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Department of Immunology, Duke University Medical Center, Durham, NC 27710
Selectin family members largely mediate initial tethering and
rolling of leukocytes on vascular endothelium, whereas integrin and Ig
family members are essential for leukocyte firm adhesion. To quantify
functional synergy between L-selectin and Ig family members during
leukocyte rolling, the EA.hy926 human vascular endothelial line was
transfected with either fucosyltransferase VII (926-FtVII) cDNA to
generate L-selectin ligands alone or together with ICAM-1 cDNA
(926-FtVII/ICAM-1). The ability of transfected 926 cells to support
human leukocyte interactions was assessed in vitro using parallel plate
flow chamber assays. Lymphocyte rolling on 926-FtVII cells was
increased by 
70% when ICAM-1 was expressed at physiological levels.
Although initial tether formation was similar for both cell types,
lymphocyte rolling was 26% slower on 926-FtVII/ICAM-1 cells.
Pretreatment of lymphocytes with an anti-CD18 mAb eliminated the
increase in rolling, and all rolling was blocked by anti-L-selectin
mAb. In addition, rolling velocities of lymphocytes from
CD18-hypomorphic mice were 48% faster on 926-FtVII/ICAM-1 cells, with
a similar reduction in rolling frequency relative to wild-type
lymphocytes. CD18-hypomorphic lymphocytes also showed an
40% decrease in migration to peripheral and mesenteric lymph nodes
during in vivo migration assays compared with wild-type lymphocytes.
Likewise, wild-type lymphocyte migration to peripheral lymph nodes was
reduced by 50% in ICAM-1-/- recipient mice. Similar
to human lymphocytes, human neutrophils showed enhanced rolling
interactions on 926-FtVII/ICAM-1 cells, but also firmly adhered. Thus,
in addition to mediating leukocyte firm adhesion, CD18 integrin/ICAM-1
interactions regulate leukocyte rolling velocities and thereby optimize
L-selectin-mediated leukocyte rolling.
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