Increased adherence and expression of virulence genes in a lineage of Escherichia coli O157:H7 commonly associated with human infections.
BACKGROUND
Enterohemorrhagic Escherichia coli (EHEC) O157:H7, a food and waterborne pathogen, can be classified into nine phylogenetically distinct lineages, as determined by single nucleotide polymorphism genotyping. One lineage (clade 8) was found to be associated with hemolytic uremic syndrome (HUS), which can lead to kidney failure and death in some cases, particularly young children. Another lineage (clade 2) differs considerably in gene content and is phylogenetically distinct from clade 8, but caused significantly fewer cases of HUS in a prior study. Little is known, however, about how these two lineages vary with regard to phenotypic traits important for disease pathogenesis and in the expression of shared virulence genes.


RESULTS
Here, we quantified the level of adherence to and invasion of MAC-T bovine epithelial cells, and examined the transcriptomes of 24 EHEC O157:H7 strains with varying Shiga toxin profiles from two common lineages. Adherence to epithelial cells was >2-fold higher for EHEC O157:H7 strains belonging to clade 8 versus clade 2, while no difference in invasiveness was observed between the two lineages. Whole-genome 70-mer oligo microarrays, which probe for 6088 genes from O157:H7 Sakai, O157:H7 EDL 933, pO157, and K12 MG1655, detected significant differential expression between clades in 604 genes following co-incubation with epithelial cells for 30 min; 186 of the 604 genes had a >1.5 fold change difference. Relative to clade 2, clade 8 strains showed upregulation of major virulence genes, including 29 of the 41 locus of enterocyte effacement (LEE) pathogenicity island genes, which are critical for adherence, as well as Shiga toxin genes and pO157 plasmid-encoded virulence genes. Differences in expression of 16 genes that encode colonization factors, toxins, and regulators were confirmed by qRT-PCR, which revealed a greater magnitude of change than microarrays.


CONCLUSIONS
These findings demonstrate that the EHEC O157:H7 lineage associated with HUS expresses higher levels of virulence genes and has an enhanced ability to attach to epithelial cells relative to another common lineage.
Abu-Ali
Galeb S
GS
Microbial Evolution Laboratory, National Food Safety & Toxicology Center, Michigan State University, East Lansing, Michigan, United States of America.
Ouellette
Lindsey M
LM
Henderson
Scott T
ST
Lacher
David W
DW
Riordan
James T
JT
Whittam
Thomas S
TS
Manning
Shannon D
SD
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000382:microbiology; D004927:Escherichia coli Infections / Q000235:genetics / Q000382:microbiology; D019453:Escherichia coli O157 / Q000235:genetics* / Q000472:pathogenicity*; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D016680:Genome, Bacterial; D006463:Hemolytic-Uremic Syndrome / Q000235:genetics* / Q000382:microbiology; D006801:Humans; D014774:Virulence / Q000235:genetics