Impaired homing of ezrin T567E transgenic T cells to lymph node. (A) In vivo homing to lymph node. To assess homing in an internally controlled manner, 2 different T-cell preparations were stained with different levels of CFDA, mixed, injected intravenously, and their recovery in lymph node and blood was assessed by flow cytometry after 1 hour. Data are mean ± SEM for pairs of mice in an experiment representative of 4 performed in which the experimental mix (right side of panel) was ezrin T567E transgenic cells and WT cells. The control mix (left side of panel) was WT cells and WT cells. Recovery ratios for the control mix were all close to 1.0 (dashed horizontal line). In the experimental mix, the recovery ratio was decreased for transgenic T cells in lymph node (P < .05) and increased in blood (P < .01), but recovery ratios close to 1.0 were observed for the non-T cells. (B) Assessment of lymphocyte rolling in inguinal lymph node. Lymph node T cells were injected intra-arterially, and the number showing slow movement characteristic of rolling during the first pass through the lymph node was scored blind. The venule order designations were assigned by counting successive generations of venular branches in the upstream direction from the large collecting venule that drains into the superficial epigastric vein (with first being the largest and fifth being the smallest). (C) Same as in panel B but showing assessment of the fraction of rolling cells that bind for more than 30 seconds.