Fig. 1 | Signal Transduction and Targeted Therapy

Fig. 1

From: Dual epitope anti-LILRB4 synthetic T-cell receptor and antigen receptor (STAR)-T-cell therapy for relapsed/refractory acute myeloid leukemia

Fig. 1The alternative text for this image may have been generated using AI.

DE STAR-T cells exhibited potent cytotoxicity against AML cells in preclinical assays. a Top: graphic NLB4-STAR, NLB14-STAR and DE-STAR structure; bottom: Gene schematics of anti-LILRB4-STAR in lentiviral vectors fused with a red fluorescence protein reporter. b, c DE STAR-T cells exhibited more vigorous cytotoxicity than monovalent STAR-T cells against MV4-11 or KASUMI-1 cells at a low E:T ratio. The results are presented as the mean ± SEM, and p values were determined by two-way ANOVA. ***p < 0.001. d DE STAR-T cells produced more cytokines than NLB-4 or NLB-14 STAR-T cells did after 24 h of coculture. The E:T ratio was 1:1, and ELISA was used to determine the IFN-γ, IL-2, and TNF-α concentrations in the medium. The results are presented as the mean ± SEM, and p values were determined by one-way ANOVA. ***p < 0.001, ##p < 0.01. e, f APOE and FN did not affect DE STAR-T cells conquering the target cell. The E:T ratio was 1:1, and the target cell line was the MV4-11 cell line. The results are presented as the mean ± SEM, and p values were determined by one-way ANOVA. ns means not significant. g The cytotoxicity of the DE STAR-T cells was stronger when the cells were cocultured for 24 h. The results are presented as the mean ± SEM, and p values were determined by one-way ANOVA. ***p < 0.001. h The percentage of LILRB4-positive cells among different cell types in patient-derived bone marrow. i Changes in the morphology of MV4-11 cells and CD34+ HSCs cocultured with DE STAR-T cells. The MV4-11 cell line group served as the positive control. j Most CD34+ HSC survival after coculture with DE STAR-T cells for 24 h. K CD34+ HSCs induced activation in only a small subset of DE STAR-T cells after coculture for 24 h. l Schematic of the monitoring of tumor elimination by monovalent or bivalent LILRB4-STAR-T cells in the MV4-11-luc xenograft tumor model. m Bioluminescence images of MV4-11-luc cell inhibition in each group monitored by Lumina II (CALIPER) after intraperitoneal D-luciferin (YEASEN) injection. n Quantification of fluorescence intensity in each group. The results are presented as the mean ± SD, and p values were determined by two-way ANOVA. *p < 0.05, ***p < 0.001. o Persistence and proliferative capability of monovalent or bivalent STAR-T cells measured by the number of RFP+ cells in mouse peripheral blood samples. The results are presented as the mean ± SD, and p values were determined by one-way ANOVA; */#p < 0.05, **/##p < 0.01, ***p < 0.001, ns indicates not significant. p LILRB4, CD123 and CLL1 expression levels in patient-derived tumor cells. The results are presented as the mean ± SEM, and p values were determined by one-way ANOVA. ***p < 0.001. q Graphic of the DE-STAR and DE-CAR structures. r Compared with different DE-CAR-T cells, DE-STAR-T cells exhibited more vigorous cytotoxicity against the KASUMI-1 cell line at various E:T ratios. The results are presented as the mean ± SEM, and p values were determined by two-way ANOVA. ***p < 0.001. s Compared with different DE-CAR-T cells, DE-STAR-T cells produced more cytokines against the KASUMI-1 cell line after 24 h of coculture. The E:T ratio was 1:2, and ELISA was used to determine the IFN-γ, IL-2, and TNF-α concentrations in the medium. The results are presented as the mean ± SEM, and p values were determined by one-way ANOVA. ***p < 0.001, ##p < 0.01. t DE STAR-T cells exhibited robust TCR and ncNFκB signaling pathway activation after KASUMI-1 cells were stimulated for different durations. u Schematic of the monitoring of tumor elimination by DE STAR-T cells and DE-CAR-T cells in the MV4-11-luc xenograft tumor model. v Quantification of fluorescence intensity in each mouse. w Proliferative ability of DE STAR-T cells and DE-CAR-T cells, as measured by the number of RFP+ cells in mouse peripheral blood samples. The results are presented as the mean ± SD, and p values were determined by one-way ANOVA; *p < 0.05; ns not significant

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