Interstitial movement of T cells within lymph nodes
| Recipient mice . | Genotype of donor T cells . | |||||
|---|---|---|---|---|---|---|
| WT . | Rac1T . | WT . | Rac2−/− . | WT . | DKO . | |
| C57BL/6 | ||||||
| Mean velocity, μm/min | 15.17 ± 0.28 | 14.47 ± 0.24 | 14.63 ± 0.30 | 12.00 ± 0.31* | 14.49 ± 0.31 | 5.6 ± 0.18* |
| Median instantaneous velocity, μm/min | 12.83 ± 0.15 | 12.81 ± 0.12 | 11.00 ± 0.14 | 9.77 ± 0.11* | 12.88 ± 0.18 | 4.2 ± 0.05* |
| Turning angle, degree | 43.81 ± 2.15 | 44.68 ± 2.61 | 46.7 ± 2.72 | 46.17 ± 2.99 | 42.95 ± 3.11 | 97.05 ± 3.43* |
| Motility coefficient, μm2/min | 65.86 | 84.26 | 60.74 | 44.18 | 64.86 | 1.29 |
| plt/plt | ||||||
| Mean velocity, μm/min | 11.57 ± 0.47 | 11.97 ± 0.31 | 10.35 ± 0.21 | 10.49 ± 0.25 | nd | nd |
| Recipient mice . | Genotype of donor T cells . | |||||
|---|---|---|---|---|---|---|
| WT . | Rac1T . | WT . | Rac2−/− . | WT . | DKO . | |
| C57BL/6 | ||||||
| Mean velocity, μm/min | 15.17 ± 0.28 | 14.47 ± 0.24 | 14.63 ± 0.30 | 12.00 ± 0.31* | 14.49 ± 0.31 | 5.6 ± 0.18* |
| Median instantaneous velocity, μm/min | 12.83 ± 0.15 | 12.81 ± 0.12 | 11.00 ± 0.14 | 9.77 ± 0.11* | 12.88 ± 0.18 | 4.2 ± 0.05* |
| Turning angle, degree | 43.81 ± 2.15 | 44.68 ± 2.61 | 46.7 ± 2.72 | 46.17 ± 2.99 | 42.95 ± 3.11 | 97.05 ± 3.43* |
| Motility coefficient, μm2/min | 65.86 | 84.26 | 60.74 | 44.18 | 64.86 | 1.29 |
| plt/plt | ||||||
| Mean velocity, μm/min | 11.57 ± 0.47 | 11.97 ± 0.31 | 10.35 ± 0.21 | 10.49 ± 0.25 | nd | nd |
Purified WT EYFP+ T cells and EYFP+ T cells from Rac1T, Rac2−/−, or DKO mice were labeled with CFSE or CMTMR, mixed, and transferred into C57BL/6 or plt/plt recipient mice. Interstitial movement of labeled T cells within the popliteal lymph node was analyzed 15-72 hours later by multiphoton intravital video microscopy. All results are presented as a pairwise comparison of WT with Rac1T, Rac2−/−, or DKO T cells. In C57BL/6 recipients WT v Rac1T: 13 videos analyzed from 6 independent experiments; WT v Rac2−/−: 14 videos analyzed from 9 independent experiments; WT v DKO: 7 videos analyzed from 4 independent experiments. In plt/plt recipients WT v Rac1T: 6 videos analyzed from 2 independent experiments; WT v Rac2−/−: 11 videos analyzed from 3 independent experiments.
CFSE indicates carboxyfluorescein succinimidyl ester; DKO, double knockout; EYFP, enhanced yellow fluorescent protein; nd, not determined; WT, wild-type.
P < .001.