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Infection and Immunity, August 2001, p. 4759-4766, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4759-4766.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Assessment of Helicobacter pylori Gene Expression within Mouse and Human Gastric Mucosae by Real-Time Reverse Transcriptase PCR

Bachra Rokbi,1,* Delphine Seguin,1 Bruno Guy,1 Véronique Mazarin,1 Emmanuel Vidor,1 François Mion,2 Michel Cadoz,1 and Marie-José Quentin-Millet1

Aventis Pasteur1 and Fédération des Spécialités Digestives, Hôpital Edouard Herriot,2 Lyon, France

Received 28 March 2001/Accepted 2 May 2001

Despite increasing knowledge on the biology of Helicobacter pylori, little is known about the expression pattern of its genome during infection. While mouse models of infection have been widely used for the screening of protective antigens, the reliability of the mouse model for gene expression analysis has not been assessed. In an attempt to address this question, we have developed a quantitative reverse transcriptase PCR (RT-PCR) that allowed the detection of minute amounts of mRNA within the gastric mucosa. The expression of four genes, 16S rRNA, ureA (encoding urease A subunit), katA (catalase), and alpA (an adhesin), was monitored during the course of a 6-month infection of mice and in biopsy samples from of 15 infected humans. We found that the selected genes were all expressed within both mouse and human infected mucosae. Moreover, the relative abundance of transcripts was the same (16S rRNA > ureA > katA > alpA), in the two models. Finally, results obtained with the mouse model suggest a negative effect of bacterial burden on the number of transcripts of each gene expressed per CFU (P < 0.05 for 16S rRNA, alpA, and katA). Overall, this study demonstrates that real-time RT-PCR is a powerful tool for the detection and quantification of H. pylori gene expression within the gastric mucosa and strongly indicates that mice experimentally infected with H. pylori provide a valuable model for the analysis of bacterial gene expression during infection.

* Corresponding author. Mailing address: Aventis Pasteur, Campus Mérieux, 1541 avenue Marcel Mérieux, 69280 Marcy L'Etoile. Phone: (33)04.37.37.36.05. Fax: (33)04.37.37.31.89. E-mail: Bachra.Rokbi{at}aventis.com.

Infection and Immunity, August 2001, p. 4759-4766, Vol. 69, No. 8
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.8.4759-4766.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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