Direct observations of the kinetics of migrating T-cells suggest active retention by endothelial cells with continual bidirectional migration



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).



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