Anti-CD123 (Klon43) h(41BB-CD3ζ) CAR, pCDCAR1 vector

Complete CAR data Cyt

Fig.1 Cytotoxicity of CD123 CART against CAL-1 BPDCN cells in vitro.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.1 Cytotoxicity of CD123 CART against CAL-1 BPDCN cells in vitro.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.2 Flow cytometry analysis of CD107α degranulation on CD8+ cells after co-culture with CD123 CART or TCRαβ KO T cells at E:T = 1:1 for 6 h.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.2 Flow cytometry analysis of CD107α degranulation on CD8+ cells after co-culture with CD123 CART or TCRαβ KO T cells at E:T = 1:1 for 6 h.

"T cells only" correspond to the basal activity of unstimulated T cells, while PMA/Ion corresponds to the signal observed upon nonspecific stimulation by phorbol myristate acetate and ionomycin.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.3 IFNγ levels determination after CAL-1 cells were co-cultured with either TCRαβ KO T cells or with CD123 CART cells at E:T = 1:1 for 25 h.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.3 IFNγ levels determination after CAL-1 cells were co-cultured with either TCRαβ KO T cells or with CD123 CART cells at E:T = 1:1 for 25 h.

IFNγ levels were determined by the BioLegend LEGENDplex assay.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.4 Antitumor activity of UCART123 against primary BPDCN samples in vitro.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.4 Antitumor activity of UCART123 against primary BPDCN samples in vitro.

Specific cytotoxic activity against target cells (CD123+ MOLM13 cells and primary BPDCN samples) upon co-culture for 16 h with either UCART123 cells or TCRαβ KO T cells at E:T = 10:1.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.5 Flow cytometry analysis of CD107α degranulation after co-culture with CD123 CART or TCRαβ KO T cells.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.5 Flow cytometry analysis of CD107α degranulation after co-culture with CD123 CART or TCRαβ KO T cells.

Degranulation activity upon co-culture of UCART123 or TCRαβ KO T cells with CD123- Jurkat cells as well as CD123+ MOLM13 cells and five different primary BPDCN samples is shown.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.6 IFNγ levels determination upon co-culture of UCART123 cells or TCRαβ KO T cells with CD123- or CD123+ cells at E:T = 1:1 for 25 h.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.6 IFNγ levels determination upon co-culture of UCART123 cells or TCRαβ KO T cells with CD123- or CD123+ cells at E:T = 1:1 for 25 h.

IFNγ levels were determined using the BioLegend LEGENDplex assay. PMA/ionomycin was used as a positive control.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.7 UCART123 treatment results in long-term survival of primary BPDCN PDX.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.7 UCART123 treatment results in long-term survival of primary BPDCN PDX.

a Experimental design. b Survival of the mice in indicated treatment groups was estimated by the Kaplan-Meier method. c Circulating tumor burden was measured by flow cytometry in peripheral mouse blood samples collected on the indicated days (post treatment).

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Complete CAR data FuncS

Fig.8 Loss of CD123 leads to escape from UCART123 therapy and causes early relapses.

CAR Construction : Klon43 scfv-41BB-CD3ζ;

Fig.8 Loss of CD123 leads to escape from UCART123 therapy and causes early relapses.

a Experimental design. b Survival of mice in the treatment groups was estimated by the Kaplan-Meier method. Red arrow indicates start of treatment (Day 19). c Expression of CD123 on tumor cells isolated from bone marrow of control or UCART123-treated mice at the time they were sacrificed due to excessive tumor burden.

Cai, T., Gouble, A., Black, K. L., Skwarska, A., Naqvi, A. S., Taylor, D., ... & Konopleva, M. (2022). Targeting CD123 in blastic plasmacytoid dendritic cell neoplasm using allogeneic anti-CD123 CAR T cells. Nature Communications, 13(1), 1-11.

Anti-CD123 (Klon43) h(41BB-CD3ζ) CAR, pCDCAR1 (CAR-LC175)

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