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. Author manuscript; available in PMC: 2015 Mar 16.
Published in final edited form as: J Immunol. 2012 Apr 4;188(10):4747–4758. doi: 10.4049/jimmunol.1103055

FIGURE 8.

FIGURE 8

Immune regulation by CD1d− B cells is not dependent on IL-10, but on cell–cell contact. (A) Purified BDC2.5 T cells were cocultured with the same number of sorted CD1d+CD5+ or CD1d−CD5+ B cells (all gated on CD19+B220+) from NOD mice in the presence of 10 ng/ml BDC2.5 mimotope peptide and irradiated APC (NOD splenocytes). Stimulation index (SI) was calculated as described earlier. Background cpm in the absence of mimotope was 74 ± 11. The experiments were performed twice, and similar results were obtained. (B) IL-10 production from the culture supernatants of (A) was examined by IL-10 ELISA kit (R&D Systems) following the manufacturer’s instructions. (C) Purified BDC2.5 T cells were cocultured with the same number of sorted CD1d+CD5+ or CD1d−CD5+ B cells (all gated on CD19+B220+) from IL-10−/− NOD mice in the presence of 10 ng/ml BDC2.5 mimotope peptide and irradiated APC (NOD splenocytes). Stimulation index was calculated as described earlier. Background cpm in the absence of mimotope was 128 ± 26. The experiments were repeated twice, and similar results were obtained. (D) Purified BDC2.5 T cells were cocultured with the same number of sorted CD1d+CD5+ or CD1d−CD5+ B cells (all gated on CD19+B220+), in 0.4-μm Transwells (BD Biosciences), from NOD mice in the presence of 10 ng/ml mimotope and irradiated APC (NOD splenocytes). Stimulation index was calculated as described earlier. Background cpm in the absence of mimotope was 52 ± 9. The experiments were performed twice, and similar results were obtained.