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Blood, Vol. 112, Issue 5, 2129-2138, September 1, 2008
In vivo–activated CD103+CD4+ regulatory T cells ameliorate ongoing chronic graft-versus-host disease
Blood Zhao et al.
112: 2129
Supplemental materials for: Zhao et al
Files in this Data Supplement:
- Figure S1. Donor-type CD103+ Treg cells from recipients without chronic GVHD ameliorated ongoing chronic GVHD (JPG, 36.3 KB)
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Sublethally irradiated BALB/c recipients were injected with 100 × 106 DBA/2 donor spleen cells. 20 days after donor cells injection when all the recipients showed severe proteinuria, the recipients were injected with CD103+ Treg cells(1 × 106) from the spleen of recipients given 6.25 × 106 donor spleen cells. The control recipients were injected with PBS only. The recipients were monitored daily for proteinuria and survival. The percentage of recipients with reversal of proteinuria and percentage of survival are shown. There are 8 recipients in each group combined from 2 replicate experiments.
- Figure S2. In vitro activation and expansion of CD25hi natural Treg cells (JPG, 63 KB)
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Sorted CD25hiCD4+ T cells from DBA/2 donors were cultured with anti-CD3/CD28 beads and IL-2 (2000 U/ml) as previously described.1 (A) The percentage of FoxP3+ cells before and after culture. (B) The expansion levels at different days after cell culture.
- Figure S3. Upregulation of granzyme B expression by CD103+ Treg cells from chronic GVHD recipients (JPG, 24.5 KB)
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Spleen cells from donor mice or chronic GVHD recipients 15 days after HCT were stained with anti-CD4, CD25, FoxP3 and granzyme B or anti-CD4, CD103, FoxP3 and granzyme B. The gated CD25hiCD4+FoxP3+ cells from donor (green) and CD103+CD4+FoxP3+ cells from recipient (blue) are shown as a histogram. The isotype control of the two types Treg cells were very similar and were shown in red and black, respectively. One representative of three experiments is shown. The mean fluorescence intensity (mean ± SE) for donor CD25hiFoxP3+ Treg cells and recipient CD103+FoxP3+ Treg cells was 7.8 ± 1.7 versus 22.4 ± 4.2 (P<0.05).
- Figure S4. Induction of donor-type CD103+ Treg cells in chronic GVHD recipients required donor CD25hiCD4+ T cells in graft (JPG, 40.1 KB)
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Whole spleen or CD25+ T depleted spleen cells (100 × 106) were transplanted into sublethally irradiated recipients. 20 days after HCT, recipients spleen cells were stained for anti-CD5.1 (donor-type marker), CD4, CD103, and FoxP3. (A) FoxP3 versus CD103 after gating on CD5.1+CD4+ cells. One representative of 4 experiments is shown. (B) Mean ± SE of percentage of CD103− or CD103+ Treg cells.
REFERENCE
1. Tang Q, Henriksen KJ, Bi M, et al. In vitro-expanded antigen-specific regulatory T cells suppress autoimmune diabetes. J Exp Med. 2004;199:1455–1465.
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