Figure 5.
Impaired apoptotic cell uptake and cross-presentation by CD8+ DCs in CD11c-Rac1(N17) mice. (A) Tg+ mice and WT controls were injected with 1 × 107 CFSE-labeled allogeneic (BALB/c) B cells, and spleens were harvested 14 hours later. Following collagenase/DNase digestion, splenocytes were stained with mAb specific for CD11c and CD8α. Dot plots are gated on CD11chi cells, with CFSE labeling and CD8 staining as indicated. The frequencies of CFSE+ cells among all DCs (CD11chi) and CD8+ DCs are shown (mean ± SD, n = 3). Data are representative of 2 independent experiments with similar outcome. (B) CFSE-labeled OT-1 cells (1 × 106) were transferred into Tg+ mice and WT controls, followed by injection of 1 × 106 OVA- or BSA (control)–loaded β2m-/- cells 1 day later. Spleens were harvested 3 days after the second injection, digested with collagenase/DNase, and stained with anti-CD8α mAb, H2-Kb/SIINFEKL tetramers, and PI. Dot plots are gated on CD8+ cells, with CFSE labeling and tetramer staining as indicated. Histogram analysis of CFSE+ OT-1 cells is shown as inserts in the dot plots. Total numbers of OT-1 (Kb/SIINFEKL tetramer-positive) cells per spleen (upper graph) and OT-1 cells per cell division (lower graph) were calculated; the mean (± SD) is shown for 3 mice per group (BSA, n = 2). Results from one experiment are shown and are representative 3 independent experiments performed.

Impaired apoptotic cell uptake and cross-presentation by CD8+ DCs in CD11c-Rac1(N17) mice. (A) Tg+ mice and WT controls were injected with 1 × 107 CFSE-labeled allogeneic (BALB/c) B cells, and spleens were harvested 14 hours later. Following collagenase/DNase digestion, splenocytes were stained with mAb specific for CD11c and CD8α. Dot plots are gated on CD11chi cells, with CFSE labeling and CD8 staining as indicated. The frequencies of CFSE+ cells among all DCs (CD11chi) and CD8+ DCs are shown (mean ± SD, n = 3). Data are representative of 2 independent experiments with similar outcome. (B) CFSE-labeled OT-1 cells (1 × 106) were transferred into Tg+ mice and WT controls, followed by injection of 1 × 106 OVA- or BSA (control)–loaded β2m-/- cells 1 day later. Spleens were harvested 3 days after the second injection, digested with collagenase/DNase, and stained with anti-CD8α mAb, H2-Kb/SIINFEKL tetramers, and PI. Dot plots are gated on CD8+ cells, with CFSE labeling and tetramer staining as indicated. Histogram analysis of CFSE+ OT-1 cells is shown as inserts in the dot plots. Total numbers of OT-1 (Kb/SIINFEKL tetramer-positive) cells per spleen (upper graph) and OT-1 cells per cell division (lower graph) were calculated; the mean (± SD) is shown for 3 mice per group (BSA, n = 2). Results from one experiment are shown and are representative 3 independent experiments performed.

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