Arrest efficiency of Kindlin-3–deficient effectors on inflamed lymph nodes vessels is lower compared with wt effectors (A) Accumulation efficiency of coinjected CFSE wt and CMTMR-labeled Kindlin-3 mutant effector T cells inside resting and inflamed inguinal lymph node vessels as determined by multiphoton intravital microscopy at 40 minutes after intracardiac injection. CFA was injected subcutaneously 48 hours before adoptive transfer of T cells. Blood vessels were visualized by Qtracker655 quantum dots. Each dot represents a single field of view (0.23 mm²). For each experimental group, the number of accumulated T cells per field was normalized to the number of T cells in the corresponding group found to circulate in the blood (ie, the flux of freely flowing T cells). Each colored dot represents a single field of view. Results are accumulated from 3 independent experiments. ***P < .0001. (B) Effect of pretreatment of wt effector T cells with either LFA-1–blocking mAb (left), α₄-blocking mAb (middle), with a nonblocking anti-CD45 mAb (all panels), or a nonbinding mAb (control, right panel) on accumulation of wt effector T cells inside inflamed lymph nodes 40 minutes after intracardiac injection. Differently dye-labeled T cells were pretreated with either mAb 20 minutes before injection, washed, mixed, and coinjected at a 1:1 ratio. Each colored dot represents a single field of view. The numbers of cells were normalized to the T-cell flux as in panel A. Results are accumulated from 3 independent experiments. ***P < .0001. (C) Effect of PTx on the accumulation of wt effector T cells inside inflamed lymph nodes 40 minutes after intracardiac injection. Sham-treated CFSE-labeled T cells were coinjected with PTx-pretreated CMTMR-labeled T cells, as in Figure 3C. Each dot represents a single field of view. The numbers of cells were normalized to the T-cell flux as in panels A-B. One experiment is representative of 3 experiments. (D) Effect of LFA-1 blocking on early accumulation (40 minutes after intracardiac injection) of Kindlin-3 null T cells in inflamed lymph node vessels. Differently labeled Kindlin-3–deficient effector T cells were pretreated with either LFA-1–blocking mAb (left), α₄-blocking mAb (right), or with a nonblocking anti-CD45 mAb, as in panel B. Each dot represents a single field of view. The numbers of cells were normalized to the T-cell flux as in panel A. **P < .0032; ***P < .0001; n.s., not significant.