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Engineer and label cells
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Tumor cells are engineered to express one variant of luciferase (e.g., red-shifted).
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CAR T-cells are engineered to express a different luciferase variant (e.g., NIR-shifted).
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Establish mouse model
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Tumor cells are implanted into the mouse.
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CAR T-cells are introduced (e.g., by infusion).
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Administer Infraluciferin substrate
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Bioflares’ Infraluciferin methyl ester is delivered systemically.
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Once inside cells, esterases convert it into the active substrate.
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Capture dual bioluminescence signals
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Tumor cells and CAR T-cells emit distinct NIR signals.
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Imaging systems detect and quantify both signals non-invasively.
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Analyze therapeutic response
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Monitor CAR T-cell expansion, persistence, and trafficking.
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Track tumor regression (or relapse) in the same animal.
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Assess safety features such as CAR T-cell “suicide switches.”
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Advantages for CAR T-cell Studies
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Real-time efficacy assessment – Quantify tumor killing by CAR T-cells dynamically in living animals (Stowe et al., 2019).
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Superior accuracy – Far-red to NIR imaging penetrates deeper, improving quantification and resolution.
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Ethical and cost benefits – Dual imaging can halve animal usage by combining readouts into a single study.
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Safety validation – A powerful way to test built-in CAR safety mechanisms in vivo.
Summary
By combining Infraluciferin’s color-shifting properties with engineered luciferase variants (iLuc), dual NIR bioluminescence provides an unprecedented window into CAR T-cell therapy. Researchers can track tumor burden and immune activity together—non-invasively, longitudinally, and with high sensitivity—accelerating preclinical testing and improving translational outcomes.
