Fig. 3

CLL-1 CAR-T cells lyse CLL1-expressing tumor cells. a Expression of CLL-1 on the cell line Raji-CLL1, Raji, U937, and K562. b CLL-1 CAR-T cells lysed CLL-1⁺ cell lines Raji-CLL1 and U937. CLL-1⁻ cell lines Raji and K562 were used as negative control. NT cells were used to evaluate unspecific lysis. Data represent mean values of triplicate wells ± SD. c CLL-1 CAR-T cells underwent proliferation to CLL-1 antigen. CFSE-labeled target cells were co-cultured with CAR-T cells for 4 days in the absence of exogenous cytokines at an E:T ratio of 1:1. Cells were stained using anti-CD3 to distinguish between T cells and CFSE-labeled tumor cells. Percentages in each quadrant are indicated. d Expression of CLL-1 on three primary AML samples used in the in vitro killing assay. e CLL-1 CAR-T cells displayed cytotoxicity to autologous primary AML cells. CLL-1 CAR was transduced into T cells derived from the patients and cultured with autologous CD34-enriched AML cells for 24 h at the indicated E:T ratios; NT cells were used as negative controls. Data represent mean values of triplicate wells ± SD. f Cytokine profiling of CLL-1 CAR-T or NT cells in response to a 24-h co-culture with various target cells. Data represent mean values of triplicate wells ± SD. GM-CSF granulocyte macrophage colony-stimulating factor, IFN-γ interferon-γ, IL-13 interleukin-13, IL-2 interleukin-2, MIP-1α macrophage inflammatory protein-1α, TNF-α tumor necrosis factor α