Figure 4
RNAi-mediated knockdown of ZAP70 affects T cell function in vitro and in vivo. DO11.10 T cells were transfected with 0.9 μM ZAP70-specific stealth™ siRNA #1 or as control with a control siRNA pool (Ctrl). 72 h after nucleofection, functionality of transfected cells was tested by restimulation (α-CD3) in vitro or by adoptive cell transfer into naïve recipient mice in vivo. (A) 1 × 106 DO11.10 T cells were restimulated with 3 or 10 μg/ml anti-mouse CD3 antibody. IL-2 production was determined in the supernatants by ELISA. Means of duplicate values are shown. (B) 1 × 106 DO11.10 T cells generated from DO11.10 mice were injected i.v. into Balb/c mice. 24 h after adoptive cell transfer, DTH response in the footpad was induced by s.c. injection of OVA peptide/IFA or PBS/IFA as control. Footpad thickness was determined 24 h after antigen exposure. (*** p = 0.001). Shown is the mean value of 20 mice in total (per group) analyzed in three independent experiments.