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Fig. 2 | Journal for ImmunoTherapy of Cancer

Fig. 2

From: A tandem CD19/CD20 CAR lentiviral vector drives on-target and off-target antigen modulation in leukemia cell lines

Fig. 2

Surface expression of single and tandem-CAR T constructs on human primary T cells. a CAR T expression was determined by flow cytometry. T cells were activated with Transact CD3 CD28 reagent in the presence of IL-2, and transduced with LV as described in Methods. On culture day 10, viable transduced T cells (7-AAD negative) were assayed for CAR surface expression using one of three staining reagents: Protein L (column 1), CD19 Fc followed by anti-Fc-AF647 (column 2), or CD20-biotin followed by streptavidin-PE staining (column 3). The LV used in transduction is listed to the left of each row. Percentage of CAR T-positive populations in relation to non-transduced T cell control is noted in the right-hand corner of each histogram. GFP-transduced cells served as an additional negative control. Representative data of three separate donors is shown. b The ratio of CD19 and CD20 antigen binding by each tandem-CAR is expressed as the ratio of percent cells bound by CD20 biotin vs CD19 Fc. The average + SD of three separate experiments using three donors is shown, **p < 0.01

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