Figure 2.
Viral Antigen Stimulation Significantly Increases CAR Expression in PB-Modified CD19-Specific T Cells
(A) Schematic explanation of the generation method of PB-modified CD19.CAR-T cells in the CD3/CD28-ATC or the ACE condition. 1 × 107 of PBMCs were transfected with CD19.CAR via the piggyBac transposon system and stimulated with either irradiated autologous ATCs and CD3/CD28 mAbs (CD3/CD28-ATC CAR method) or irradiated autologous ATCs pulsed with viral peptides (ACE CAR method) on days 0 and 7 after nucleofection. The cells were harvested and analyzed on day 14. (B) Ex vivo expansion of PB-modified CD19.CAR-T cells. Cell numbers were determined by a trypan blue exclusion assay on days 7 and 14. Data are presented as the mean ± SD of experiments from nine donors. (C and D) CD19.CAR expression on PB-modified T cells was examined by flow cytometry. Representative (C) and summary (D) results from nine donors are shown. The results are presented as the mean ± SD. (E) CAR+ T cell numbers are calculated based on the total cell numbers and CD3+ CAR+ percentages determined by flow cytometry. Data are presented as the mean ± SD from nine donors.