Data Supplement

Files in this Data Supplement:

• Video 1a (MOV, 718 Kb)
  - Time-lapse videos (accelerated 150x) showing an example of mature, bone marrow derived, wt (a) and WASp- (b) DCs in the presence of OT-1 cells. DCs were pulsed with 0,1 nM of OVA peptide before recording. In the wt sequence the DC actively capture the T cell thanks to its membrane extensions and cell body movements. In the WASp sequence the DC, despite its long membrane projections, fails to move toward the T cells and even after several short contacts mediated by dendrites a stable interaction is not formed.
• Video 1b (MOV, 904 Kb)
  
• Video 2a (MOV, 1.25 Mb)
  - Time-lapse videos videos (accelerated 150x) showing examples of endogenous DCs isolated from LNs of wt (a) or WASp- (b) mice. Cells were pulsed with peptide and co-cultured in the presence of OT-I. The arrow in movie 3a indicates a DC that travel toward a T cell to capture it and form a stable contact. Movie 3b illustrates an example of failure to stabilize the contact by a WASp-DC.
• Video 2b (MOV, 818 Kb)
  
• Video 3a (MOV, 883 Kb)
  - OVA specific OT-I cells labeled with CMTMR were injected i.v. into wt or WASp- mice that were immunized 4 h later with 0,1 ug of anti-DEC205-OVA conjugates plus anti CD40. Two-photon imaging was performed 18 h later on draining LNs. a) One representative example of trajectories of T cells in LN of wt mice. b) As above, T cells in LNs of WASp animals. Time of imaging: 30 min. Playback speed: 200. Size 175x175 um.
• Video 3b (MOV, 1.92 Mb)
  
• Video 4a (MOV, 3.69 Mb)
  - OVA specific OT-I cells labeled with CFSE were adoptively transferred into wt recipient mice that received subcutaneous injection of CMTMR labeled peptide pulsed wt (a) or WASp- (b) DCs in the footpad. Popliteal LNs were collected 20h post injection and DC-T interactions were imaged during a 30 minutes session.
• Video 4b (MOV, 2.64 Mb)