Skip to main content
. 2020 Jun 19;8(1):e000218. doi: 10.1136/jitc-2019-000218

Figure 5.

Figure 5

Venetoclax-resistant CLL cells can be killed using CD3xCD19 DART. (A) 3T3 or 3T40 treated CLL cells were cocultured for 24 hours with healthy donor T cells in the presence of 100 ng/mL CD3xCD19 dart and assessed for viability (n=10). (B) FACS analysis of lysis of WT Ramos or Ramos-Bcl-2 cells on coculture in 4:1 E:T ratio with HD T cells in presence of control or CD3xCD19 DART for 24 and 48 hours (n=4). (C) Specific lysis of coculture of primary CLL cells with untreated, DMSO treated or CMA treated HD T cells in a 4:1 ratio for 2 days (n=4). (D–E) Viability was assessed of mock transfected (n=1) and BAX/BAK KO clones (n=4) after treatment for 24 hours with (D) 10 μM venetoclax or (E) 100 ng/mL CD3xCD19 or the control dart molecule in 4:1 E:T ratio with HD T cells. (F–G) Specific lysis of primary CLL (F) treated with 100 nM ABT-199 for 24 hours in presence or absence of QVD-OPh (n=6) or (G) cocultured with HD T cells in 4:1 ratio and 100 ng/mL CD3xCD19 dart for 4 days in presence or absence of QVD-OPh (n=6). (H–I) Assessment of cleaved PARP by flow cytometry in (H) JeKo-1 cells cocultured for 24 hours or (I) CLL cells cocultured for 48 hours with HD T cells in a 4:1 ratio in the presence of 100 ng/mL CD3xCD19 DART, with or without 20 μM QVD-OPh (n=4–6). **P<0.01. ABT-199, Bcl-2 inhibitor venetoclax; CLL, chronic lymphocytic leukemia; CTV, cell trace violet; DART, dual affinity retargeting; HD, healthy donor; ns, not significant; WT, wild type.