Figure 3.

In Vitro Bioluminescence Imaging and [¹⁸F]-TMP Uptake of DYR-Transduced Primary Human T Cells

(A) Primary T cells were transduced with pELPS DHFR-YFP-Renilla (DYR) and sorted on YFP expression. 1 × 10⁵ DYR T cells or NTD T cells were incubated with 1.5 μM coelenterazine in a 96-well plate and subjected to bioluminescence imaging (BLI). Mean total flux is shown for each group (n = 3). (B) 1 × 10⁶ DYR or NTD T cells were incubated with 2 × 10⁶ cpm of [¹⁸F]-TMP for 30 min prior to washing with PBS. Uptake as a %ID was measured by gamma counting and normalized per million cells (n = 3). (C) Primary human T cells that had been activated with anti-CD3/CD28 antibody-coated beads were co-transduced with DYR lentivirus as well as lentivirus encoding the GD2-E101K-4-1BB CAR containing an mCherry fluorescent protein separated by a T2A site. The population of double-positive (DP) T cells was isolated by flow cytometric cell sorting. (The collected DP population is shown in red.) T cells within the YFP gate (blue) were collected to serve as CAR-negative control DYR T cells. Transduction efficiencies for the lentiviral vectors are shown in the inset. Error bars represent the SEM.