Induction of a chemoattractant transcriptional response by a Campylobacter jejuni boiled cell extract in colonocytes.
BACKGROUND
Campylobacter jejuni, the commonest cause of bacterial diarrhoea worldwide, can also induce colonic inflammation. To understand how a previously identified heat stable component contributes to pro-inflammatory responses we used microarray and real-time quantitative PCR to investigate the transcriptional response to a boiled cell extract of Campylobacter jejuni NCTC 11168.


RESULTS
RNA was extracted from the human colonocyte line HCA-7 (clone 29) after incubation for 6 hours with Campylobacter jejuni boiled cell extract and was used to probe the Affymetrix Human Genome U133A array. Genes differentially affected by Campylobacter jejuni boiled cell extract were identified using the Significance Score algorithm of the Bioconductor software suite and further analyzed using the Ingenuity Pathway Analysis program. The chemokines CCL20, CXCL3, CXCL2, Interleukin 8, CXCL1 and CXCL6 comprised 6 of the 10 most highly up-regulated genes, all with Significance Scores > or = 10. Members of the Tumor Necrosis Factor alpha/Nuclear Factor-kappaB super-family were also significantly up-regulated and involved in the most significantly regulated signalling pathways (Death receptor, Interleukin 6, Interleukin 10, Toll like receptor, Peroxisome Proliferator Activated Receptor-gamma and apoptosis). Ingenuity Pathway Analysis also identified the most affected functional gene networks such as cell movement, gene expression and cell death. In contrast, down-regulated genes were predominantly concerned with structural and metabolic functions.


CONCLUSIONS
A boiled cell extract of Campylobacter jejuni has components that can directly switch the phenotype of colonic epithelial cells from one of resting metabolism to a pro-inflammatory one, particularly characterized by increased expression of genes for leukocyte chemoattractant molecules.
Mellits
Kenneth H
KH
Division of Food Sciences, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, Leicestershire, LE12 5RD, UK. ken.mellits@nottingham.ac.uk
Connerton
Ian F
IF
Loughlin
Michael F
MF
Clarke
Peter
P
Smith
Julie
J
Dillon
Eleanor
E
Connerton
Phillippa L
PL
Mulholland
Francis
F
Hawkey
Christopher J
CJ
D016123:Campylobacter jejuni / Q000737:chemistry* / Q000276:immunology*; D002460:Cell Line; D018925:Chemokines / Q000096:biosynthesis; D002630:Chemotactic Factors / Q000276:immunology* / Q000302:isolation & purification; D003106:Colon / Q000166:cytology / Q000276:immunology*; D015536:Down-Regulation; D004847:Epithelial Cells / Q000276:immunology*; D020869:Gene Expression Profiling; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000276:immunology; D015854:Up-Regulation
D018925:Chemokines; D002630:Chemotactic Factors