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Blood, Vol. 110, Issue 13, 4293-4302, December 15, 2007
Dendritic cells are specialized accessory cells along with TGF-β for the differentiation of Foxp3+ CD4+ regulatory T cells from peripheral Foxp3– precursors
Blood Yamazaki et al.
110: 4293
Supplemental materials for: Yamazaki et al
Files in this Data Supplement:
- Figure S1. DO11.10 RAG−/− mice lack Foxp3+ T regs (JPG, 85.3 KB)
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Spleen cells from DO11.10 RAG−/− or DO11.10 RAG+/+ mice were stained with anti-CD4 and CD25 mAbs, fixed, permeabilized, and stained with anti-Foxp3 or isotype control mAbs. Data are representative of 2 independent experiments.
- Figure S2. Antigen is necessary for the induction of Foxp3 (JPG, 120 KB)
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As in Fig.1A, but Foxp3−CD25−CD4+ T cells from DO11.10 RAG−/− mice were cultured with immature or mature BM-DCs (2 × 104) with or without TGF- in the presence or absence of OVA protein (500 µg/ml). Data are representative of 3 independent experiments.
- Figure S3. Effect of peptide dose on the induction of Foxp3 (JPG, 69.7 KB)
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(A) As in Fig.2, but Foxp3−CD25−CD4+ T cells (2 × 104) from DO11.10 RAG−/− mice were cultured with spleen CD11c+DCs (2 × 104) or CD11c−APCs (2 × 105) at the indicated doses of OVA peptide in the presence of TGF-. Cells were stained with mAbs to CD4, clonotype (KJ1.26), CD25, and Foxp3 or isotype control Abs. Representative gates of CD4+ KJ1.26+ cells are shown on the left. (B) As in A, but the absolute numbers of Foxp3+CD4+KJ1.26 clonotype+ T cells per culture are shown. Data are representative of 2 independent experiments.
- Figure S4. Foxp3+CD25+CD4+ T cells are induced from Foxp3−CD25−CD4+ T cells more effectively by spleen CD11c+ DCs than CD11c− APCs (JPG, 86.1 KB)
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The same experiment as Fig. 2 B,C was repeated. Foxp3−CD25−CD4+ T cells (2 × 104) from DO11.10 RAG−/− mice were cultured for 5 d with CD11c+ DCs (2 × 104) or CD11c− cells (2 × 105) from spleen and indicated doses of OVA peptide in the presence or absence of TGF- (2 ng/ml) and/ or IL-2 (100 U/ml). The absolute numbers of Foxp3+CD4+KJ1.26 clonotype+ T cells (top) and CD4+KJ1.26 clonotype+ T cells (bottom) per culture at 5 d are shown.
- Figure S5. Cytokine concentration in the culture supernatants from DCs plus TGF-β (JPG, 59.7 KB)
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As in Fig.3B, but Foxp3−CD25−CD4+ T cells (2 × 104) from DO11.10 RAG−/− mice were cultured with immature or mature BM-DCs (2 × 104) with indicated dose of peptide in the presence or absence of TGF-. After day 5, culture supernatants were collected, and the concentration of indicated cytokine was measured by Luminex.
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