Journal: bioRxiv
Article Title: Multicenter preclinical validation of next-generation CAR T cells: a strategy for harmonization, reproducibility, and its feasibility in clinical translation
doi: 10.64898/2026.04.10.717659
Figure Lengend Snippet: CAR T cell effector functions were assessed by ELISA recording IFN-ƴ, XTT-based viability assay, and transwell migration assay across two independent centers. (A–C) IFN-γ secretion measured after T cell coculture with Panc02-EpCAM cells (A, 24 h, 10:1 E:T ratio), SUIT-mesothelin cells (B, 48 h, 5:1 E:T ratio), or LS174T cells (C, 24 h, 5:1 E:T ratio); supernatants were diluted to 1:500 (EpCAM and CEA) or 1:200/1:1,000 (mesothelin, centers A/B). (D–F) Cytotoxicity was assessed after 48 h coculture with Panc02-EpCAM (D, 10:1 E:T), 48 h with SUIT-mesothelin (E, 5:1 E:T), or 24 h with LS174T (F, 5:1 E:T). (G–I) In vitro migration was recorded in a transwell system toward 2 ng/ml murine CCL1 (G, EpCAM) or 10 ng/ml human CCL1 (H, mesothelin; I, anti-CEA). Gray bars depict manipulation checks (A–F, CAR > GFP; G–I, odds ratio = 1 ± 0.25 for GFP and CAR), and black bars confirmatory test results (A–C, CCR8-CAR > GFP; D–F, CCR8-CAR > GFP and CCR8-CAR > CAR; G–I, primary: CCR8-CAR > CAR, secondary: CCR8-CAR > GFP). Mean ± standard deviation was calculated across both centers. Y-axis scaling differs between murine (A, G) and human models (B–C, H–I). Statistical details reported in main text.
Article Snippet: Murine Panc02-EpCAM and human SUIT-mesothelin target cell lines have previously been engineered to express the respective target antigen ( ); the LS174T (ATCC-CL188, RRID:CVCL_1384) cell line endogenously expresses CEA.
Techniques: Enzyme-linked Immunosorbent Assay, Viability Assay, Transwell Migration Assay, In Vitro, Migration, Standard Deviation
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