Evidence that verotoxins (Shiga-like toxins) from Escherichia coli bind to P blood group antigens of human erythrocytes in vitro.
The interaction of verotoxins (VTs) with human erythrocytes (RBCs) in vitro was investigated, with particular reference to the role of P blood group glycolipids that are structurally related to the known VT receptors. RBC binding of purified VT1, VT2, VT2c, and VT2e was detected by direct and indirect immunofluorescence. Glycolipids were extracted from defined RBCs, separated by thin-layer chromatography, and assessed for VT binding in an overlay assay by adding toxin and specific antibodies. All VTs bound to P1 phenotype (Pk, P, and P1 antigens) and P2 phenotype (Pk and P antigens) RBCs but not to p phenotype (lacking the Pk, P, and P1 antigens) RBCs. Binding of VT1 and VT2 was approximately 10-fold greater to P1 and the rare Pk2 (Pk antigen but no P1 or P antigen) phenotype cells than to P2 phenotype RBCs, whereas VT2e bound equally well to P1 and P2 phenotype cells. The VT1 and VT2 immunofluorescence results correlated with the detection of P1 and/or increased amounts of Pk (globotriaosylceramide) antigen; VT2e immunofluorescence correlated with the detection of P (globotetraosylceramide) antigen. The Pk band pattern and VT binding observed in the thin-layer chromatogram of human P1 and P phenotype RBC extracts varied from that of human kidney and Pk1 phenotype (Pk and P1 antigens) RBCs. We conclude that each VT binds to human RBCs in vitro by utilizing specific P blood group glycolipids as receptors. On P1 and P phenotype RBCs, the accessibility of the Pk antigen for VTs appeared to be restricted. The occurrence of VT-RBC binding in natural VT-producing Escherichia coli disease and its relevance for the pathophysiology of hemolytic uremic syndrome remain to be established.
Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
M
M
Bitzan
S
S
Richardson
C
C
Huang
B
B
Boyd
M
M
Petric
M A
MA
Karmali
D001427:Bacterial Toxins / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism*; D004926:Escherichia coli / Q000472:pathogenicity*; D005455:Fluorescent Antibody Technique; D006017:Glycolipids / Q000032:analysis / Q000378:metabolism; D006463:Hemolytic-Uremic Syndrome / Q000209:etiology; D006801:Humans; D010127:P Blood-Group System / Q000378:metabolism*; D022622:Shiga Toxin 1; D000519:alpha-Galactosidase / Q000494:pharmacology
D001427:Bacterial Toxins; D006017:Glycolipids; D010127:P Blood-Group System; D022622:Shiga Toxin 1; D000519:alpha-Galactosidase