Intravascular Immune Surveillance by CXCR6+ NKT Cells Patrolling Liver Sinusoids (original) (raw)
Figure 2
CXCR6+ Lymphocytes Patrol Liver Sinusoids
(A) Select confocal microscopic images from intravital videos of liver of an anesthetized cxcr6gfp/+ mouse (40× magnification). CD1d-reactive cells (bright green) can be seen migrating along hepatic sinusoids at an average speed of 16 μm/min. Scale bar is 25 μm. Cell tracks were traced and quantified using Volocity cell-imaging software. Note tracks of cells traveling in opposite directions in the same sinusoid (left side of image, at 0–5 versus 7–12 min).
(B) Velocity quantification of GFP+ lymphocytes. Videos of liver in cxcr6gfp/+ and cxcr6gfp/gfp mice (10× magnification) were analyzed for the velocity of GFP+ cells in 640 cell migration tracks for cxcr6gfp/+ mice and 574 cell migration tracks for cxcr6gfp/gfp mice. Results demonstrate similar velocities of cells with the cxcr6gfp/+ (filled bars, average velocity 16.5 ± 8.3 μm/min) and cxcr6gfp/gfp genotypes (unfilled bars, average velocity 18.4 ± 9.5 μm/min).
(C) Analysis of directedness of cell migration. The same cell tracks as in (B) were analyzed for ratio of overall cell displacement to stepwise-summed path length. Results demonstrate similar degrees of directed crawling by cells from cxcr6gfp/+ (filled bars, average 0.44 ± 0.31) and cxcr6gfp/gfp (unfilled bars, average 0.42 ± 0.30) mice.
(D) Analysis of crawling relative to blood flow in peri-central vein areas of cxcr6gfp/+ and cxcr6gfp/gfp mice. Histograms represent the frequency of cell movements made towards or away from nearby draining areas in likely close proximity to central veins (solid blue curve). The radial step of a cell is the cellular displacement along the axis defined by the cell's initial starting point to the central vein. The same step distances were assigned random orientations and the resultant data were plotted (red dashed curves).