Journal: Cancer research

Article Title: Armored Chimeric Antigen Receptor-T Cell Therapy Targets Antigen-Heterogeneous Glioma

doi: 10.1158/0008-5472.CAN-26-1515

Figure Lengend Snippet: (A–B) C57BL/6J mice were implanted intracranially with a mixture of 5e4 muIL-13Rα2+ CT-2A cells and 5e4 muIL-13Rα2− CT-2A cells. Six days later, mice were injected intratumorally with 5e5 CAR+ murine T cells per mouse. N = 7 per group. (A) Timeline of anti–TNF-α antibody injection. (B) Tumor growth as monitored by bioluminescence imaging. (C) CD90.2+ C57BL/6J mice were injected with tumor and CD90.1+ T cells as described in Fig. 2A and sacrificed two or nine days post ACT. N = 3 per group. CD90.2+ cells isolated from each brain were co-cultured with IL-13Rα2− CT-2A cells at a 1:1 effector-to-target ratio for 44 hours. Activation was assessed by cytokine release using FluoroSpot plate. The assay contained biological triplicates (i.e., 3 mice) per treatment group, with each biological replicate seeded in technical duplicates (i.e., 2 wells per mouse; 6 wells per treatment group). Each dot in the plot represents 1 mouse, with a value equal to the mean number of spots between technical duplicate wells. ****, P < 0.0001; ***, P < 0.001; **, P < 0.01; *, P < 0.05; ns, not significant. (D) C57BL/6J mice were implanted intracranially with a mixture of 5e3 muIL-13Rα2+ SB28-GKO cells and 5e3 muIL-13Rα2− SB28-GKO cells. Six days later, mice were injected intratumorally with 5e5 CAR+ murine T cells per mouse. Tumor growth as monitored by bioluminescence imaging. N = 10 per group.

Article Snippet: Subsequently, red blood cells were lysed using Miltenyi 1x Red Blood Cell Lysis solution, and T cells were enriched with a REAlease ® CD90.2 MicroBead Kit (Miltenyi, Cat# 130-126-706).

Techniques: Injection, Imaging, Isolation, Cell Culture, Activation Assay