Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets.
A mutant strain of Helicobacter pylori with weak urease activity was created by using N-methyl-N'-nitro-N-nitrosoguanidine. The urease activity of the mutant (0.036 +/- 0.009 nmol of urea per micrograms of bacterial protein per min) was 0.4% of that of the parental strain (8.20 +/- 2.30 nmol of urea per micrograms of bacterial protein per min). The mutant was otherwise indistinguishable from the parental strain. Both demonstrated prominent catalase and oxidase activities, and both produced vacuolating cytotoxin. Restriction endonuclease and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns and ultrastructure were identical for the two strains. The mutant was fully motile, as evaluated by spreading in soft agar and by direct microscopic examination. Growth rate and colony size and morphology were identical for the mutant and parental strains. Seventeen gnotobiotic piglets were challenged with either the mutant or the parental strain and sacrificed 3 or 21 days after challenge. Gastric tissue was examined histologically and cultured for H. pylori. Of seven piglets challenged with the parental strain, all became infected. H. pylori was not recovered from any of 10 piglets challenged with the urease-negative strain. Lymphofollicular gastritis was present in all seven piglets challenged with the parental strain but in none of the piglets challenged with the urease-negative strain. These results suggest that prominent urease activity is essential for colonization by H. pylori.
Eaton
K A
KA
Ohio State University, Columbus 43210.
Brooks
C L
CL
Morgan
D R
DR
Krakowka
S
S
D000818:Animals; D004269:DNA, Bacterial / Q000235:genetics; D005856:Germ-Free Life; D016480:Helicobacter pylori / Q000472:pathogenicity* / Q000648:ultrastructure; D008854:Microscopy, Electron; D016296:Mutagenesis; D015183:Restriction Mapping; D013552:Swine; D014510:Urease / Q000235:genetics / Q000378:metabolism*
D004269:DNA, Bacterial; D014510:Urease