The hemorrhagic coli pilus (HCP) of Escherichia coli O157:H7 is an inducer of proinflammatory cytokine secretion in intestinal epithelial cells.
BACKGROUND
Enterohemorrhagic Escherichia coli (EHEC) O157:H7, the causative agent of hemorrhagic colitis and the hemolytic uremic syndrome (HUS), produces long bundles of type IV pili (TFP) called hemorrhagic coli pili (HCP). HCP are capable of mediating several phenomena associated with pathogenicity: i) adherence to human and bovine epithelial cells; ii) invasion of epithelial cells; iii) hemagglutination of rabbit erythrocytes; iv) biofilm formation; v) twitching motility; and vi) specific binding to laminin and fibronectin. HCP are composed of a 19 kDa pilin subunit (HcpA) encoded by the hcpA chromosomal gene (called prepilin peptidase-dependent gene [ppdD] in E. coli K-12).


RESULTS
In this study we investigated the potential role of HCP of E. coli O157:H7 strain EDL933 in activating the release of pro- and anti-inflammatory cytokines from a variety of host epithelial cells. We found that purified HCP and a recombinant HcpA protein induced significant release of IL-8 and TNF-alpha, from cultured polarized intestinal cells (T84 and HT-29 cells) and non-intestinal HeLa cells. Levels of proinflammatory IL-8 and TNF-alpha, but not IL-2, IL6, or IL-10 cytokines, were increased in the presence of HCP and recombinant HcpA after 6 h of incubation with >or=50 ng/ml of protein, suggesting that stimulation of IL-8 and TNF-alpha are dose and time-dependent. In addition, we also demonstrated that flagella are potent inducers of cytokine production. Furthermore, MAPK activation kinetics studies showed that EHEC induces p38 phosphorylation under HCP-producing conditions, and ERK1/2 and JNK activation was detectable after 3 h of EHEC infection. HT-29 cells were stimulated with epidermal growth factor stimulation of HT-29 cells for 30 min leading to activation of three MAPKs.


CONCLUSIONS
The HcpA pilin monomer of the HCP produced by EHEC O157:H7 is a potent inducer of IL-8 and TNF-alpha release, an event which could play a significant role in the pathogenesis of hemorrhagic colitis caused by this pathogen.
Ledesma
Maria A
MA
Laboratorio de Bacteriología Intestinal, Departamento de Infectología, Hospital Infantil de México Federico Gómez, México Distrito Federal, Mexico.
Ochoa
Sara A
SA
Cruz
Ariadnna
A
Rocha-Ramírez
Luz M
LM
Mas-Oliva
Jaime
J
Eslava
Carlos A
CA
Girón
Jorge A
JA
Xicohtencatl-Cortes
Juan
J
D000818:Animals; D000906:Antibodies / Q000276:immunology; D002417:Cattle; D045744:Cell Line, Tumor; D016764:Cell Polarity; D016207:Cytokines / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D019453:Escherichia coli O157 / Q000378:metabolism* / Q000502:physiology; D036781:Fimbriae Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology*; D005407:Flagella / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D007249:Inflammation / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D007413:Intestinal Mucosa / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects; D016328:NF-kappa B / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D013997:Time Factors; D018808:Transcription Factor AP-1 / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D000906:Antibodies; D016207:Cytokines; D016209:Interleukin-8; D016328:NF-kappa B; D021122:Protein Subunits; D018808:Transcription Factor AP-1; D014409:Tumor Necrosis Factor-alpha; D036781:Fimbriae Proteins