In Vitro and In Vivo Antitumor Effect of Anti-CD33 Chimeric Receptor-Expressing EBV-CTL against Acute Myeloid Leukemia
EBV-CTLs could be stably transduced with the anti-CD33.CAR without alteration in their native immunophenotype and expansion rate. (a) The expression of the anti-CD33.CAR on the surface of EBV-CTLs was evaluated by flow cytometry with a Cy5-conjugated-mAb specific for the CH2CH3 domain of the CAR after 7 days of culture. A representative plot of EBV-CTLs transduction after 7 days of culture is shown. (b) The expression of CD4, CD8, CD3 along with CD56, CD4, and CD8 along with CD25, CD4, and CD8 along with CD45RA, CD4, and CD8 along with CD62L on the surface of EBV-CTLs was evaluated after 30 days of culture by flow cytometry. (c) Proliferation of anti-CD33.CAR-transduced EBV-CTLs compared to unmanipulated EBV-CTLs was evaluated by cell count with Trypan blue exclusion after weekly stimulations at 4 : 1 ratio with either irradiated autologous LCLs. Cells were cultured with low-dose rhIL-2 (20 U/mL). Data shown are mean ± SD of 6 separate experiments.
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