Figure 6.

Upregulation of CD123 expression after activation of CAR T cell. (A) The proliferation of CART123 and NT after CFSE-stained CART123 and NT were stimulated with IL-2 100 U/mL and CD3/CD28 beads at 1:1+ IL-2 100 U/mL for 48 hrs; then, CFSE dilution was analyzed by flow cytometry. (B) Expression of CD123 and CAR after stimulation for 48 hrs was analyzed by flow cytometry (mean±SEM, n=3). Data show one representative experiment. (C) The proliferation of CART123 and NT were stimulated with IL-2 and CD3/CD28+ IL-2, or treated with different concentrations of anti-CD123 antibodies for 48 hrs (mean± SEM, n=3). Data show one representative experiment. (D-E) CART123, CART19 or NT cells from healthy donors were activated by CD3/CD28 beads at 1:1 and cultured in vitro with IL-2 100 U/mL. Six days after transduction, T lymphocytes were analyzed by flow cytometry. (D) Representative plots are shown. (E) (mean±SEM, n=3). Data show one representative experiment. (F) Proliferation curve of CART123, CART19, and NT within 7 days after transfection (mean±SEM, n=3). Data are depicted from three independent experiments; each point represented an average of triplicate. (G) Apoptosis detected by 7-AAD and Annexin V at day 7 after transfection (mean±SEM, n=3). Data are depicted from three independent experiments; each point represented an average of triplicate. *P<0.05; **P<0.01; ***P<0.001; ****P<0.0001.

Abbreviations: CAR, chimeric antigen receptor; NT, non-transduced T; 7-AAD, 7-Aminoactinomycin D; CFSE, carboxyfluorescein succinimidyl ester.