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. Author manuscript; available in PMC: 2019 Jan 1.
Published in final edited form as: J Immunother. 2018 Jan;41(1):19–31. doi: 10.1097/CJI.0000000000000199

Figure 2. Absence of iC9 gene improves transduction efficiency and function of CD20 CAR T cells.

Figure 2

Healthy donor PBMC enriched for TCM were activated with anti-CD3/CD28 beads and transduced with a lentiviral vector containing either the iC9-1F5-NQ-28-BB-z construct or the same construct lacking the iC9 gene. Transgene expression was assessed by staining cells with anti-CD19 antibody to detect the tCD19 marker and analyzed by flow cytometry. (A) Histograms representing tCD19 expression of transduced cells (solid line) or untransduced control cells (filled histogram) are shown. The percent transduced cells is quantified in (B) and the geometric mean fluorescence intensity (MFI) of tCD19 expression is shown in (C). These results are representative of 3 independent experiments. (D) To measure cytokine secretion, TCM cells transduced with 1F5-NQ-28-BB-z or 1F5-NQ-28-z, with or without iC9 included, were co-incubated at a 1:1 ratio with irradiated Raji cells (left panel) or Jeko cells (right panel); supernatants were harvested 24 hours later and the indicated cytokines were measured by Luminex assay. IFN-γ and TNF-α are shown on the left y-axes and IL-2 is shown on the right y-axes. These results are representative of 2 independent experiments.