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Blood, Vol. 109, Issue 8, 3333-3341, April 15, 2007
Essential role of the TNF-TNFR2 cognate interaction in mouse dendritic cell–natural killer cell crosstalk
Blood Xu et al.
109: 3333
Supplemental materials for: Xu et al, Vol 109, Issue 8, 3333-3341
Files in this Data Supplement:
- Figure S1. fmDC TNF induces IFN-γ secretion in fNK cells by engaging NK cell TNFR2 (PDF, 25.6 KB) -
Wild-type (wt) or TNF–/– (A), TNFR1–/– (B), and TNFR2–/– (C) fiDCs and fNK cells were cultured for 24 hours in the presence of LPS either alone or in the following combinations: wild-type+wild-type, gene-deficient+gene-deficient, wild-type+gene-deficient, and gene-deficient+wild-type. Following this incubation, cell culture–conditioned media were assessed by IFN- ELISA. The data are from a representative experiment of 2 to 5 similar experiments performed. They represent means ± SD of triplicates IFN- ng/mL/0.5 × 106 cells. The asterisks indicate the statistical significance of data as in Figure 6B-G.
- Figure S2. ciDC–caNK cell crosstalk via TNF mediates increases in expression of CD40 on DCs (PDF, 38.1 KB) -
Wild-type (wt) or TNF–/– (T–/–) ciDCs and caNK cells were cultured for 24 hours in the presence of 6000 IU/mL IL-2 either alone or in the following combinations and 3:1 ratio: wild-type+wild-type and TNF–/–+TNF–/– and compared for the expression of CD40 using flow cytometry. The data are log10 fluorescence obtained with isotype-matched nonreactive control antibodies (filled histograms), DCs cultured alone stained with anti-CD40 mAb (thin-line histograms), and DCs+NK cells cocultured stained with anti-CD40 mAb (thick-line histograms). The numbers in parentheses are MFI.
- Figure S3. DC–NK cell crosstalk via TNF mediates increases in DC secretion of IL-12p70 (PDF, 12.2 KB) -
Wild-type (wt) or TNF–/– bone marrow–derived ciDCs propagated for 9 days in the presence of GM-CSF/IL-4 and caNK cells expanded in vitro for 6 days were cultured for 48 hours in the presence of 2 µg/mL LPS and 6000 IU/mL IL-2 either alone or cocultured in the following combinations and 1:1 ratio: wild-type+wild-type and TNF–/–+TNF–/– and their cell culture–conditioned media examined for the presence of IL-12p70 using ELISA. The data are from a representative experiment of 2 similar experiments performed. They are means ± SD of triplicates IL-12p70 ng/mL/0.5 × 106 cells. Single asterisk indicates significant increases of IL-12 in wtDC+wtNK cell coculture in comparison with wtDC or wtNK cell cultures alone. Double asterisks indicate significant decreases of IL-12 in TNF–/–DC+TNF–/–NK cell cocultures in comparison with wtDC+wtNK cell cocultures.
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