|
|
 Previous Article | Table of Contents | Next Article 
Blood, 15 September 2001, Vol. 98, No. 6, pp. 1828-1835
HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
Verotoxin-1-induced up-regulation of adhesive molecules renders
microvascular endothelial cells thrombogenic at high shear
stress
Marina Morigi,
Miriam Galbusera,
Elena Binda,
Barbara Imberti,
Sara Gastoldi,
Andrea Remuzzi,
Carla Zoja, and
Giuseppe Remuzzi
From the Mario Negri Institute for Pharmacological
Research, Bergamo, Italy, and the Division of Nephrology and Dialysis,
Azienda Ospedaliera, Ospedali Riuniti di Bergamo, Bergamo, Italy.
Verotoxin-1 (VT-1)-producing Escherichia coli is
the causative agent of postdiarrheal hemolytic uremic
syndrome (D+HUS) of children, which leads to renal and other organ
microvascular thrombosis. Why thrombi form only on arterioles and
capillaries is not known. This study investigated whether VT-1
directly affected endothelial antithrombogenic properties promoting
platelet deposition and thrombus formation on human microvascular
endothelial cell line (HMEC-1) under high shear stress. Human umbilical
vein endothelial cells (HUVECs) were used for comparison as a
large-vessel endothelium. HMEC-1 and HUVECs were pre-exposed for 24 hours to increasing concentrations of VT-1 (2-50 pM) and then perfused
at 60 dynes/cm2 with heparinized human blood prelabeled
with mepacrine. Results showed that VT-1 significantly increased
platelet adhesion and thrombus formation on HMEC-1 in comparison with
unstimulated control cells. An increase in thrombus formation was also
observed on HUVECs exposed to VT-1, but to a remarkably lower extent.
The greater sensitivity of HMEC-1 to the toxin in comparison with HUVECs was at least in part due to a higher expression of VT-1 receptor
(20-fold more) as documented by FACS analysis. The HMEC-1 line had a
comparable susceptibility to the thrombogenic effect of VT-1 as primary
human microvascular cells of the same dermal origin (HDMECs). The
adhesive molecules involved in VT-induced thrombus formation were also
studied. Blocking the binding of von Willebrand factor to platelet
glycoprotein Ib by aurintricarboxylic acid (ATA) or inhibition of
platelet  IIb 3-integrin by chimeric 7E3
Fab resulted in a significant reduction of VT-1-induced thrombus formation, suggesting the involvement of von Willebrand
factor-platelet interaction at high shear stress in this phenomenon.
Functional blockade of endothelial 3-integrin
subunit, vitronectin receptor, P-selectin, and PECAM-1 with
specific antibodies was associated with a significant decrease of the
endothelial area covered by thrombi. Confocal microscopy studies
revealed that VT-1 increased the expression of vitronectin receptor and
P-selectin and redistributed PECAM-1 away from the cell-cell border of
HMEC-1, as well as of HDMECs, thus indicating that the above
endothelial adhesion molecules are directly involved and possibly
determine the effect of VT-1 on enhancing platelet adhesion and
thrombus formation in microvascular endothelium. These results might
help to explain why thrombi in HUS localize in microvessels rather than
in larger ones and provide insights on the molecular events involved in
the process of microvascular thrombosis associated with D+HUS.
 CiteULike  Connotea  Del.icio.us  Digg  Reddit  Technorati What's this?
This article has been cited by other articles:
|
|
|
|
 
C. Zanchi, C. Zoja, M. Morigi, F. Valsecchi, X. Y. Liu, D. Rottoli, M. Locatelli, S. Buelli, A. Pezzotta, P. Mapelli, et al.
Fractalkine and CX3CR1 Mediate Leukocyte Capture by Endothelium in Response to Shiga Toxin
J. Immunol.,
July 15, 2008;
181(2):
1460 - 1469.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Brigotti, D. Carnicelli, E. Ravanelli, A. G. Vara, C. Martinelli, R. R. Alfieri, P. G. Petronini, and P. Sestili
Molecular Damage and Induction of Proinflammatory Cytokines in Human Endothelial Cells Exposed to Shiga Toxin 1, Shiga Toxin 2, and {alpha}-Sarcin
Infect. Immun.,
May 1, 2007;
75(5):
2201 - 2207.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. H. Romer, K. G. Birukov, and J. G.N. Garcia
Focal Adhesions: Paradigm for a Signaling Nexus
Circ. Res.,
March 17, 2006;
98(5):
606 - 616.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. Macconi, M. Bonomelli, A. Benigni, T. Plati, F. Sangalli, L. Longaretti, S. Conti, H. Kawachi, P. Hill, G. Remuzzi, et al.
Pathophysiologic Implications of Reduced Podocyte Number in a Rat Model of Progressive Glomerular Injury
Am. J. Pathol.,
January 1, 2006;
168(1):
42 - 54.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. H. Nolasco, N. A. Turner, A. Bernardo, Z. Tao, T. G. Cleary, J.-f. Dong, and J. L. Moake
Hemolytic uremic syndrome-associated Shiga toxins promote endothelial-cell secretion and impair ADAMTS13 cleavage of unusually large von Willebrand factor multimers
Blood,
December 15, 2005;
106(13):
4199 - 4209.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
F. Guessous, M. Marcinkiewicz, R. Polanowska-Grabowska, S. Kongkhum, D. Heatherly, T. Obrig, and A. R. L. Gear
Shiga Toxin 2 and Lipopolysaccharide Induce Human Microvascular Endothelial Cells To Release Chemokines and Factors That Stimulate Platelet Function
Infect. Immun.,
December 1, 2005;
73(12):
8306 - 8316.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. T. Yamamoto, M. Mizuno, K. Nishikawa, S. Miyazawa, L. Zhang, S. Matsuo, and Y. Natori
Shiga Toxin 1 Causes Direct Renal Injury in Rats
Infect. Immun.,
November 1, 2005;
73(11):
7099 - 7106.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Noris and G. Remuzzi
Hemolytic Uremic Syndrome
J. Am. Soc. Nephrol.,
April 1, 2005;
16(4):
1035 - 1050.
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Soghomonians, T. L. Thirkill, N. F. Mariano, A. I. Barakat, and G. C. Douglas
Effect of Aqueous Tobacco Smoke Extract and Shear Stress on PECAM-1 Expression and Cell Motility in Human Uterine Endothelial Cells
Toxicol. Sci.,
October 1, 2004;
81(2):
408 - 418.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Matussek, J. Lauber, A. Bergau, W. Hansen, M. Rohde, K. E. J. Dittmar, M. Gunzer, M. Mengel, P. Gatzlaff, M. Hartmann, et al.
Molecular and functional analysis of Shiga toxin-induced response patterns in human vascular endothelial cells
Blood,
August 15, 2003;
102(4):
1323 - 1332.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. T. Eitzman, P. F. Bodary, Y. Shen, C. G. Khairallah, S. R. Wild, A. Abe, J. Shaffer-Hartman, and J. A. Shayman
Fabry Disease in Mice Is Associated With Age-Dependent Susceptibility to Vascular Thrombosis
J. Am. Soc. Nephrol.,
February 1, 2003;
14(2):
298 - 302.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. I. Tarr, H.-M. Tsai, W. L. Chandler, C. M. Nzerue, and J. L. Moake
Thrombotic Microangiopathies
N. Engl. J. Med.,
December 26, 2002;
347(26):
2171 - 2173.
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. L. Moake
Thrombotic Microangiopathies
N. Engl. J. Med.,
August 22, 2002;
347(8):
589 - 600.
[Full Text]
[PDF]
|
|
|
|
|
