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Blood, Vol. 112, Issue 7, 2847-2857, October 1, 2008
Umbilical cord blood regulatory T-cell expansion and functional effects of tumor necrosis factor receptor family members OX40 and 4-1BB expressed on artificial antigen-presenting cells
Blood Hippen et al.
112: 2847
Supplemental materials for: Hippen et al
Files in this Data Supplement:
- Figure S1. Purification of Treg from frozen cord blood (JPG, 93 KB)
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Regulatory T cells were purified from frozen cord blood samples using an anti-CD25 isolation procedure previously described. (A) Representative example of flow cytometric phenotyping of cord blood Treg before and after purification. Cord blood contains an easily identifiable CD4+CD25bright population that can be isolated in high purity. The purified cells are largely CD127− and Foxp3+. (B-D) Summary of frozen cord blood preparations (n=12; initial: open bars and purified: closed bars) showing % CD4+, % CD25+Foxp3+ (CD4-gated), and % CD127-Foxp3+ (CD4-gated) respectively.
- Figure S2. Treg expanded with cell-based aAPC have increased survival and accumulation in in vitro suppression assays (JPG, 46.4 KB)
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Cord blood Treg were expanded in vitro using bead-or cell-based aAPC with or without 4.1BBL or OX40L coŽstimulation, then CFSE-labeled and incubated with allogeneic PBMC and anti-CD3 beads for 4 days and Treg cell division. Quantitation of Responder Frequency (A) and Proliferative Capacity (# of divisions for proliferating cells) (B) at Treg:PBMNC ratios of 1:2 and 1:16. Data are quantitation of three independent experiments.
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