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Verotoxin-1 promotes leukocyte adhesion to cultured endothelial cells under
physiologic flow conditions
M Morigi, G Micheletti, M Figliuzzi, B Imberti, MA Karmali, A Remuzzi, G Remuzzi and C Zoja
Mario Negri Institute for Pharmacological Research, Bergamo, Italy.
Hemolytic uremic syndrome (HUS), which is the most common cause of acute
renal failure in infants and small children, is caused by verotoxin
(VT)-producing Escherichia coli infection. Endothelial injury determines
microvascular thrombosis and evidence is available from recent studies that
suggests that leukocyte activation participates in endothelial damage. We
studied here the effect of VT-1 on leukocyte adhesion to vascular
endothelium under physiologic flow conditions. Human umbilical vein
endothelial cells (HUVECs) were incubated for 24 hours with VT-1 (0.1, 1,
and 10 pmol/L) and then exposed to a total leukocyte suspension in a
parallel plate flow chamber under laminar flow conditions (1.5 dynes/cm2).
Adherent cells were counted by digital image processing. Results showed
that VT-1 dose-dependently increased the number of adhering leukocytes to
HUVECs as compared with unstimulated cells. The adhesive response elicited
by VT-1 was comparable to that of interleukin-1 beta (IL-1 beta), one of
the most potent inducers of endothelial cell adhesiveness. Exposure of
HUVECs to VT-1 did not affect the number of rolling leukocytes, which was
similar to that of control values. To examine the role of adhesion
molecules in VT-1-induced leukocyte adhesion, HUVECs were incubated with
mouse monoclonal antibodies against E-selectin, intercellular adhesion
molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1) before
adhesion assay. Functional blocking of E-selectin, ICAM-1, and VCAM-1 on
endothelial cells significantly inhibited VT-1-induced increase in
leukocyte adhesion. In some experiments, before VT-1 incubation, HUVECs
were pretreated for 24 hours with tumor necrosis factor alpha (TNF alpha;
100 U/mL), which is known to increase VT receptor expression on HUVECs. The
number of adhering leukocytes on HUVECs exposed to TNF alpha and VT-1
significantly increased as compared with HUVECs incubated with VT-1 and TNF
alpha alone. These results suggest that VT-1 modulates
leukocyte-endothelium interaction, thus increasing leukocyte adhesion and
upregulating adhesive proteins on endothelial surface membrane.
Volume 86,
Issue 12,
pp. 4553-4558,
12/15/1995
Copyright © 1995 by The American Society of Hematology
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