Fig. 1

NKG2D blockade during effector phase resulted in the formation of non-cytolytic memory CD8 T cells. a Schematic representation of the experimental design used to block NKG2D during the effector phase. At day 0, mice were immunized with peptide-loaded DC subcutaneously and injected retro-orbitally with purified pMel CD8 T cells. One week after immunization, half of the mice were injected intra-peritoneal with the anti-NKG2D blocking antibody (Ab) a day prior to the in vivo CTL assay. This period corresponds to the effector phase. Memory recall responses were analyzed at least one month later by repeating the in vivo killing assay. b Example of the in vivo killing assay readout by flow cytometry during memory responses. Immunized mice were injected with three populations of target splenocytes, each loaded with different amounts of CFSE and pulsed with different peptides. Spleens were analyzed 18 h later by flow cytometry and the ratios between the peptide-pulsed population vs. the unpulsed population were calculated and normalized to the naïve control mouse shown in the figure. The quantification of specific killing is summarized in the graph. Data shown is representative of four independent experiments