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Blood, Vol. 95 No. 11 (June 1), 2000:
pp. 3396-3402
Binding and transfer of verocytotoxin by polymorphonuclear
leukocytes in hemolytic uremic syndrome
D. Maroeska W. M. te Loo,
Leo A. H. Monnens,
Thea J. A. M. van der Velden,
Mario A. Vermeer,
Frank Preyers,
Pierre N. M. Demacker,
Lambertus
P. W. J. van den Heuvel, and
Victor W. M. van Hinsbergh
From the Department of Pediatrics, University Hospital Nijmegen,
The Netherlands; Gaubius Laboratory TNO-PG, Leiden, The
Netherlands; Institute for Cardiovascular Research, Vrije Universiteit,
Amsterdam, The Netherlands; Department of Hematology, University
Hospital Nijmegen, Nijmegen, The Netherlands; and the
Department of General Internal Medicine, University Hospital Nijmegen,
Nijmegen, The Netherlands.
The hemolytic uremic syndrome (HUS) is the most common cause of
acute renal failure in children. The role of a verocytotoxin (VT)-producing Escherichia coli has been strongly implicated in the epidemic form of HUS. Although direct toxicity of VT on glomerular endothelial cells has been demonstrated, it remained still unclear how
the VT is transported from the intestine to the target organs. In this
study we demonstrate that VT, when incubated in whole blood, binds
rapidly and completely to human polymorphonuclear leukocytes (PMNs) and
not to other components of blood. Binding studies with
125I-VT-1 showed a single class of binding sites on freshly
isolated, nonstimulated human PMNs. The
Kd of VT-binding to PMNs was
10-8 mol/L, 100-fold less than that of the VT-receptor
globotriaosylceramide. On incubation of VT-preloaded PMNs with human
glomerular microvascular endothelial cells (GMVECs), transfer of VT-1
to the endothelial cells occurred. Incubation of nonstimulated GMVECs
with VT-preloaded PMNs, but not with PMNs or VT-1 alone, caused
inhibition of protein synthesis and cell death. Our data are in concert
with a role of PMNs in the transfer of VT from the intestine to the
kidney endothelium. This transfer occurs by selective
binding to a specific receptor on PMNs and subsequent passing of the
ligand VT to the VT-receptor on GMVECs, which causes cell damage. This
new mechanism further underpins the important role of PMNs in HUS.
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