Fig. 4: ITK inhibitor soquelitinib mitigates exhaustion in CD19 CAR-T cells in vitro.

A–D Summary data showing the percentages of PD-1⁺, TIM3⁺, TIGIT⁺, and LAG3⁺ cells within CD4⁺ and CD8⁺ CD19 CAR-T cell populations after 7-day exposure to soquelitinib (0-40 nM) with anti-CD3/CD28 bead activation and IL-2 (500 IU/mL); n = 4–6. E–H Summary data showing the percentages of LAG3⁺, TIGIT⁺, TIM3⁺, and CD39⁺ cells within CD4⁺ and CD8⁺ CD19 CAR-T cell populations after 7-day exposure to soquelitinib (0–40 nM). CD19 CAR-T cells were co-cultured with HBL-1 tumor cells and subjected to repetitive antigen stimulation over a 7-day period; n = 3. I, J Summary of percentages of 2B4⁺, CD39⁺ and CD38⁺ cells within CD4⁺ and CD8⁺ CD19 CAR-T cell populations; n = 4–5. K, L The expression of nuclear transcription factors BATF was analyzed within CD4⁺ and CD8⁺ CAR-T cells by flow cytometry; n = 5. M The expression of p-PLCγ-1and PLCγ-1 were analyzed by western blotting (GAPDH used as loading control). N The expression of transcription factors TCF1, NR4A, NFAT1, TOX and BATF were analyzed by western blotting (GAPDH used as loading control). O The expression of IFN-γ, TNF-α and LAG3 were analyzed by western blotting (GAPDH used as loading control).