Altered gene expression in highly purified enterocytes from patients with active coeliac disease.
BACKGROUND
Coeliac disease is a multifactorial inflammatory disorder of the intestine caused by ingestion of gluten in genetically susceptible individuals. Genes within the HLA-DQ locus are considered to contribute some 40% of the genetic influence on this disease. However, information on other disease causing genes is sparse. Since enterocytes are considered to play a central role in coeliac pathology, the aim of this study was to examine gene expression in a highly purified isolate of these cells taken from patients with active disease. Epithelial cells were isolated from duodenal biopsies taken from five coeliac patients with active disease and five non-coeliac control subjects. Contaminating T cells were removed by magnetic sorting. The gene expression profile of the cells was examined using microarray analysis. Validation of significantly altered genes was performed by real-time RT-PCR and immunohistochemistry.


RESULTS
Enterocyte suspensions of high purity (98-99%) were isolated from intestinal biopsies. Of the 3,800 genes investigated, 102 genes were found to have significantly altered expression between coeliac disease patients and controls (p < 0.05). Analysis of these altered genes revealed a number of biological processes that are potentially modified in active coeliac disease. These processes include events likely to contribute to coeliac pathology, such as altered cell proliferation, differentiation, survival, structure and transport.


CONCLUSIONS
This study provides a profile of the molecular changes that occur in the intestinal epithelium of coeliac patients with active disease. Novel candidate genes were revealed which highlight the contribution of the epithelial cell to the pathogenesis of coeliac disease.
Bracken
Suzanne
S
Department of Immunology, St, James's Hospital, Dublin and Trinity College Dublin, Dublin Molecular Medicine Centre, Dublin, Ireland. Suzanne.Bracken@biotrin.ie
Byrne
Greg
G
Kelly
Jacinta
J
Jackson
John
J
Feighery
Conleth
C
D000328:Adult; D016022:Case-Control Studies; D002446:Celiac Disease / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002469:Cell Separation; D017630:Connexins / Q000235:genetics / Q000378:metabolism; D055233:Cornified Envelope Proline-Rich Proteins; D004386:Duodenum / Q000378:metabolism / Q000473:pathology; D020895:Enterocytes / Q000378:metabolism* / Q000473:pathology; D015870:Gene Expression; D006801:Humans; D007150:Immunohistochemistry; D007223:Infant; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D017630:Connexins; D055233:Cornified Envelope Proline-Rich Proteins; D011506:Proteins; C112254:SPRR3 protein, human