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. 2022 Sep 8;7(17):e160674. doi: 10.1172/jci.insight.160674

Figure 1. Generation of hCAR19 Tregs.

Figure 1

(A) Schema of hCAR19 and tEGFR Treg generation. (B) Schematic representation of hCAR19 construct in a pMP71 vector backbone. scFv, single chain variable fragment; VL, variable light chain; VH, variable heavy chain; TM, transmembrane domain; GMCSFRss, GM-CSF receptor α chain signal sequences. (C) Representative histogram plots of truncated human epidermal growth factor receptor (tEGFR) expression in hCAR19 and tEGFR control transduced Tregs prior to experimental use. FMO, fluorescence minus one; pre-E, before tEGFR-positive enrichment; post-E, following tEGFR-positive enrichment. (D) Percentage of tEGFR expression in nontransduced (NT) Tregs, before and after tEGFR enrichment of hCAR19 and tEGFR transduced Tregs. (E) Histogram plots of CD25 and Foxp3 expression in NT, tEGFR, and hCAR19 Tregs. (F) Representative FACS plot of CD25+Foxp3+ transduced hCAR19 Tregs prior to experimental use.