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Firefly luciferase-based chronological measurement of effector CD8+ T-cell activity using a multi-chamber luminometer.

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Background: Although cell-mediated cytotoxicity has been evaluated with various protocols, methods for monitoring cytotoxicity in a time series have not been established. This work describes a method for evaluating cytotoxicity… Click to show full abstract

Background: Although cell-mediated cytotoxicity has been evaluated with various protocols, methods for monitoring cytotoxicity in a time series have not been established. This work describes a method for evaluating cytotoxicity using a multi-chamber real-time luminometer. Materials & methods: The efficiency of effector CD8+ T-cell expansion from melanoma-bearing splenocytes was analyzed. The effect of CD8+ T cells on the viability of luciferase-expressing target cells was measured by bioluminescence. Results: Melanoma-specific effector CD8+ T cells were differentiated by in vitro coculture. The melanoma cell growth was significantly inhibited in the presence of in vitro-expanded T cells in the bioluminescence-based time-lapse analysis. Conclusion: The bioluminescence-based assay is a useful method for monitoring the time course of cell viability of target tumor cells.

Keywords: cell; using multi; multi chamber; effector cd8; cd8

Journal Title: Bioanalysis
Year Published: 2023

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