR. 14 and 21 days later mice were sacrificed and organs were harvested. (B) Ventral view of all mice included in the experiment recorded at the indicated time points after PBS (control) or vector (CD4-LV, CD8-LV, MIX) administration. Each column represents the bioluminescence imaging (BLI) development of one individual mouse over time. Mice sacrificed at day 14 are indicated by “X” in the day 20 panel. (C) Kinetics of quantified luciferase signals over time is shown as logarithm of the total flux (photons [p]/s). The dotted line indicates the time point of vector administration. Data represent mean ± SEM for all groups (control, n = 6; CD4-LV, n = 7; CD8-LV, n = 6; MIX, n = 7). Statistical significance was determined using a two-way ANOVA with Turkey’s multiple comparisons test. ∗∗∗∗p < 0.0001, comparing different groups to control; ⁺⁺⁺⁺p < 0.0001, comparing CD8-LV to CD4-LV; ^#p < 0.05, comparing MIX to CD4-LV.