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. Author manuscript; available in PMC: 2025 Jan 1.
Published in final edited form as: Methods Mol Biol. 2024;2748:201–241. doi: 10.1007/978-1-0716-3593-3_15

Figure 6. CAR-mediated activation of human CAR+ T cells.

Figure 6.

A. The workflow for evaluating CAR-mediated early activation of CAR T cells, e.g. CD19CAR Tregs, involves co-incubating CD19CAR Tregs with irradiated CD19-expressing K562 for 48h, followed by flow cytometry to detect upregulation of the activation marker CD71 (transferrin receptor). Continued expansion of the cells for 9 days allows for assessment of CAR-mediated expansion (CAR Treg cell number), as well as assess the phenotypic stability of the CAR Tregs (e.g. FOXP3 expression). B. CD19CAR-mediated activation of human CD4+ T cells, CD8+ T cells, and Treg cells after 48h co-incubation with either irradiated K562 or CD199-K562, as assessed by flow cytometry with anti-human CD71 FITC. Histograms represent CD71 surface expression of live CD4+GFP+ cells.