Figure 4.
Figure 4. T-cell contact induces a stop signal in B cells. B-T pairs are pushed by T-cell movements and maintain a fixed contact plane. Naive OVA-loaded B cells were stained with CFSE and embedded within 3-D collagen together with fresh DO11.10 T cells. (A) A B cell is observed establishing 2 successive contacts to 2 distinct T cells and is only motile in the intervening time, whereas no T-cell contact is maintained (Supplemental Movie 6). An established B-T pair moves through the collagen pushed forward by T-cell movements (red path). Twenty-two minutes later, the B-T pair breaks up, and the B cell now moves by itself for the next 160 minutes (white path). Then the B cell is caught by a moving B-T pair (red arrowhead), whereby the T cell exchanges the previously contacted B cell for the new one. Again, the previously motile B cell stops all autonomous migration and is transported passively by the motile T cell (red path) 30 minutes after culture set-up. (B) Cell tracking analysis of the velocities of the cell-cell pairs and the free B cell during the events described in panel A. The B cell develops autonomous migration immediately after breaking of the contact to a T cell (arrow) and ceases migration immediately after engagement by another T cell (arrow). B-T pairs move at velocities of free T cells.7,10 (C) An exceptionally motile B-T pair covering a large distance (white path) within 116 minutes of observation. Note that the path makes many twists and turns (Supplemental Movie 7). Day 2 after culture set-up. (D) A close inspection of a motile B-T pair shows that, despite the pair's many turns (white path), the B cell (green) remains round and forms a fixed unit with the leading edge of the T cell. The uropod of the T cell is often at an angle relative to the B-T contact plane (yellow line). These movements of the T-cell uropod appear to generate the force for the movement of the entire B-T pair (Supplemental Movie 8). The lower row of panel D is an enhanced view of the cell-cell pair in the red box in the upper row of the panel. Day 3 after culture set-up. Scalebar: 50 μm (A,D), 100 μm (B). Numbers indicate minutes of real time.

T-cell contact induces a stop signal in B cells. B-T pairs are pushed by T-cell movements and maintain a fixed contact plane. Naive OVA-loaded B cells were stained with CFSE and embedded within 3-D collagen together with fresh DO11.10 T cells. (A) A B cell is observed establishing 2 successive contacts to 2 distinct T cells and is only motile in the intervening time, whereas no T-cell contact is maintained (Supplemental Movie 6). An established B-T pair moves through the collagen pushed forward by T-cell movements (red path). Twenty-two minutes later, the B-T pair breaks up, and the B cell now moves by itself for the next 160 minutes (white path). Then the B cell is caught by a moving B-T pair (red arrowhead), whereby the T cell exchanges the previously contacted B cell for the new one. Again, the previously motile B cell stops all autonomous migration and is transported passively by the motile T cell (red path) 30 minutes after culture set-up. (B) Cell tracking analysis of the velocities of the cell-cell pairs and the free B cell during the events described in panel A. The B cell develops autonomous migration immediately after breaking of the contact to a T cell (arrow) and ceases migration immediately after engagement by another T cell (arrow). B-T pairs move at velocities of free T cells.7,10  (C) An exceptionally motile B-T pair covering a large distance (white path) within 116 minutes of observation. Note that the path makes many twists and turns (Supplemental Movie 7). Day 2 after culture set-up. (D) A close inspection of a motile B-T pair shows that, despite the pair's many turns (white path), the B cell (green) remains round and forms a fixed unit with the leading edge of the T cell. The uropod of the T cell is often at an angle relative to the B-T contact plane (yellow line). These movements of the T-cell uropod appear to generate the force for the movement of the entire B-T pair (Supplemental Movie 8). The lower row of panel D is an enhanced view of the cell-cell pair in the red box in the upper row of the panel. Day 3 after culture set-up. Scalebar: 50 μm (A,D), 100 μm (B). Numbers indicate minutes of real time.

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