Data Supplement

Files in this Data Supplement:

• Supplemental Video 1 (MOV 2.92 Mb)
  - Adherence/detachment behavior of CD3-activated T-cells under physiological shear stress. CD3-activated CD4+ PBL were CSFE-labeled (green), injected into the flow chamber containing a monolayer of HeLa cells (pseudo-colored red) and incubated for 15 min prior to the start of flow. Time-lapse microscopy was used to record differential interference contrast (DIC) and fluorescent images of adherent T-cells as the flow rate of the medium was increased. DIC and fluorescent images were captured using an Olympus IX70 inverted microscope every 15 sec for 17 min at an original magnification of x10. Images were saved as .tif files and pseudo-colored using MetaMorph (Molecular Devices, Downingtown, PA). Movies were made using ImageJ (Freeware, NIH).
• Supplemental Video 2 (MOV 4.36 Mb)
  - Adherence/detachment of CD3-activated T-cells under physiological shear stress in the presence of an anti-β7 antibody. Experimental set up and video generation as under "Supplemental Video 1" but with the addition of a blocking antibody to β7 (10 μg/ml) during the incubation phase.
• Supplemental Video 3 (MOV 943 Kb)
  - Adherence/detachment of CD3/CD46-activated T-cells under physiological shear stress. Experimental set up and video generation as under "Supplemental Video 1" but with the usage of CD3/CD46-activated CD4+ PBL.
• Supplemental Video 4 (MOV 623 Kb)
  - Adherence/detachment of CD3/CD46-activated T-cells under physiological shear stress in the presence of an anti-β7 antibody. Experimental set up as under "Supplemental Video 1" but with the usage of CD3/CD46-activated CD4+ PBL and the addition of a blocking antibody to β7 (10 μg/ml) during the incubation phase.
• Supplemental Video 5 (MOV 320 Kb)
  - CD3/CD46-activated T-cells form homotypic clusters on HeLa cells. Time-lapse microscopy of CD3/CD46-activated T-cells (green) during initial adherence to a monolayer of HeLa cells (pseudo-colored red). CD3/CD46-activated CD4+ PBL were CSFE-labeled, injected (1 x 106) into the flow chamber containing a confluent monolayer of HeLa cells and incubated for 15 min prior to the start of flow. DIC and fluorescent images were captured every 30 sec for 10 min at an original magnification of x10 and movies made as under "Supplemental Video 1".