Fig. 2.
CD8-dependence of the generated CTLs. C57-A2Kb transgenic mice (3 mice) were immunized with 3 × 106 syngenic DCs pulsed with mp30 (10 μg/ml) and PADRE (5 μg/ml). Ten days after immunization, the mice were sacrificed and the pooled splenocytes (from 3 mice) were pulsed with mp30 (100 μg/ml) and expanded in vitro for 5 days. Day 5, splenocytes (300,000/well) were tested in ELISPOT wells for IFN-γ production when stimulated with either EL4-A2 cells (100,000) pulsed with mp30 or an HLA-A*0201-binding irrelevant peptide (Mock) or EL4-A2Kb cells (100,000) pulsed with mp30 or an HLA-A*0201-binding irrelevant peptide (a). In some cases, splenocytes were tested in the 4 h cytotoxicity assay (b). Target cells were mp30-pulsed EL4-A2Kb and incubated with effector cells at ratio of 1:50 in the presence of anti-CD8 antibody or isotype control. *P < 0.001 (t test)