Supplementary Figure Legends, Figures 1 - 4 from Foxp3+ T Cells Inhibit Antitumor Immune Memory Modulated by mTOR Inhibition
PDF file - 308KB, Supplemental Figure 1. Tumor increases Tregs. The percent of CD4 cells expressing FoxP3 were examined from lymph nodes, spleen, bone marrow and peripheral blood lymphocytes (PBL) from tumor (~1 cm in diameter)bearing mice and control mice without tumor (n=5 per group). Flow cytometry results and summary data are shown. Histograms provide mean + SEM. *p<0.0,***p<0.005. Supplemental Figure 2. αCD4 antibody depletes effector CD4 subtypes. a. Splenocytes (n=3 per group) were collected before (day 0) and 1 or 10 days after administering αCD4 antibody, and analyzed by flow cytometry. The number of total splenocytes staining for CD4 and IL-2, IL-4 or IL-17 is shown. b. Percent of CD4 cells staining for IL-2, IL-4 or IL-17 is shown. Histograms provide mean + SEM. Supplemental Figure 3. CD4 depletion and mTor inhibition enhanced the antitumor effect of DC vaccine in B6 mice implanted with B16 tumor cells. B16 tumor cells were implanted into B6 mice (n=5 per group) on day 0. Tumor-lysate pulsed DCs were administered on day 3. CD4 lymphocytes were depleted with αCD4 antibody on days 6 and 9. Mice were treated with daily temsirolimus on days 9 to 23. Tumor growth was monitored. *p<0.05, **p<0.01,***p<0.005. Supplemental Figure 4. CD4 depletion prior to immune stimulation prevents formation of CD8 memory cells. The experimental scheme is similar to Figure 4 (n=5 per group). An additional group is included where αCD4 antibody is administered on day -1 and +1 (Group A). a. The percent of CD8 cells expressing Thy1.1 is shown from splenocytes harvested 5 days following rechallenge of memory cells. b. The percent of CD8 cells expressing Eomes is shown. Histograms provide mean + SEM. *p<0.05, **p<0.01,***p<0.005.