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Blood, Vol. 112, Issue 7, 2878-2885, October 1, 2008
Dendritic cells mediate the induction of polyfunctional human IL17-producing cells (Th17-1 cells) enriched in the bone marrow of patients with myeloma
Blood Dhodapkar et al.
112: 2878
Supplemental materials for: Dhodapkar et al
Files in this Data Supplement:
- Figure S1. Frequency and profile of Th17 cells in healthy individuals (JPG, 72.4 KB)
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Peripheral blood mononuclear cells obtained from 20 healthy donors were stimulated with PMA and Ionomycin for 5 hours in the presence of golgistop and the frequency of IL17 producing CD3+ T cells was detected by intracellular cytokine staining. The cells were stained with aqua live/dead dye to gate out the dead cells, fixed, permeabilized and stained with antibodies against CD3, CD4, CD8, IL17, IL2 and IFNγ. (A) Figure shows % of CD3+ T cells that make IL17. The bar shows the mean 0.3 (SD=0.3 ). (B) Most of the IL17 producing T cells are CD4+. (C) Cytokine profile of the IL17 cells. We gated on the CD3+ IL17+ cells and asked if they were able to secrete cytokines in addition to IL17. Majority of the Th17 cells in healthy donors make either IL17 alone (mean 55% ± 17%) or IL17 with IL2 (mean 37% ± 15%). (D) Panel shows a representative FACS plot from one of the healthy donors. Most IL17+ T cells are CD4+. The plot on the right demonsrates the cytokine profile of the IL17 producing cells. Numbers in the quadrant represent % of IL17 cells secreting the cytokine.
- Figure S2. Expansion of Th17 cells from CD45RO depleted T cells (JPG, 56.8 KB)
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T cells were depleted of CD45RO+ populations using magnetic beads. Left panel shows the efficiency of depletion in the starting population. T cells were then cultured alone or with Cyt-DCs in the presence of 50 U/ml of IL2 on day 5. After another 7 days of co-culture, the cells were stimulated with PMA and ionomycin as previously and analyzed for the expansion of IL17 positive cells.
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