Eaton Kathryn A, Gilbert Joanne V, Joyce Elizabeth A, Wanken Amy E, Thevenot Tracy, Baker Patrick, Plaut Andrew, Wright Andrew
Department of Veterinary Biosciences, Ohio State University, Columbus, Ohio 43210, USA.
Infect Immun. 2002 Feb;70(2):771-8. doi: 10.1128/IAI.70.2.771-778.2002.
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.
(i)脲酶B阴性的幽门螺杆菌互补后是否能恢复其定植能力;(ii)脲酶是否可作为体内启动子活性的有效报告基因。所用菌株为M6、M6ΔureB,以及M6或M6ΔureB的10种重组衍生物,其中脲酶表达受不同幽门螺杆菌启动子的控制。给小鼠口服接种野生型或其中一种突变菌株,然后测定其定植情况、体内脲酶活性及胃炎程度。在测试的8种M6ΔureB重组体中,有4种能在小鼠体内定植。其中,3种的体外脲酶活性在所有重组体中最高,与不能定植的突变体有显著差异。第4种能定植的重组体中,ureB受cag - 15启动子控制,其体外脲酶活性与不能定植的菌株无显著差异。在体内,4种能定植的转化体和野生型对照的脲酶活性无明显差异。感染M6的小鼠与感染转化体的小鼠在胃炎或上皮病变方面没有差异。这些结果表明,脲酶活性的恢复可使脲酶阴性的幽门螺杆菌恢复定植能力。研究结果还表明,cag - 15在体内上调,正如之前通过证明其在与上皮细胞接触后上调所表明的那样。最后,我们的结果表明,幽门螺杆菌引起的胃炎与总脲酶活性和定植密度无关。