Verotoxin 2 enhances adherence of enterohemorrhagic Escherichia coli O157:H7 to intestinal epithelial cells and expression of {beta}1-integrin by IPEC-J2 cells.
Verotoxin (VT) has been implicated in the promotion of adherence to and colonization of intestinal epithelial cells by enterohemorrhagic Escherichia coli (EHEC) O157:H7. The present study investigated the effect of VT2 on the adherence of EHEC O157:H7 strain 86-24 to porcine jejunal (IPEC-J2), human colon (CaCo-2), and human laryngeal carcinoma (HEp-2) cell lines and on the expression in IPEC-J2 cells of synthases for beta1-integrin and nucleolin, both of which are implicated in bacterial adherence. The effect on expression of globotriaosylceramide (Gb3) synthase, the receptor for VT, was also examined. Data were obtained by adherence assays and quantitative reverse transcriptase PCR, using EHEC O157 strain 86-24, a vt2 deletion mutant, a vt2 phage-negative strain, and complemented mutants in which the vt2 gene was restored. Compared with the adherence of the parent and complemented mutant strains, the vt2-negative strains adhered significantly less to all three types of cells. Adherence of the wild-type EHEC strain to IPEC-J2 cells was accompanied by increased expression of beta1-integrin, nucleolin, and Gb3 synthase. IPEC-J2 cells in association with wild-type EHEC O157:H7 or the complemented mutants expressed higher levels of beta1-integrin than did cells in association with the vt2-negative strains or with no bacteria. Expression of nucleolin was decreased by association with the vt2-negative mutant, but complementation failed to restore wild-type expression. The data indicate that VT2 plays a role in the adherence of EHEC O157:H7 to intestinal epithelial cells, possibly by increasing the expression of the host receptor beta1-integrin.
Liu
Bianfang
B
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, Ontario, Canada N1G 2W1.
Yin
Xianhua
X
Feng
Yanni
Y
Chambers
James R
JR
Guo
Aiguang
A
Gong
Joshua
J
Zhu
Jing
J
Gyles
Carlton L
CL
D000818:Animals; D001422:Bacterial Adhesion / Q000187:drug effects*; D002460:Cell Line; D045744:Cell Line, Tumor; D003106:Colon / Q000166:cytology / Q000382:microbiology; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology*; D019453:Escherichia coli O157 / Q000378:metabolism / Q000502:physiology*; D005700:Galactosyltransferases / Q000378:metabolism; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D007583:Jejunum / Q000166:cytology / Q000382:microbiology*; D010750:Phosphoproteins / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D022641:Shiga Toxin 2 / Q000378:metabolism* / Q000494:pharmacology; D013552:Swine; D000097796:Nucleolin
D005700:Galactosyltransferases; D019012:Integrin beta1; D010750:Phosphoproteins; D016601:RNA-Binding Proteins; D022641:Shiga Toxin 2; D000097796:Nucleolin; C045097:UDP-galactose-lactosylceramide alpha 1-4-galactosyltransferase
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