In vitro and in vivo model of a novel immunotherapy approach for chronic lymphocytic leukemia by anti-CD23 chimeric antigen receptor

Supplemental materials for: Attianese et al

Files in this Data Supplement:

• Figure S1. Cytokine production and proliferation of CD23.CAR+ T cells in response to MEC-1 and autologous CLL cells (JPG, 79.9 KB) -
  CD23.CAR+ T cells produced significant amount of immunostimulatory cytokines as compared to NT cells when co-cultured with either MEC-1 IFN-γ (2063 pg/ml, range 1309�2574 pg/ml vs. 197 pg/ml, range 0�697 pg/ml, n=5, p≤0.005 ), TNF-α (693 pg/ml, range 115�1300 pg/ml vs. 5 pg/ml, range 0�21 pg/ml, n=5, p≤0.005 ) and TNF-β (484.3 pg/ml, range 73�882 pg/ml vs. 24 pg/ml, range 0�59 pg/ml n=5, p≤0.005) (A). CD23.CAR+ T cells also produced significant amount of immunostimulatory cytokines as compared to NT cells also when co-cultured with primary autologous non irradiated CLL cells in CLL patient 1 (IFN-γ average level of 1002 pg/ml, range 965�1039 pg/ml vs 0 pg/ml; TNF-α average level of 386 pg/ml, range 361�411 pg/ml vs 0 pg/ml; TNF-β average level of 61 pg/ml, range 60�62 pg/ml vs 0 pg/ml) (B), and CLL patient 2 (IFN-γ average level of 147 pg/ml, range 145�147 pg/ml vs 0 pg/ml; TNF-α average level of 23 pg/ml, range 11�34 pg/ml vs 0 pg/ml; TNF-β average level of 10 pg/ml, range 9�11 pg/ml vs 0 pg/ml) (C). CD23.CAR+ modified T cells proliferated in response to either MEC-1 cell line (E:T Ratio 1:1) (average fold increase of 50, range 26�83, n=5, p≤0.05) as compared to NT cells after three week of culture without the addition of exogenous cytokines (D) or in response to non-irradiated primary autologous CLL cells (E:T Ratio 1:1) (panels E for patient 1 and F for patient 2).