Basic-alimentary tractProfiling of microdissected gastric epithelial cells reveals a cell type—specific response to Helicobacter pylori infection
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
References (64)
- et al.
A standardized mouse model of Helicobacter pylori infectionintroducing the Sydney strain
Gastroenterology
(1997) - et al.
Identification of a signaling cascade for interleukin-8 production by Helicobacter pylori in human gastric epithelial cells
Biochem Pharmacol
(2002) - et al.
Eicosanoids and the gastrointestinal tract
Gastroenterology
(1995) - et al.
Challenges of single-cell diagnosticsanalysis of gene expression
Trends Mol Med
(2002) - et al.
Portrait of a stem cell
Dev Cell
(2002) - et al.
Molecular characterization of mouse gastric zymogenic cells
J Biol Chem
(2003) Mucosa-related bacteria in the stomach
Lancet
(1984)Host cell signaling in Helicobacter pylori infection
Int J Med Microbiol
(2001)- et al.
Helicobacter pylori-induced prostaglandin E(2) synthesis involves activation of cytosolic phospholipase A(2) in epithelial cells
J Biol Chem
(2001) - et al.
Prostaglandin E2 protects gastric mucosal cells from apoptosis via EP2 and EP4 receptor activation
J Biol Chem
(2003)
Helicobacter pylori infection and the risk for duodenal and gastric ulceration
Ann Intern Med
Living dangerouslyhow Helicobacter pylori survives in the human stomach
Nat Rev Mol Cell Biol
Helicobacter pylori20 years on
Clin Med
Development of a mouse model of Helicobacter pylori infection that mimics human disease
Science
Immune responses of specific-pathogen-free mice to chronic Helicobacter pylori (strain SS1) infection
Infect Immun
Helicobacter pylori-induced mucosal inflammation is Th1 mediated and exacerbated in IL-4, but not IFN-gamma, gene-deficient mice
J Immunol
Helicobacter pylori-associated gastritis in mice is host and strain specific
Infect Immun
MALToma-like lesions in the murine gastric mucosa after long-term infection with Helicobacter felis. A mouse model of Helicobacter pylori-induced gastric lymphoma
Am J Pathol
Molecular characterization of mouse gastric epithelial progenitor cells
Proc Natl Acad Sci U S A
Multipotential stem cells in adult mouse gastric epithelium
Am J Physiol Gastrointest Liver Physiol
Optimization and evaluation of T7 based RNA linear amplification protocols for cDNA microarray analysis
BMC Genomics
The Stanford Microarray Database
Nucleic Acids Res
Cluster analysis and display of genome-wide expression patterns
Proc Natl Acad Sci U S A
Significance analysis of microarrays applied to the ionizing radiation response
Proc Natl Acad Sci U S A
GoMinera resource for biological interpretation of genomic and proteomic data
Genome Biol
DAVIDDatabase for Annotation, Visualization, and Integrated Discovery
Genome Biol
Trefoil peptides and surgical disease
Br J Surg
Selective attraction of monocytes and T lymphocytes of the memory phenotype by cytokine RANTES
Nature
Altered statesinvolvement of phosphorylated CagA in the induction of host cellular growth changes by Helicobacter pylori
Proc Natl Acad Sci U S A
SHP-2 tyrosine phosphatase as an intracellular target of Helicobacter pylori CagA protein
Science
Gene expression profiling using laser capture microdissection
Expert Rev Mol Diagn
mRna expression analysis of tissue sections and single cells
J Neurosci
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Laser microdissection: A promising tool for exploring microorganisms and their interactions with hosts
2017, Journal of Microbiological MethodsCitation Excerpt :Laser microdissection revolutionized the study of host–microorganism interactions at the mucosal surfaces. In several studies, the host responses to commensal or pathogenic bacterial infections in the intestine (Hooper et al., 2001; Jilling et al., 2006; Chowdhury et al., 2007; Preidis et al., 2012; Martins et al., 2013; Vannucci et al., 2013), stomach (Mueller et al., 2003, 2004; Syder et al., 2003; Nomura et al., 2004; Resnick et al., 2006), lung (Zhu et al., 2003; Ly et al., 2007; Brogaard et al., 2015) and bladder (Reigstad et al., 2007) were characterized at the level of gene expression. The host response to fungal infection in the lung has also been described using this method (Xue et al., 2005).
Macrophages promote progression of spasmolytic polypeptide-expressing metaplasia after acute loss of parietal cells
2014, GastroenterologyCitation Excerpt :IFN-γ secreted by T cells drives several different immune processes, including the activation of macrophages and up-regulation of IFN-γ target genes in epithelial cells.13,36 Liu et al used a knockout of Hip1r, a model of parietal cell loss through apoptosis crossed to IFN-γ knockout mice to show reduced metaplasia initially; but over a longer time course, loss of IFN-γ did not impair the development of SPEM.18,37 In contrast, Helicobacter-infected mice overexpressing IFN-γ through the H+/K+-ATPase promoter develop less Helicobacter-associated gastric pathology due to increased epithelial autophagy and decreased parietal cell loss, indicating that IFN-γ might also have a cytoprotective role.35
Integration, visualization and analysis of human interactome
2014, Biochemical and Biophysical Research CommunicationsCitation Excerpt :Although diagnostic and therapeutic advances have increased survival, the prognosis remains low mainly due to GC resistance to treatment [57]. For this reason early treatment for the infection is considered the key to prevent gastric cancer [58] and the use of antibiotics (i.e., tetracycline) as chemoprevention is a highly debated topic [59,60]. Taking into consideration factors that are involved in GC development and the drugs currently used to treat the tumor and H. pylori infection, we use an integrated approach to analyze this tumor.
Impaired gastric gland differentiation in Peutz-Jeghers syndrome
2010, American Journal of PathologyCitation Excerpt :P values indicate where the averaged expression differs significantly from the wild-type expression level (normalized to 1.0 ± 0.0). Also expression of an independent set of 54 chief cell gene identifiers39 not shown in the figure, was significantly (P = 0.0018) decreased in fundic polyps (average expression fold change 0.88 ± 0.36) whereas unaffected Lkb1+/− fundic mucosa had a similar expression signature for this set as the wild-type (1.03 ± 0.16). Despite the similarity between wild-type and unaffected Lkb1+/− mucosa observed in the above gene expression array experiments, manual counting of marker-labeled cells in immunostained tissue sections, pointed to a small increase in foveolar cells and decrease in chief cells and mucopeptic cells (Supplemental figure S3 at http://ajp.amjpathol.org), not fully excluding minor alterations of epithelial composition also in the unaffected Lkb1+/− mucosa.
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.