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. 2016 Aug 22;126(9):3479–3494. doi: 10.1172/JCI86437

Figure 4. JQ1 treatment improves the antitumor activity of chimeric antigen receptor–transduced T cells in a mouse leukemia model.

Figure 4

(A) CD3+ T cells were retrovirally transduced with the anti-CD19 chimeric antigen receptor (CAR) consisting of anti-CD19 scFv, CD28, and CD3z, and the cells were cultured with JQ1 or (–)-JQ1. The drugs were removed on day 14, and the functional properties of the CAR T cells were analyzed. (B) CAR T cells generated as shown in A were serially stimulated with K562 transduced with CD19 (K562-CD19), and the fold expansion of the CD8+ CAR T cells was calculated at the indicated time points (n = 5, paired t test). Error bars indicate the SD. (C and D) JQ1-CAR T and (–)-JQ1-CAR T cells were labeled with CFSE and stimulated with K562-CD19. The relative mean fluorescence intensity of CFSE (C) and cell viability (D) were assessed 4 days following stimulation (n = 5, paired t test). (E) Cytokine secretion profiles of JQ1-CAR T and (–)-JQ1-CAR T cells upon stimulation with K562-CD19 (n = 5, paired t test). (F) CAR T cells were cocultured with the CD19-positive leukemia cell line NALM-6, which expresses EGFP-luciferase fusion protein (NALM6-GL), at the indicated ratio, and the residual NALM6-GL was analyzed by flow cytometry (n = 5, unpaired t test). (G) NSG mice were intravenously injected with NALM6-GL, then with CAR-transduced T cells 14 days after tumor injection. (H) Total photon counts measured from the in vivo bioluminescent imaging of luciferase activity at the indicated time points following infusion of T cells. (I) Kaplan–Meier curve for overall survival of the mice treated with CAR-transduced T cells (n = 10, log-rank test). Representative data from 2 experiments are shown. (J) Frequency of human CD45+CD8+ CAR-transduced T cells in the peripheral blood following T cell injection. (K and L) The persistence of CD45RA+/–CD62L+CCR7+ T cells was monitored in the peripheral blood. The frequencies within the CD8+ CAR+ T cell population (K) and overall peripheral blood cells (L) (n = 10, unpaired t test) are shown. *P < 0.05, **P < 0.01. NS, not significant.