Companion web page to

A Wülfing, C., Sjaastad, M. D. and Davis, M. M.

Visualizing the dynamics of T cell activation: ICAM-1 migrates rapidly to the T cell: B cell interface and acts to sustain calcium levels

Proc. Natl. Acad. Sci. USA (1998) 95, 6302-7 and

B Wülfing, C. and Davis, M. M.

A receptor/cytoskeletal movement triggered by costimulation during T cell activation

Science (1998) 282, 2266-9

All movies mentioned in the above publications can be accessed from this web page as Quicktime movies with MPEG compression. Any movie player will work. We have good experiences with the program Sparkle Fat that can be obtained free from the Internet, e. g. at http://www.versiontracker.com/. Some of the files are large, so fast internet access helps a lot.

A These movies show the interaction of T cells with different antigen presenting cells or antigen presenting cell substitutes, each presenting a peptide that can be recognized by the T cell receptor of the T cell. The antigen presenting cells are:

Movie 1 B cell lymphoma CH27, transfected with ICAM-1 fused to GFP

Movie 2 CHO cells transfected with MHC II and ICAM-1 fused to GFP

Movie 3 Supported lipid bilayer containing MHC II

The first two movies both consist of two identical bright field panels. Overlaid on the top is a rainbow color representation of the intracellular calcium concentration of the T cell. Blue indicates low, red high calcium concentration. Overlaid on the bottom is a custom color representation of the ICAM-1&endash;GFP fluorescence intensity of the antigen presenting cell. Green indicates low, red high ICAM-1&endash;GFP fluorescence intensity. Movie 3 consist only of one panel that is equal to the top panel of the first two movies.

B These movies show the interaction of T cells with CH27 B cell lymphomas as antigen presenting cells presenting a peptide that can be recognized by the T cell receptor of the T cell. In addition, the movement of the cortical actin cytoskeleton is monitored using large cross-linking beads.

Movie 1 In the absence of pharmacological agents

Movie 2 In the presence of 2 mM butanedione monoxime, a blocker of the myosin II ATPase

The first movie consist of one, the second of two identical bright field panels. Overlaid, in the second movie on the top, is a rainbow color representation of the intracellular calcium concentration of the T cell. Blue indicates low, yellow in the first and red in the second high calcium concentration. Overlaid on the bottom of the second movie is a custom color representation of the ICAM-1&endash;GFP fluorescence intensity of the antigen presenting cell. Green indicates low, red high ICAM-1&endash;GFP fluorescence intensity.