Gastroenterology

Gastroenterology

Volume 127, Issue 5, November 2004, Pages 1446-1462
Gastroenterology

Basic-alimentary tract
Profiling of microdissected gastric epithelial cells reveals a cell type—specific response to Helicobacter pylori infection

https://doi.org/10.1053/j.gastro.2004.08.054Get rights and content

Background & Aims: Helicobacter pylori colonizes the epithelial lining of the human stomach and is associated with disorders ranging from chronic gastritis to peptic ulcers and gastric cancer. We have explored the transcriptional response of the epithelium globally by applying a whole-genome approach to a murine model of infection. Methods: The 3 major epithelial lineages of the stomach—the parietal, mucus-producing, and chief cells—were harvested from cryosections of infected and uninfected murine stomachs by laser microdissection and subjected to gene expression profiling. The localization and quantity of selected transcripts were verified by in situ hybridization and quantitative real-time reverse-transcription polymerase chain reaction. Results: Each cell type is characterized by a transcriptional signature profile. The parietal cell profile is highly enriched for factors involved in mitochondrial energy generation, whereas the chief cell predominantly expresses digestive enzymes and glycosylation-associated proteins. In contrast, the mucus cell signature is distinguished by an abundance of cell-surface receptors, signaling molecules, and factors involved in antigen presentation. All of these indicate a role in sampling, sensing, and responding to environmental stimuli. In line with this biological function, we measured a strong transcriptional response to Helicobacter pylori infection only in this cell type. The genes that are differentially expressed upon infection are implicated in a proinflammatory and mucosal defense response as well as modulation of angiogenesis, iron availability, and tumor suppression. Conclusions: Laser microdissection—assisted transcriptional profiling is a useful tool to explore the biology of specific cell populations and is sensitive enough to measure the transcriptional response to bacterial infection in vivo.

Section snippets

Bacterial culture and animal infections

Six-week-old female BALB/c mice were infected with the mouse-adapted H pylori strain SS15 for 1, 2, 4, 7, 10, 14, or 28 days. H pylori was grown on solid media on horse blood agar containing 4% Columbia agar base (Oxoid, Basingstoke, Hampshire, UK), 5% defibrinated horse blood (HemoStat Labs), 0.2% β-cyclodextrin, 10 μg/mL vancomycin, 5 μg/mL cefsulodin, 2.5 U/mL polymyxin B, 50 μg/mL cycloheximide, 5 μg/mL trimethoprim, and 8 μg/mL amphotericin (all from Sigma, St. Louis, MO) under

Transcriptional profiling of the three major gastric epithelial lineages

We infected female BALB/c mice with H pylori SS1 for 1, 2, 4, 7, 10, 14, and 28 days and assessed the colonization levels by plating and colony counting (Figure 1). Although there was considerable variation between mice, all animals displayed a clear decrease in bacterial numbers by day 2. By day 4 after infection, bacterial loads had increased, and all animals stably carried a population of H pylori ranging from 2 × 104 to 1 × 105 colony-forming units. We chose the day 2, 7, 14, and 28 mice

Discussion

Microarray-based gene expression profiling is an important tool for understanding and classifying pathophysiological processes because it helps to identify genes and gene pathways not previously associated with particular diseases. Recent technological advances have permitted the analysis of pooled single cells or cell populations that are procured by laser microdissection in a virtually contamination-free manner.24, 25 This approach has been used to purify diseased populations of cells whose

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    Research in the laboratory of S.F. is supported by Grants CA92229 and AI38459 from the National Institutes of Health (NIH). A.M. receives a fellowship (MU1675/1-1) from the Deutsche Forschungsgemeinschaft, Germany, and D.S.M. is funded by NIH Training Grant AI07502-06 and the Damon Runyon Cancer Research Fund.

    A.M. and D.S.M. contributed equally to this work.

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