Journal: Journal for immunotherapy of cancer
Article Title: Pooled screening for CAR function identifies novel IL-13Rα2-targeted CARs for treatment of glioblastoma.
doi: 10.1136/jitc-2024-009574
Figure Lengend Snippet: Figure 6 CAR 4 controls IL-13Rα2+ U87 cells and exhibits increased persistence in vitro. (A) Representative image of IL- 13Rα2+ U87-RFP tumor spheroids (magenta) in microdevices at day 3 and day 9 after CAR-T (cyan) addition. (B) Illustration of radial plot analysis carried out in (C) and (D). (C) Increase in U87 cell signal intensity at day 9 relative to day 0. (D) CAR-T signal at day 9. Data shown in (C,D) are means±SEM. (E) Illustration of regions inside and outside the microtumors. (F) CAR-T cell counts inside and outside the microtumors. (G) Activation, (H) exhaustion and (I) memory phenotypes of CAR-T cells retrieved from inside and outside the microtumors. P values in (C) were determined by comparing the area under the curve with one-way analysis of variance with Tukey’s multiple comparisons test and are <0.0001 (n=5 for untreated, 13BB𝜁 and CAR 4, df=12). P value in (D) were determined by comparing the area under the curve with two-way unpaired t-test and are <0.0001 (n=5 for 13BB𝜁 and CAR 4, df=4). Data in (F–I) are pooled from three technical replicates. CAR, chimeric antigen receptor; CM, central memory; EFF, effector; EM, effector memory; IFN, interferon; LAG-3, lymphocyte activation gene 3; MFI, mean fluorescence intensity; PD-1, programmed cell death protein 1; SCM, stem cell like memory; TIM3, T-cell immunoglobulin and mucin domain 3.
Article Snippet: U87 MG IL- 13Rα2+ RFP+ cells were generated by stable expression of human IL- 13Rα2 and RFP (Addgene plasmid #26001) and sorting for RFP and IL- 13Rα2 expression.
Techniques: In Vitro, Activation Assay, Fluorescence
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