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Infection and Immunity, November 2001, p. 6970-6980, Vol. 69, No. 11
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.11.6970-6980.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
cDNA Array Analysis of cag Pathogenicity
Island-Associated Helicobacter pylori Epithelial Cell
Response Genes
Joanne M.
Cox,1
Christopher L.
Clayton,2
Toshihiko
Tomita,1
Don M.
Wallace,2
Philip A.
Robinson,1 and
Jean E.
Crabtree1,*
Molecular Medicine Unit, St. James's
University Hospital, Leeds LS9 7TF,1 and
Genomics Unit, Glaxo Smith Kline Research and Development,
Stevenage, Hertfordshire SG1 2NY,2 United
Kingdom
Received 15 March 2001/Returned for modification 25 May
2001/Accepted 20 June 2001
Helicobacter pylori strains containing the
cag pathogenicity island (PAI) induce NF-
B activation
and interleukin-8 secretion in gastric epithelial cells. The aim of
this study was to investigate changes in epithelial gene expression
induced by cag PAI-positive and -negative strains of
H. pylori using high-density cDNA array hybridization
technology. Radio-labeled cDNA prepared from H. pylori-infected Kato 3 gastric epithelial cells was hybridized to
high-density cDNA arrays to identify changes in epithelial gene
expression compared to noninfected controls. In vivo expression of
selected, differentially expressed genes was examined by reverse transcription-PCR analysis of H. pylori-positive and
-negative gastric mucosa. Screening of ca. 57,800 cDNAs identified 208 known genes and 48 novel genes and/or expressed sequence tags of
unknown function to be differentially expressed in Kato 3 cells
following H. pylori infection. Marked differences in gene
expression profiles were observed following cag
PAI-positive and cag PAI-negative infection with 15 novel
cDNAs and 92 known genes being differentially expressed. H. pylori was found to change the expression of genes encoding
growth factors and cytokine/chemokines and their receptors, apoptosis
proteins, transcription factors and metalloprotease-disintegrin proteins (ADAMs), and tissue inhibitors of metalloproteinases. Gastric
differential expression of selected known genes (amphiregulin and ADAM
10) and a novel gene (HPYR1) was confirmed in vivo in patients with H. pylori infection. Confirmation of the in
vivo expression of selected genes demonstrates the usefulness of this approach for investigating pathogen-induced changes in host gene expression.
*
Corresponding author. Mailing address: Level 7, Clinical Sciences Building, St. James's University Hospital, Leeds LS9
7TF, United Kingdom. Phone: 44-113-2065267. Fax: 44-113-2429722. E-mail: MSJJC{at}stjames.leeds.ac.uk.
Infection and Immunity, November 2001, p. 6970-6980, Vol. 69, No. 11
0019-9567/01/$04.00+0 DOI: 10.1128/IAI.69.11.6970-6980.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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