Fig 9. DC subtypes, CD8 cytotoxic degranulation, and CD8+CD122+ Treg cells in splenocytes of immunized mice.
Splenocytes from the different experimental groups were treated with 20μg/ml tCRAMP for 24 hours and stained for markers of DC subtypes (A-D). Gating is as depicted in S5 Fig. Bars over graph columns indicate statistical significance; P<0.05. Mean of 3–5 separate experiments with at least 2 mice pooled per group in 3–4 replicates per experiment. Splenocytes from mice immunized with 20μg Cramp, adjuvant, or PBS were treated with 20μg/ml tCRAMP for 4 hours in the presence of 2μg/ml fluorescently labeled CD107a monoclonal antibody. Cells were also stained for CD8b and CD122, and analyzed for CD107a+ degranulation (E) or CD8+CD122+ Treg cells (F). Analysis excluded CD49b+ cells. Isotype staining was used as control. Results of the analysis were plotted on bar graphs. Bars over graph columns indicate statistical significance; P<0.05.