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Blood, Vol. 112, Issue 13, 4971-4980, December 15, 2008
Stimulation of dendritic cells via the dectin-1/Syk pathway allows priming of cytotoxic T-cell responses
Blood LeibundGut-Landmann et al.
112: 4971
Supplemental materials for: LeibundGut-Landmann et al
Files in this Data Supplement:
- Figure S1. Plasmacytoid DC respond poorly to curdlan (JPG, 68.8 KB)
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(A) Bone marrow cells that were grown in vitro in the presence of Flt3L were stimulated with curdlan (50 µg/ml) or CpG (0.5 µg/ml) for 18 hours. Brefeldin A was added for the last 5 hours. IL-12 p40 staining of the pDC (CD11c+CD11b−B220+) and conventional DC populations (CD11c+CD11b−B220+) is shown. Numbers represent percentage of events in the indicated gate. (B) Expression of Dectin-1 on pDC and conventional DC generated in vitro in the presence of Flt3L. Cells were stained with anti–Dectin-1 (thick line) or rat IgG2b (solid grey). DC subtypes were defined by gating on CD11c+CD11b−B220+ (pDC) and CD11c+CD11b−B220+ cells (conventional DC) respectively. Numbers indicate mean fluorescence intensity (GeoMFI).
- Figure S2. The adjuvant effect of curdlan is MyD88-independent (JPG, 53.2 KB)
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MyD88-deficient and C57Bl/6 control mice were immunized in both hind footpads with OVA protein alone (none) or together with curdlan as indicated. Target cells were injected six days later and mice were analyzed on day 7. (A) The frequency of SIINFEKL-H2Kb tetramer+ CD8+ splenocytes is shown. Each dot represents an individual mouse from two pooled experiments. (B) In vivo CTL activity measured by target cell elimination. Individual mice from 2 pooled experiments are shown. Differences between C57BL/6 and myd88−∕− groups in both (A) and (B) were not statistically different (p>0.05).
- Figure S3. IL-12 p70 is not essential for CTL priming in vivo (JPG, 52.3 KB)
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IL-12 p35-deficient and C57Bl/6 control mice were immunized in both hind footpads with OVA protein alone (none) or together with curdlan as indicated. Target cells were injected six days later and mice were analyzed on day 7. (A) The frequency of SIINFEKL-H2Kb tetramer+ CD8+ splenocytes is shown. Each dot represents an individual mouse from two pooled experiments. (B) In vivo CTL activity measured by target cell elimination. Individual mice from 2 pooled experiments are shown. Differences between C57BL/6 and p35−∕− groups in both (A) and (B) were not statistically different (p>0.05).
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