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Blood, Vol. 112, Issue 13, 5037-5045, December 15, 2008
Dendritic cell microvilli: a novel membrane structure associated with the multifocal synapse and T-cell clustering
Blood Fisher et al.
112: 5037
Supplemental materials for: Fisher et al
Files in this Data Supplement:
- Figure S1. Quantifying spatial relationships of DCs and T cells (JPG, 30.2 KB)
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The spatial relationship of any cell pair was quantified based on the mean T-cell radius (rT) and mean DC radius (rD). For a T cell bound to a DC, A ≤ rT+rD; when two T cells interact, B ≤ rT1+rT2; when two separated T cells bind a DC, C > rT1+rT2; for a T cell separate from the DC, D > rT+rD. The mean radius of each cell was calculated by Metamorph software as the average distance from the center of cell to all pixels at its edge.
- Figure S2. CD80 and HLA-DR molecules are enriched at the DC–T-cell interaction site (JPG, 41.2 KB)
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DCs were incubated with allogeneic pan-T cells for six hours when CD80 was stained red, HLA-DR blue, and cytoplasm green. Areas expressing both CD80 and HLA-DR appear purple. (A) A DC (larger cell, left) interacting with a single T cell (right); (B) cytoplasm digitally removed from A for better visualization of HLA–DR and CD80 molecules. Depth of confocal images = 1 µm. Bar = 5 µm.
- Video 1. DC veils are highly motile (MOV, 1.14 MB)
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Differential interference contrast micrograph video of human dendritic cells at 37°C recorded at one frame per second and displayed at fifteen frames per second; motion of veils is clearly visible.
- Video 2. Interaction of T cells with a DC veil (MOV, 4.87 MB)
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Human DCs were stained with cytoplasmic dye CMFDA (green) and incubated with allogeneic T cells stained with DiD (red) and recorded at 37°C by confocal microscopy. One frame was recorded every five seconds and displayed at fifteen frames per second.
- Video 3. Preferential binding of T cells to a single site on the DC (MOV, 3.17 MB)
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Human DCs were incubated with allogeneic CD4+ T cells (green) or CD8+ T cells (red) at 37°C and analyzed by light transmission and fluorescence microscopy combined. Recorded at one frame per minute and displayed at fifteen frames per second.
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