Table 1: Characteristics of migration of lymphocytes migrating through endothelial cell monolayers on clear substrates
|
Treatment/Cell |
Cells undergoing |
Cells undergoing |
Transit interval |
Migration velocity |
|
1 or more transits (%) |
more than 1 transit (%) |
(min) |
(μm∕min) | |
|
A. Migration of PBL |
through EC exposed to different cytokines - Static assay (n=4) | |||
|
TNF |
18.3 ± 8.6 |
6.6 ± 3.8 |
4.4 ± 0.9 |
6.4 ± 0.2 |
|
IFN |
25.9 ± 5.6 |
7.5 ± 3.9 |
4.4 ± 0.5 |
10.5 ± 1.4 |
|
TNF+IFN |
21.5 ± 4.3 |
9.6 ± 4.2 |
3.9 ± 0.2 |
8.9 ± 1.0 |
|
B. Different lymphocyte subsets migrating through |
EC treated with TNF+INF - |
Static assay (n=3) | ||
|
PBL |
17.5 ± 5.6 |
7.1 ± 3.4 |
3.6 ± 0.5 |
8.8 ± 2.4 |
|
CD3+ |
15.9 ± 6.0 |
5.2 ± 3.4 |
4.6 ± 0.8 |
6.1 ± 0.3 |
|
CD4+ |
19.2 ± 2.8 |
5.6 ± 2.9 |
3.8 ± 0.8 |
7.1 ± 1.6 |
|
C. Migration of PBL through EC exposed to different cytokines - Flow-based assay (n=4) | ||||
|
TNF |
23.9 ± 1.8 |
11.1 ± 3.8 |
3.7 ± 0.5 |
5.8 ± 0.5* |
|
IFN |
31.5 ± 4.0 |
13.7 ± 4.1 |
3.9 ± 0.5 |
9.6 ± 1.5 |
|
TNF+IFN |
32.5 ± 5.2 |
13.6 ± 7.9 |
4.4 ± 1.0 | |
|
D. Migration of PBL |
through EC on collagen gels - |
Static assay (n=3) | ||
|
TNF+IFN |
21.0 ± 2.5 |
7.8 ± 4.1 |
4.2 ± 0.4 |
5.4 ± 0.8 |
Individual lymphocytes were tracked over the 6min period and their location above or beneath the endothelium analysed every 10s. The
percentages of lymphocytes migrating through the monolayer once (1 transit) or migrating back and forth (>1 transit) were determined. The
average interval between transits for those cells that did move between compartments was also determined. The velocity of migration under the
monolayer was measured over a 5-minute period. Data are the mean ± SEM from n independent experiments. * ANOVA showed significant
effect of treatment on migration velocity, with TNF significantly different from IFN by Tukey test (both p<0.05).