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Blood, Vol. 106, Issue 8, 2818-2826, October 15, 2005
Monocyte-derived dendritic cells activated by bacteria or by bacteria-stimulated epithelial cells are functionally different
Blood Rimoldi et al.
106: 2818
Supplemental materials for: Rimoldi et al, Vol 106, Issue 8, 2818-2826
Files in this Data Supplement:
- Figure S1. Monocyte-derived dendritic cells (MoDCs) downregulate the expression of CD14 and upregulate the expression of CD11c (PDF, 22.1 KB) -
MoDCs were incubated for 5 days with DC-differentiation cytokines GM-CSF and IL-4. Quality of MoDC preparation was assessed by cytofluorometry after staining with CD14, CD1a, and CD11c antibodies. Open histograms represent isotype controls; filled histograms, antibody staining.
- Figure S2. The integrity of the epithelial barrier is preserved in the 5-hour experimental procedure (PDF, 24.9 KB) -
Monolayers of Caco-2 cells were infected apically (time 0) with SL-WT in medium without antibiotics. One hour after infection, bacteria were washed out and medium was changed with one containing gentamicine (100 µg/mL). Transepithelial resistance was measured every hour from bacterial infection. Data are shown as means (± SD) of six different transwells.
- Figure S3. Indirectly activated MoDCs release in IL-40 but not IL-12p70, even after 16 hours’ exposure of ECs to bacteria (PDF, 72.0 KB) -
(Left) MoDCs were activated with S typhimurium (SL-WT noninvasive SL-InvA, nonflagellated SL-FIIC) or L plantarum (LP). (Right) MoDCs were incubated with supernatants of ECs incubated or not for 16 hours with bacteria (4 × 106 CFUs/transwell) from the apical face. Cytokine release was measured in culture supernatants by ELISA. The difference of IL-12 production between bacteria and EC/bacteria-activated MoDCs is highly significant (*P < .01). Error bars indicate means ± SD.
- Figure S4. EC-conditioned MoDCs release IL-10 but not IL-12p70, even after exposure to EC monolayers and apical bacteria (PDF, 85.2 KB) -
MoDCs were treated for 24 hours as follows: (left) direct system, MoDCs were seeded facing the basolateral membrane of EC monolayer. Bacteria were incubated from the apical face. (Right) Conditioning plus direct system, MoDCs were incubated with supernatants of ECs nonincubated with bacteria for 16 hours, subsequently conditioned MoDCs were seeded facing the basolateral membrane of EC monolayer. Bacteria were incubated from the apical face. Cytokine release was measured in culture supernatants by ELISA. The difference of IL-12 production between directly and indirectly activated MoDCs is significant (*P < .05). Error bars indicate means ± SD.
- Table S1. Total number of bacterial CFUs present in the lower face medium of transwell coculture systems at 16 hours from bacterial apical treatment (PDF, 47.0 KB)
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