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Infection and Immunity, March 2001, p. 1697-1703, Vol. 69, No. 3
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.3.1697-1703.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Helicobacter pylori Pore-Forming Cytolysin Orthologue TlyA Possesses In Vitro Hemolytic Activity and Has a Role in Colonization of the Gastric Mucosa

M. Celeste Martino,1,dagger Richard A. Stabler,1 Zun W. Zhang,2 Michael J. G. Farthing,2 Brendan W. Wren,1 and Nick Dorrell1,*

Pathogen Molecular Biology and Biochemistry Unit, Department of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine,1 and Digestive Diseases Research Centre, St. Bartholomew's and the Royal London School of Medicine and Dentistry, Whitechapel,2 London, United Kingdom

Received 18 October 2000/Returned for modification 16 November 2000/Accepted 13 December 2000

Hemolysins have been found to possess a variety of functions in bacteria, including a role in virulence. Helicobacter pylori demonstrates hemolytic activity when cultured on unlysed blood agar plates which is increased under iron-limiting conditions. However, the role of an H. pylori hemolysin in virulence is unclear. Scrutiny of the H. pylori 26695 genome sequence suggests the presence of at least two distinct hemolysins, HP1086 and HP1490, in this strain. Previous studies have shown that the in vitro hemolytic activity of H. pylori is reduced when it is coincubated with dextran 5000, suggesting the presence of a pore-forming cytolysin. HP1086 has homology to pore-forming cytolysins (TlyA) from other bacterial species, and the introduction of the cloned H. pylori tlyA gene into a nonhemolytic Escherichia coli strain conferred hemolytic activity. An H. pylori tlyA defined mutant showed reduced in vitro hemolytic activity, which appears to be due to pore formation, as the hemolytic activity of the wild-type strain is reduced to the same level as the tlyA mutant by the addition of dextran 5000. The mutant also showed reduced adhesion to human gastric adenocarcinoma cells and failed to colonize the gastric mucosa of mice. These data clearly suggest a role in virulence for H. pylori TlyA, contrary to the suggestion that hemolytic activity is an in vitro phenomenon for this pathogen.

* Corresponding author. Mailing address: Pathogen Molecular Biology and Biochemistry Unit, Department of Infectious and Tropical Diseases, London School of Hygiene & Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom. Phone: 44 (0)20 7612 7853. Fax: 44 (0)20 7637 4314. E-mail: nick.dorrell{at}lshtm.ac.uk.

dagger Present address: Department of Immunology, Immunobiological Research Institute of Siena (IRIS), Chiron Vaccines, Siena, Italy.

Infection and Immunity, March 2001, p. 1697-1703, Vol. 69, No. 3
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.3.1697-1703.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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