Intestinal Microbiota Composition of Interleukin-10 Deficient C57BL/6J Mice and Susceptibility to Helicobacter hepaticus-Induced Colitis

Abstract

The mouse pathobiont Helicobacter hepaticus can induce typhlocolitis in interleukin-10-deficient mice, and H. hepaticus infection of immunodeficient mice is widely used as a model to study the role of pathogens and commensal bacteria in the pathogenesis of inflammatory bowel disease. C57BL/6J Il10−/− mice kept under specific pathogen-free conditions in two different facilities (MHH and MIT), displayed strong differences with respect to their susceptibilities to H. hepaticus-induced intestinal pathology. Mice at MIT developed robust typhlocolitis after infection with H. hepaticus, while mice at MHH developed no significant pathology after infection with the same H. hepaticus strain. We hypothesized that the intestinal microbiota might be responsible for these differences and therefore performed high resolution analysis of the intestinal microbiota composition in uninfected mice from the two facilities by deep sequencing of partial 16S rRNA amplicons. The microbiota composition differed markedly between mice from both facilities. Significant differences were also detected between two groups of MHH mice born in different years. Of the 119 operational taxonomic units (OTUs) that occurred in at least half the cecum or colon samples of at least one mouse group, 24 were only found in MIT mice, and another 13 OTUs could only be found in MHH samples. While most of the MHH-specific OTUs could only be identified to class or family level, the MIT-specific set contained OTUs identified to genus or species level, including the opportunistic pathogen, Bilophila wadsworthia. The susceptibility to H. hepaticus-induced colitis differed considerably between Il10−/− mice originating from the two institutions. This was associated with significant differences in microbiota composition, highlighting the importance of characterizing the intestinal microbiome when studying murine models of IBD.

TRIF Signaling Drives Homeostatic Intestinal Epithelial Antimicrobial Peptide Expression

“…The microbiota was analyzed as detailed by Yang et al (30) with modifications. Bacterial DNA was isolated using the QIAamp DNA stool kit (Qiagen, Hilden) according to manufacturer's instructions.…”

“…After subsampling to an equal number of sequences per sample, rarefaction curves were generated with mothur. The distribution of OTUs among the samples was visualized using the R script heat map (36) as described by Yang et al (30). The R package vegan (37) was used for principal coordinates analyses (PCoA), using the "capscale" command in combination with the Bray-Curtis dissimilarity index, and for fitting the factors mouse genotype, litter, and mouse sex onto the PCoA ordination using "envfit."…”

The gastric microbial community,Helicobacter pyloricolonization, and disease

“…For example, Yang et al found that H. hepaticus could induce colitis in IL10 −/− mice bred at only one of two breeding facilities, and this trait was associated with a distinct intestinal microbiota in uninfected mice. 39 A complex gut microbiota has also been associated with enhanced clearance of H. felis, another gastric helicobacter. 40 However, studies performed by us and others that have looked for similar vendor or antibiotic effects on H. pylori colonization have met with negative results.…”

Actual Usage and Quality of Experimental Colitis Models in Preclinical Efficacy Testing

“…C57BL/6J IL-10 2/2 mice held in different facilities and kept under specific pathogen-free conditions showed a significant difference in susceptibility to an H. hepaticus infection seemingly associated with their microbiota composition. 30 Another study shows that different laboratory diets influence susceptibility to DSS colitis. Each of the diets cause a different microbiota composition, which results in varying susceptibilities.…”