Platelet activation by Shiga toxin and circulatory factors as a pathogenetic mechanism in the hemolytic uremic syndrome.
Thrombocytopenia caused by platelet consumption in thrombi is a major manifestation of hemolytic uremic syndrome (HUS) associated with Shiga toxin (Stx) producing Escherichia coli. Platelets have glycosphingolipid receptors capable of binding Stx, but a direct interaction between the toxin and platelets, leading to platelet activation, has not been reported. In this study, it is shown that Stx1 and its B (binding) subunit (Stx1B), at 10 pg/mL to 10 ng/mL, bound to platelets. Toxin was internalized in platelets within 2 hours. This led to increased platelet aggregation, as demonstrated by confocal microscopy. Preincubation of Stx1B with anti-Stx1 antibody inhibited this reaction. Stx1 induced morphologic changes in platelets seen on scanning electron microscopy. In the presence of platelets and tumor necrosis factor-pretreated human umbilical vein endothelial cells (HUVEC), Stx1 and Stx1B induced the binding of platelets to the endothelial cell membrane and were present at this binding site. Incubation of Stx1 and Stx1B with whole blood increased fibrinogen binding to platelets detected by flow cytometry. Fibrinogen binding was partially inhibited by preincubation with anti-Stx1. Stx1 increased platelet retention measured in a glass bead assay. In addition, plasma from 17 patients with HUS, taken during the acute phase of the disease, increased the retention of normal platelets and normalized after recovery. Taken together, the results of this investigation show that Stx1, Stx1B, and a factor or factors in the plasma of patients with HUS activate platelets. The presence of Stx1 at the binding site of platelets to HUVEC suggests that Stx may be directly involved in the prothrombotic state seen in HUS.
Karpman
D
D
Department of Pediatrics, Lund University, Lund, Sweden. diana.karpman@skane.se
Papadopoulou
D
D
Nilsson
K
K
Sjögren
A C
AC
Mikaelsson
C
C
Lethagen
S
S
D000293:Adolescent; D001792:Blood Platelets / Q000378:metabolism / Q000648:ultrastructure; D002648:Child; D002675:Child, Preschool; D004730:Endothelium, Vascular / Q000502:physiology; D004926:Escherichia coli / Q000378:metabolism; D004927:Escherichia coli Infections / Q000097:blood; D005260:Female; D005340:Fibrinogen / Q000378:metabolism; D006463:Hemolytic-Uremic Syndrome / Q000097:blood* / Q000382:microbiology; D006801:Humans; D007223:Infant; D008297:Male; D015539:Platelet Activation / Q000187:drug effects*; D022621:Shiga Toxin / Q000378:metabolism / Q000494:pharmacology*; D022622:Shiga Toxin 1 / Q000378:metabolism / Q000494:pharmacology; D022641:Shiga Toxin 2 / Q000378:metabolism / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D014471:Umbilical Veins
D022622:Shiga Toxin 1; D022641:Shiga Toxin 2; D014409:Tumor Necrosis Factor-alpha; D022621:Shiga Toxin; D005340:Fibrinogen