Rapid genetic analysis of Helicobacter pylori gastric mucosal colonization in suckling mice.
Previously described animal models for Helicobacter pylori infection have been limited by cumbersome host requirements (e.g., germ-free conditions or unusual species) or are applicable to only special subsets of H. pylori strains (e.g., fresh clinical isolates or animal-adapted derivatives). Here, we report that 5- to 6-day-old outbred CD-1 (ICR) suckling mice support 24-h colonization of all H. pylori strains tested (SS1, 26695 SmR-1, 43504 SmR-1, and G27 SmR-1), including lab-passaged strains that cannot be adapted for colonization of adult animals. Total colony-forming units (cfu) recovered from infection with lab-passaged strains did not differ from those with mouse-adapted SS1. We also tested this model's ability to detect colonization defects in strains carrying mutations in known virulence genes by coinfecting with wild-type H. pylori and measuring differential recovery. This competition assay identified colonization defects in several classes of known attenuated mutants, including those defective in acid resistance (ureA), metabolism (frdA), motility (motB), and chemotaxis (cheY). A mutant defective in copA (copper transporting P-type ATPase) is nonattenuated in adult and infant mice. Possibly because of the limited duration of infection, our model did not identify defects in vacuolating cytotoxin (vacA) or gamma-glutamyltranspeptidase (ggt) as attenuating, in contrast to results from other animal models. We also identified a new virulence gene (HP0507) encoding a conserved hypothetical protein, which is important for colonization in our model. The suckling mouse model offers a rapid method to identify colonization defects in any H. pylori strain and may have utility as a new tool for studying immunity to primary infection.
Guo
Betty P
BP
Department of Microbiology and Molecular Genetics, Harvard Medical School, 200 Longwood Avenue, Armenise 408, Boston, MA 02115.
Mekalanos
John J
JJ
D000375:Aging; D000595:Amino Acid Sequence; D000818:Animals; D000833:Animals, Suckling; D017931:DNA Primers; D005753:Gastric Mucosa / Q000382:microbiology*; D016480:Helicobacter pylori / Q000254:growth & development / Q000302:isolation & purification / Q000472:pathogenicity*; D006801:Humans; D051379:Mice; D008813:Mice, Inbred ICR; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers