Figure 4
Figure 4. CD103+ Treg cells induced apoptosis of autoantibody-secreting plasma cells. (A) Twenty days after infusion of CD103+ Treg cells, the spleen cells of the untreated and treated recipients were stained for B220 versus syndecan. The percentage of syndecan+ plasma cells and syndecan−B220+ B cells is shown beside the gating box. One representative of 4 examined recipients is shown. The means (± SE) of syndecan+ cells versus syndecan−B220+ cells in the untreated and treated recipients are 4.1% (± 0.4%) versus 1.0% (± 0.2%) and 34.5% (± 4.5%) versus 33.9% (± 3.8%), respectively. (B) Spleen cells (106) of the untreated recipients, including whole spleen (SPL), syndecan+ cell–depleted spleen (Syn−-SPL), sorted syndecan+ cells (Syn+), sorted syndecan−B220+ cells (Syn−), were cocultured with or without sorted CD103+ or CD103−CD4+ T cells (0.2 × 106) from the spleen of chronic GVHD recipients for 5 days. Anti-dsDNA IgG in the culture supernatant was measure by enzyme-linked immunosorbent assay. The mean (±SE) of 4 replicated experiments is shown. N/A indicates an antibody concentration below detectable levels. (C) Sorted activated syndecan+ and syndecan−B220+ B cells from chronic GVHD recipients and B220+ B cells from normal donors were cocultured with CD103+ or CD103−CD4+ T cells from chronic GVHD recipients for 8 hours. Thereafter, the cells were stained for annexin V versus DAPI. The percentage of annexin V+DAPI+ cells among Syn+ or sydecan−B220+ cells is shown beside the gating box. One representative of 4 replicated experiments is shown. The means (± SE) of the percentage of annexin V+DAPI+ cells among recipient syndecan+ cells are 5.4% (± 1.3%), 28.6% (± 3.6%), and 6.1% (± 1.7%), respectively, when cocultured alone or cocultured with CD103+ or CD103−CD4+ T cells. Similarly, the means (± SE) percentage among recipient syndecan−B220+ cells are 6.5% (± 1.9%), 15.4% (± 2.5%), and 6.8% (± 1.3%), and the mean (± SE) percentage among donor B220+ B cells are 5.5% (± 1.3%), 6.3% (± 1.5%), and 7.7% (± 1.4%).

CD103+ Treg cells induced apoptosis of autoantibody-secreting plasma cells. (A) Twenty days after infusion of CD103+ Treg cells, the spleen cells of the untreated and treated recipients were stained for B220 versus syndecan. The percentage of syndecan+ plasma cells and syndecanB220+ B cells is shown beside the gating box. One representative of 4 examined recipients is shown. The means (± SE) of syndecan+ cells versus syndecanB220+ cells in the untreated and treated recipients are 4.1% (± 0.4%) versus 1.0% (± 0.2%) and 34.5% (± 4.5%) versus 33.9% (± 3.8%), respectively. (B) Spleen cells (106) of the untreated recipients, including whole spleen (SPL), syndecan+ cell–depleted spleen (Syn-SPL), sorted syndecan+ cells (Syn+), sorted syndecanB220+ cells (Syn), were cocultured with or without sorted CD103+ or CD103CD4+ T cells (0.2 × 106) from the spleen of chronic GVHD recipients for 5 days. Anti-dsDNA IgG in the culture supernatant was measure by enzyme-linked immunosorbent assay. The mean (±SE) of 4 replicated experiments is shown. N/A indicates an antibody concentration below detectable levels. (C) Sorted activated syndecan+ and syndecanB220+ B cells from chronic GVHD recipients and B220+ B cells from normal donors were cocultured with CD103+ or CD103CD4+ T cells from chronic GVHD recipients for 8 hours. Thereafter, the cells were stained for annexin V versus DAPI. The percentage of annexin V+DAPI+ cells among Syn+ or sydecanB220+ cells is shown beside the gating box. One representative of 4 replicated experiments is shown. The means (± SE) of the percentage of annexin V+DAPI+ cells among recipient syndecan+ cells are 5.4% (± 1.3%), 28.6% (± 3.6%), and 6.1% (± 1.7%), respectively, when cocultured alone or cocultured with CD103+ or CD103CD4+ T cells. Similarly, the means (± SE) percentage among recipient syndecanB220+ cells are 6.5% (± 1.9%), 15.4% (± 2.5%), and 6.8% (± 1.3%), and the mean (± SE) percentage among donor B220+ B cells are 5.5% (± 1.3%), 6.3% (± 1.5%), and 7.7% (± 1.4%).

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