Abstract
Background. Little is known about levels of expression of Helicobacter pylori genes in the human host. We therefore developed a quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) assay to measure transcript profiles of H. pylori in the human stomach. Methods. In vivo expression of 16 genes on the cag pathogenicity island and of 18 putative virulence genes was quantitated by isolation of total RNA directly from infected human gastric mucosa. The results were compared with in vitro expression determined from H. pylori cells grown in culture. Results. The highest levels of expression were found for cag1 and cag25 and for genes, such as urease and catalase, that may be important for bacterial homeostasis in the relatively hostile environment of the gastric mucosa. Transcript abundance, relative to 16S rRNA, was lower in vivo than in vitro, which suggests that H. pylori cells are in stationary phase in the gastric environment. This was particularly apparent for cagA. Since CagA is arguably of unique importance, in terms of interaction with the host, tight control of its in vivo expression might be particularly important. Conclusions. qRT-PCR is a powerful tool to measure gene expression in human or animal tissue that contains minute amounts of microbial mRNA, and the results reflect on the physiology of the pathogen in its natural host.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 946-956 |
| Number of pages | 11 |
| Journal | Journal of Infectious Diseases |
| Volume | 190 |
| Issue number | 5 |
| DOIs | |
| State | Published - Sep 1 2004 |
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ASJC Scopus subject areas
- Public Health, Environmental and Occupational Health
- Immunology
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Transcription profile of Helicobacter pylori in the human stomach reflects its physiology in vivo. / Boonjakuakul, Jenni K.; Syvanen, Michael; Suryaprasad, Arun; Bowlus, Christopher; Solnick, Jay V.
In: Journal of Infectious Diseases, Vol. 190, No. 5, 01.09.2004, p. 946-956.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Transcription profile of Helicobacter pylori in the human stomach reflects its physiology in vivo
AU - Boonjakuakul, Jenni K.
AU - Syvanen, Michael
AU - Suryaprasad, Arun
AU - Bowlus, Christopher
AU - Solnick, Jay V
PY - 2004/9/1
Y1 - 2004/9/1
N2 - Background. Little is known about levels of expression of Helicobacter pylori genes in the human host. We therefore developed a quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) assay to measure transcript profiles of H. pylori in the human stomach. Methods. In vivo expression of 16 genes on the cag pathogenicity island and of 18 putative virulence genes was quantitated by isolation of total RNA directly from infected human gastric mucosa. The results were compared with in vitro expression determined from H. pylori cells grown in culture. Results. The highest levels of expression were found for cag1 and cag25 and for genes, such as urease and catalase, that may be important for bacterial homeostasis in the relatively hostile environment of the gastric mucosa. Transcript abundance, relative to 16S rRNA, was lower in vivo than in vitro, which suggests that H. pylori cells are in stationary phase in the gastric environment. This was particularly apparent for cagA. Since CagA is arguably of unique importance, in terms of interaction with the host, tight control of its in vivo expression might be particularly important. Conclusions. qRT-PCR is a powerful tool to measure gene expression in human or animal tissue that contains minute amounts of microbial mRNA, and the results reflect on the physiology of the pathogen in its natural host.
AB - Background. Little is known about levels of expression of Helicobacter pylori genes in the human host. We therefore developed a quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) assay to measure transcript profiles of H. pylori in the human stomach. Methods. In vivo expression of 16 genes on the cag pathogenicity island and of 18 putative virulence genes was quantitated by isolation of total RNA directly from infected human gastric mucosa. The results were compared with in vitro expression determined from H. pylori cells grown in culture. Results. The highest levels of expression were found for cag1 and cag25 and for genes, such as urease and catalase, that may be important for bacterial homeostasis in the relatively hostile environment of the gastric mucosa. Transcript abundance, relative to 16S rRNA, was lower in vivo than in vitro, which suggests that H. pylori cells are in stationary phase in the gastric environment. This was particularly apparent for cagA. Since CagA is arguably of unique importance, in terms of interaction with the host, tight control of its in vivo expression might be particularly important. Conclusions. qRT-PCR is a powerful tool to measure gene expression in human or animal tissue that contains minute amounts of microbial mRNA, and the results reflect on the physiology of the pathogen in its natural host.
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UR - http://www.scopus.com/inward/citedby.url?scp=4344597845&partnerID=8YFLogxK
U2 - 10.1086/423142
DO - 10.1086/423142
M3 - Article
C2 - 15295700
AN - SCOPUS:4344597845
VL - 190
SP - 946
EP - 956
JO - Journal of Infectious Diseases
JF - Journal of Infectious Diseases
SN - 0022-1899
IS - 5
ER -