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Infect. Immun., 05 1995, 1681-1687, Vol 63, No. 5
SA Sharma, MK Tummuru, GG Miller and MJ Blaser
Gastric infection with Helicobacter pylori activates a mucosal inflammatory
response by mononuclear cells and neutrophils that includes expression of
cytokines interleukin-1 beta (IL-1 beta), IL-6, tumor necrosis factor
alpha, and IL-8. In this study, we analyzed the IL-8 response of human
gastric cancer cell lines (Kato III, AGS, and MKN28) to H. pylori infection
in vitro. IL-8 mRNA expression was detected by reverse transcription-PCR
amplification of RNA extracted from epithelial cells after incubation with
different H. pylori wild- type and mutant strains, and IL-8 secretion was
measured by an enzyme- linked immunosorbent assay. Exposure to viable H.
pylori induced IL-8 mRNA and protein synthesis in all three gastric cell
lines but not in nongastric epithelial cell lines. Heat-killed H. pylori
and a crude cytotoxin preparation did not induce significant IL-8
secretion. IL-8 mRNA peaked between 2 and 4 h postinfection, and IL-8
protein production was maximal 24 h postinfection. Exposure of gastric
carcinoma cells to other gastrointestinal bacteria, such as Pseudomonas
aeruginosa, Campylobacter jejuni, and Escherichia coli, but not
Campylobacter fetus, induced IL-8 synthesis. Wild-type strains that
expressed the vacuolating cytotoxin (Tox+) and a cytotoxin-associated gene
(cagA) product (CagA+) induced significantly more IL-8 than did CagA- Tox-
strains. However, there was no decrease in IL-8 induction by isogenic
mutants of CagA-, Tox-, or Cag- Tox- strains or by a mutant lacking the
urease subunits. These results indicate that exposure to H. pylori and
other gram-negative organisms that do not colonize the gastric mucosa
induces IL-8 production by gastric carcinoma cells in vitro. Although the
CagA+ Tox+ phenotype of H. pylori is associated with enhanced IL-8
production by gastric cell lines, other bacterial constituents are clearly
essential.
Copyright © 1995, American Society for Microbiology
Interleukin-8 response of gastric epithelial cell lines to Helicobacter pylori stimulation in vitro
Department of Medicine, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2605, USA.
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