In vivo complementation of ureB restores the ability of Helicobacter pylori to colonize.
The objective of this study was to determine (i) if complementation of ureB-negative Helicobacter pylori restores colonization and (ii) if urease is a useful reporter for promoter activity in vivo. Strains used were M6, M6DeltaureB, and 10 recombinant derivatives of M6 or M6DeltaureB in which urease expression was under the control of different H. pylori promoters. Mice were orally inoculated with either the wild type or one of the mutant strains, and colonization, in vivo urease activity, and extent of gastritis were determined. Of eight M6DeltaureB recombinants tested, four colonized mice. Of those, three had the highest in vitro urease activity of any of the recombinants, significantly different from that of the noncolonizing mutants. The fourth colonizing recombinant, with ureB under control of the cag-15 promoter, had in vitro urease activity which did not differ significantly from the noncolonizing strains. In vivo, urease activities of the four colonizing transformants and the wild-type control were indistinguishable. There were no differences in gastritis or epithelial lesions between mice infected with M6 and those infected with the transformants. These results demonstrate that recovery of urease activity can restore colonizing ability to urease-negative H. pylori. They also suggest that cag-15 is upregulated in vivo, as was previously suggested by demonstrating that it is upregulated upon contact with epithelial cells. Finally, our results suggest that total urease activity and colonization density do not contribute to gastritis due to H. pylori.
Eaton
Kathryn A
KA
Department of Veterinary Biosciences, Ohio State University, Columbus, Ohio 43210, USA. eaton.1@osu.edu
Gilbert
Joanne V
JV
Joyce
Elizabeth A
EA
Wanken
Amy E
AE
Thevenot
Tracy
T
Baker
Patrick
P
Plaut
Andrew
A
Wright
Andrew
A
D000818:Animals; D005260:Female; D005756:Gastritis / Q000382:microbiology / Q000473:pathology; D005816:Genetic Complementation Test; D016481:Helicobacter Infections / Q000382:microbiology / Q000473:pathology; D016480:Helicobacter pylori / Q000201:enzymology* / Q000254:growth & development; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D016296:Mutagenesis; D014510:Urease / Q000235:genetics / Q000378:metabolism*
D014510:Urease