Enterohemorrhagic Escherichia coli O157:H7 gene expression profiling in response to growth in the presence of host epithelia.
BACKGROUND
The pathogenesis of enterohemorrhagic Escherichia coli (EHEC) O157:H7 infection is attributed to virulence factors encoded on multiple pathogenicity islands. Previous studies have shown that EHEC O157:H7 modulates host cell signal transduction cascades, independent of toxins and rearrangement of the cytoskeleton. However, the virulence factors and mechanisms responsible for EHEC-mediated subversion of signal transduction remain to be determined. Therefore, the purpose of this study was to first identify differentially regulated genes in response to EHEC O157:H7 grown in the presence of epithelial cells, compared to growth in the absence of epithelial cells (that is, growth in minimal essential tissue culture medium alone, minimal essential tissue culture medium in the presence of 5% CO(2), and Penassay broth alone) and, second, to identify EHEC virulence factors responsible for pathogen modulation of host cell signal transduction.


RESULTS
Overnight cultures of EHEC O157:H7 were incubated for 6 hr at 37 degrees C in the presence or absence of confluent epithelial (HEp-2) cells. Total RNA was then extracted and used for microarray analyses (Affymetrix E. coli Genome 2.0 gene chips). Relative to bacteria grown in each of the other conditions, EHEC O157:H7 cultured in the presence of cultured epithelial cells displayed a distinct gene-expression profile. A 2.0-fold increase in the expression of 71 genes and a 2.0-fold decrease in expression of 60 other genes were identified in EHEC O157:H7 grown in the presence of epithelial cells, compared to bacteria grown in media alone.


CONCLUSIONS
Microarray analyses and gene deletion identified a protease on O-island 50, gene Z1787, as a potential virulence factor responsible for mediating EHEC inhibition of the interferon (IFN)-gamma-Jak1,2-STAT-1 signal transduction cascade. Up-regulated genes provide novel targets for use in developing strategies to interrupt the infectious process.
Jandu
Narveen
N
Department of Pathology, Stanford University School of Medicine, Stanford, CA, USA.
Ho
Nathan K L
NK
Donato
Kevin A
KA
Karmali
Mohamed A
MA
Mascarenhas
Mariola
M
Duffy
Simon P
SP
Tailor
Chetankumar
C
Sherman
Philip M
PM
D002460:Cell Line; D004847:Epithelial Cells / Q000382:microbiology; D019453:Escherichia coli O157 / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity; D020869:Gene Expression Profiling; D005798:Genes, Bacterial; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D014774:Virulence / Q000235:genetics