Figure 2.
CAR-redirected T cells eliminate B-CLL cells predominantly via granule-mediated cytolysis. Anti-CD19scFv-CD3ζ CAR T cells were co-incubated (1 × 105 cells/well) with B-CLL cells (5 × 105 cells/well) in presence of the blocking anti-Fas-ligand antibody (10 μg/mL), the neutralizing anti-TNFα antibody (10 μg/mL), and EGTA (2 mM), respectively. Viability of B-CLL cells was monitored by flow cytometry after 18 hrs. As controls, the neutralizing capacities of the anti-TNF-α and anti-FasL antibodies were assessed by incubation of sensitive indicator cells with the respective reagents and antibodies in a cytotoxicity assay (data not shown).