Fig. 1: On-target Off-tumor toxicity of CLDN18.2 CAR-T cells.
From: On-target off-tumor toxicity of claudin18.2-directed CAR-T cells in preclinical models
A hu8e5-2I ScFv (hu8e5) and Zolbetuximab ScFv (Zolbe) containing anti-CLDN18.2 CAR T cell design. B Real-time cytotoxicity assay of CLDN18.2 CAR T co-culture at a 1:1 E:T ratio with the gastric cancer cell lines NUGC-4 (left) or GSU (right) (mean of three independent replicates, mean ± s.d., 2-way ANOVA). C Epitope mapping of Zolbe and hu8e5 CARs. Zolbe-CD28z or hu8e5-CD28z CAR-expressing NFAT-GFP Jurkat cells were cultured for 16 h with wt Jurkat cells (WT), or Jurkat cells expressing human Claudin 18.1, Claudin18.2, or Claudin18.2 variants and analyzed by flow (normalized GFP expression compared to Claudin18.2, median of 6 technical replicates, Kruskal–Wallis test). D Experimental schematic (created in BioRender. Birocchi, F. (2025) https://BioRender.com/foaa417). NSG mice were injected subcutaneously with 2 × 106 GSU tumor cells. After 10 days mice were treated with 3 × 106 CAR-T cells (intravenously). E Flow cytometry plot showing CAR-T cell transduction efficiency. Tumor volume (F) and body weight change compare to pre-treatement (G) of GSU-bearing mice untreated (Tumor only), or treated with untransduced T cells (UTDs), Zolbe-CD28z, or hu8e5-CD28z (mean ± s.d., 2-way ANOVA) (n = 5/6 mice per group). H Absolute CAR-T cell count in the peripheral blood of treated mice at the end of the experiment (day 22) (n = 3 mice per group). I Immunohistochemistry staining of representative mice from the same experiment of D-F showing Claudin18.2 expression and T cell infiltration (hCD3) in CLDN18.2 CAR-T treated mice stomach compared to UTDs. FS = forestomach, GS = glandular stomach, LR = limiting ridge.
