Fig. 4
MHC IIhi migratory DCs are required for archived antigen presentation. a Schematic representation of experimental design. WT of BatF3 −/− mice were immunized subcutaneously as above 2 weeks prior to killing. Then lymph nodes were digested, T and B cells depleted, and then sorted on CD11c+ MHC class II high or intermediate expressing DCs. Approximately 20,000 DCs were co-cultured with 50,000 violet proliferation dye (VPD)-labeled OT1s for 3 days. b Flow cytometry of cells from a. Cells were gated on CD8+, CD45.1+ to evaluate activation (CD44) and division (VPD). c Quantification of samples from B where + is enriched lymph node DCs from a mouse immunized 1day prior and – is splenocytes from the BatF3 −/− mouse. Experiment was repeated twice. Six mice per group were killed and DCs sorted based on the gating strategy in a. Each point represents one well with 20,000 DCs and 50,000 VPD-labeled cells (based on the number of DCs recovered). Statistical analysis was done using an unpaired t-test where p = 0.0056 for WT and p = 0.0040 for Batf3 −/−. d Lymph node cells from mice above were stained for flow cytometry to evaluate the frequency of DCs that were MHC class II high between groups and numbers were calculated. An unpaired t-test was used to calculate the p-value of <0.0001
