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Blood, Vol. 113, Issue 23, 5811-5818, June 4, 2009
PD-1 on dendritic cells impedes innate immunity against bacterial infection
Blood Yao et al.
113: 5811
Supplemental materials for: Yao et al
Files in this Data Supplement:
- Figure S1. Rag-1KO and Rag-1 PD-1 DKO mice have no significant difference in their cellular components (JPG, 45.4 KB)
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Cells from spleen, liver, and total PBMC were acquired from 7–8 weeks Rag-1KO (n=5) and DKO (n=4) mice. NK cells, DCs, and neutrophils subpopulation were measured by FACS analysis.
- Figure S2. B7-H1 blockade enhances innate resistance to LM infection (JPG, 32.3 KB)
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(A) B6 Rag-1KO mice were i.p. injected with 250ug control hamster Ig or anti-mouse B7-H1(10B5) one day before and one day after 105 CFU LM i.v. infection. Survival was monitored for 14 days. P<0.05 between the two groups. (B) Same procedure as in (A), 3 days post LM infection, liver LM counts (CFU per gram tissue) were determined. * P<0.05.
- Figure S3. No PD-1 expression was detected on plasmacytoid DCs (JPG, 52.7 KB)
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DCs were purified from Rag-1KO or DKO spleens using CD11c MACS beads, and incubated with 5 × 104 CFU/ml LM in vitro for two days. pDCs were further gated by I-Abmedium CD11cmedium PDCA-1+ markers, and stained for PD-1 expression.
- Figure S4. B7-H1 was upregulated in mouse splenocytes post LM infection (JPG, 71.3 KB)
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Naïve and 24hrs LM infected B6 Rag-1KO spleens were harvested. B7-H1 expression was upregulated in all DX5+, Gr-1+, CD11b+, and CD11c+ subpopulations of splenocytes.
- Figure S5. PD-1 expression is modulated by different TLR ligands and cytokines (JPG, 59.8 KB)
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Splenic CD11c+ cells were purified using CD11c MACS beads from Rag-1KO and DKO mice, incubated in vitro for 48hrs with different TLR ligands and cytokines. Percentage of PD-1 positive cells was shown after stimulation.
- Figure S6. Naïve Rag-1KO DCs and DKO DCs have similar intrinsic ability to produce IL-12 and TNFα upon HKLM stimulation (JPG, 75.3 KB)
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Naïve B6 Rag-1KO and Rag-1 PD-1 DKO spleen cells were stimulated with PBS or HKLM in the presence of brefeldin A for 4 hrs. I-Ab+ CD11c+ gated DCs were stained for intracellular IL-12p70 (A) and TNF-α (B).
- Figure S7. Adoptive transfer of PD-1 deficient DCs increased host serum IL-12 level post LM challenge (JPG, 21.7 KB)
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Rag-1KO hosts received PBS control or 6 × 106 purified splenic DCs from Rag-1KO or DKO mice, then i.v. challenged with 105 CFU LM. Sera were collected 24 hours post infection. Serum IL-12p70 was determined by sandwich ELISA. * P<0.05.
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