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Blood, Vol. 111, Issue 5, 2929-2940, March 1, 2008
Organ-derived dendritic cells have differential effects on alloreactive T cells
Blood Kim et al.
111: 2929
Supplemental materials for: Kim et al
Files in this Data Supplement:
- Figure S1. Unexpanded dendritic cells from MLN induce LPAM-1 (JPG, 27.8 KB)
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Unexpanded dendritic cells from PLN and MLN were cultured with naïve allogeneic T cells at a ratio of 1:2 in the presence of LPS (100 ng/ml). There is significantly higher expression of LPAM-1 in the MLN DC group as compared to the PLN DC group. (n=3, one of two experiments, *p=0.0012, p=0.0034 and p<0.0001, respectively).
- Figure S2. Kinetics of imprinting of homing molecules in vitro (JPG, 69.7 KB)
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Upregulation of homing molecules in vitro was examined after CFSE staining and analysed based on the number of cell divisions undergone. LPAM-1, E-lig and P-lig are rapidly upregulated on alloreactive T cells starting after the first cycle of cell division (one of three comparable experiments).
- Figure S3. Comparable secretion of IL12p40 by organ-derived dendritic cells (JPG, 22.4 KB)
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Determination of IL12p40 in cell culture supernatant shows comparable levels in all four groups (n=6).
- Figure S4. Comparable cytokine levels in sera of transplanted mice (JPG, 66.7 KB)
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Determination of ILp70, TNF , MCP-1 and IL10 levels in the sera of transplanted mice at day 14 showed no major differences (n=4-8).
- Figure S5. Unspecific stimulation of imprinted T cells leads to similar expression of activation markers (JPG, 76 KB)
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Imprinted T cells were additionally stimulated with PMA and ionomycin and their response was assessed by staining for intracellular IFN- and CD69. Expression of both markers is similar across all four groups. Of note, expression of IFN- seemed to be stronger in the LPAM-1+ subpopulations. CD4+ and CD8+ cells are gated on CD44hiCD62Llo cells.
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