von Andrian Lab Videos
 

 

BONE MARROW IS A MAJOR RESERVOIR AND SITE OF RECRUITMENT FOR CENTRAL MEMORY CD8+ T CELLS

Immunity Supplementary Movie s1.mp4

Three-dimensional rotation (360 degrees) of a digitally rendered z-stack of MP-IVM images showing transmigrated TCM (blue) in a skull BM cavity (red). BM vessels arecontrasted with 2 MD FITC-dextran (green).  Surrounding bone is black.  For further information refer to fig. 2F.

 


Immunity Supplementary Movie s2.mp4

Three-dimensional time-lapse video demonstrating TCM (blue) migration within the extravascular space in BM cavities delineated by rhodamine 6G (red).  The intravascular compartment was contrasted by 2 MD FITC-dextran (green).  Image stacks (23 optical slices, 5 µmspacing) were acquired at one stack/min.  Video playback is at 15 frames per second (i.e. time-lapse is 900x).


Immunity Supplementary Movie s3.mp4

CFCE-labeled TCM in BM microvessels of an anesthetized mouse.  Cells were injected into the feeding carotid artery, and their passage through BM microvessels in the skull was recorded by video-triggered stroboscopic illumination.  This movie is shown in real-time (30 frames per s).  Cells are seen to roll in venules and sinusoids.  Some of the rolling cells adhere firmly and may eventually emigrate into the surrounding BM cavities.


Immunity Supplementary Movie s4.mp4

A three-dimensional rotation (30 degrees) showing the distribution of control (blue) and PTX-treated (red) TCM in the intra- and extravascular compartment in a BM cavity.  BM vessels are filled with 2 MD FITC-dextran (green).  Note that while the majority of control cells extravasated (transmigration efficiency in this field of view was3.7), PTX-treated cells remained mostly confined to the intravascular compartment (transmigration efficiency 1.8).


For details on experimental methods & techniques see:
Mazo, I.B., Honczarenko, M., Leung, H., Cavanagh, L.L., Bonasio, R., Weninger, W., McEver, R., Koni, P.A., Silberstein, L.E. and von Andrian, U.H., Bone marrow is a major reservoir and site of recruitment for central memory CD8+ T cells. Immunity. 22:259-270, 2005 [Full Text PDF] [Video Legend]