Fig. 4: CLDN18.2 and mesothelin AND-gate mitigates CAR-T cell toxicity.
From: On-target off-tumor toxicity of claudin18.2-directed CAR-T cells in preclinical models
A Schematic of the LINK-based AND-gate strategy. CAR-T cell activation and killing occur only upon simultaneous recognition of both Claudin18.2 and Mesothelin on target tumor cells. B In vitro testing of the AND-gate in Jurkat NFAT-GFP reporter cells. Jurkat cells expressing one or both receptors were cultured alone or co-cultured with wild-type (wt), Mesothelin knockout (MesothelinKO), or Claudin18.2 knockout (Claudin18.2KO) GSU cells. After 16 h, cells were analyzed for GFP expression via flow cytometry. C Real-time cytotoxicity assay comparing UTDs, hu8e5-CD28z, and LINK CAR-T cells in co-culture at a 3:1 E:T ratio with the gastric cancer cell line GSU (MesoKO, Claudin18.2KO or wt) (n = 2 healthy donors. Average of three independent replicates, mean ± s.d., 2-way ANOVA). D, E In vivo comparison hu8e5-CD28z and LINK CAR-T cells (n = 2 healthy donors, 5 mice per group/donor). NSG mice were injected subcutaneously with 2 × 10⁶ GSU tumor cells. After 10 days, mice were treated intravenously with 3 × 10⁶ CAR-T cells D Body weight change compared to pre-treatment (mean ± s.d., 2-way ANOVA). E Tumor volume (mean ± s.d., 2-way ANOVA).
