CD19-CAR-T cells with short and long spacers show specific in vitro function. CD19-CAR-T cells with short and long spacers show specific in vitro function.

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CD19-CAR-T cells with short and long spacers show specific in vitro function. CD19-CAR-T cells with short and long spacers show specific in vitro function. A, design of lentiviral transgene inserts encoding CD19-specific CARs with different extracellular spacer lengths. B, analysis of EGFRt expression on CD8+ TCM–derived T cells transduced with the lentiviral vectors encoding the CD19-CARs with short or long spacer domains before enrichment (pre) and after enrichment and expansion (post). Cytolitic activity (C), IFNγ production (D), and proliferation (E)of CD19-CAR-T cells after coculture with CD19+ (K562/CD19, Raji) and control (K562) target cells. Proliferation after stimulation with CD19− K562 cells is shown as a comparison in each histogram (gray). Numbers above each histogram indicate the number of cell divisions the proliferating subset underwent, and the fraction of T cells in each gate that underwent ≥4/3/2/1 cell divisions is provided in the upper left of each plot. Data in B to E are representative of experiments with CAR-T cells derived from at least three different donors. Michael Hudecek et al. Cancer Immunol Res 2015;3:125-135 ©2015 by American Association for Cancer Research