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Infection and Immunity, February 2002, p. 685-691, Vol. 70, No. 2
0019-9567/01/$04.00+0     DOI: 70.2.685-691.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

In Vivo Behavior of a Helicobacter pylori SS1 nixA Mutant with Reduced Urease Activity

Kylie J. Nolan,1 David J. McGee,2 Hazel M. Mitchell,1* Tassia Kolesnikow,1 Janette M. Harro,2 Jani O'Rourke,1 John E. Wilson,1 Stephen J. Danon,1 Nathan D. Moss,3 Harry L. T. Mobley,2 and Adrian Lee1

School of Microbiology and Immunology, The University of New South Wales,,1 and ,2 School of Applied Vision Sciences, University of Sydney, Sydney, New South Wales, Australia, and Department of MicrobiologyImmunology, University of Maryland School of Medicine, Baltimore, Maryland 212013

Received 7 May 2001/ Returned for modification 20 August 2001/ Accepted 6 November 2001

Helicobacter pylori mutants devoid of urease activity fail to colonize the gastric mucosa of mice; however, the effect of decreased levels of urease on colonization has not been examined. The nixA gene, required for full urease activity, encodes a cytoplasmic membrane nickel transporter that imports nickel ions and leads to incorporation of nickel ions into apourease. A nixA mutant of the Sydney strain of H. pylori (SS1) was constructed by disruption of the nixA gene with a kanamycin resistance cassette. This mutant retained only half the urease activity of the wild-type (wild-type) SS1 strain. C57BL/6j (n = 75) and BALB/c (n = 75) mice were inoculated independently with the wild-type or the nixA strain. The level and distribution of colonization were assessed by bacterial colony counts and histological grading at 4, 12, and 24 weeks postinfection. Colonization levels of the nixA strain in BALB/c mice were significantly lower compared with SS1 (P = 0.005), while colonization in C57BL/6j mice was similar for both the wild-type and mutant strains. Subtle differences in colonization of the different regions of the stomach, determined by microscopic grading, were observed between wild-type SS1 and the nixA strain in BALB/c mice. On the contrary, when C57BL/6j (n = 35) and BALB/c (n = 35) mice were coinfected with the wild-type and nixA strains simultaneously, the nixA mutant failed to colonize and was outcompeted by the wild-type SS1 strain, which established normal levels of colonization. These results demonstrate the importance of the nixA gene for increasing the fitness of H. pylori for gastric colonization. Since nixA is required for full urease activity, the decreased fitness of the nixA mutant is likely due to reduced urease activity; however, pleiotropic effects of the mutation cannot be completely ruled out.

* Corresponding author. Mailing address: School of Microbiology and Immunology, The University of New South Wales, Sydney, New South Wales 2052, Australia. Phone: 61 (2) 9385 2040. Fax: 61 (2) 9385 1591. E-mail: HMitchell{at}unsw.edu.au.

Editor: V. J. DiRita

Infection and Immunity, February 2002, p. 685-691, Vol. 70, No. 2
0019-9567/01/$04.00+0     DOI: 70.2.685-691.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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