Fig.2. scFv from
TR1419 CAR mediated tumor cell apoptosis by binding
with TRAIL-R1. (A) Schematic
representation of TR1419∆ζ and the lentiviral vector.
The construct consisted of the extracellular portion of anti-TRAIL-R1
scFv, linking via a CD8α hinge-transmembrane domains to the truncated
CD3ζ molecule. (B) The expression of TR1419-28BBζ and
TR1419∆ζ in Jurkat cells was detected by flow
cytometry. (C) Cytotoxicity of TR1419-28BBζ- and
TR1419∆ζ -Jurkat cells against SW480 in the absence or
in the presence of soluble TRAIL-R1 protein was determined by RTCA. The
analysis was performed using Students’ t tests. Data reflected the mean
± SEM of triplicate wells, ***p < 0.01. (D) SW480 were
fixed and labeled with annexin V and 7-AAD and then analyzed by flow
cytometry to identify apoptotic cells (annexin
V+/7-AAD− and annexin
V+/7-AAD+), after incubation with
effector cells for 2h. (E) The expression of caspase3 protein in SW480
analyzed by flow cytometry, after incubation with effector cells for 2h.