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. 2017 Jul 3;25(10):2270–2279. doi: 10.1016/j.ymthe.2017.06.026

Figure 5.

Figure 5

Detection, Isolation, and Elimination of Anti-CD19 CAR-T Cells with FR806 Co-expression

(A) Diagram of the lentiviral vectors used for the expression of FR806, anti-CD19 CAR, and GFP separated by F2A sequence. (B) Flow cytometry detection of FR806, anti-CD19 CAR, and GFP in FR806-CAR19-EGFP-transduced T cells. (C) 50:50 mixing assays were performed to test the depletion of FR806-CAR19-EGFP or mock-transduced T cells treated with 10 μg/mL CH12-MMAF. Survival of GFP+ cells was measured at the indicated time points. (D) T cells transduced with FR806-CAR19-EGFP (94.3% positive) could be isolated with biotinylated CH12 and anti-biotin microbeads. (E) Percentage of surviving UTD or sorted FR806-CAR19-EGFP+ T cells was measured by cell viability assay after incubation with the indicated concentrations of CH12-MMAF for 72 hr. Each of the data reflect the mean ± SD of triplicates from three different donors.