The Toll-like receptors TLR2 and TLR4 do not affect the intestinal microbiota composition in mice.

The interaction between intestinal epithelial cells and microbes is partly mediated by Toll-like receptors (TLRs). Sensing of Gram-positive and Gram-negative bacteria by TLR2 and TLR4, respectively, can result in immune system activation and in an exclusion of bacteria from the intestine. To test the impact of these TLRs on bacterial composition, germ-free TLR2/TLR4 double-knock out mice and the corresponding C57BL/10ScSn wild-type mice where associated with fecal bacteria from one single donor mouse. In addition, C3H/HeOuJ and BALB/c mice were used in this study. Fecal bacteria were monitored over 13 weeks with denaturing-gradient gel electrophoresis (DGGE). Colonic bacteria were enumerated by fluorescent in situ hybridization (FISH) and short-chain fatty acids (SCFA) were measured in caecal samples. No effect of the TLRs on intestinal microbiota composition and SCFA concentrations was observed. However, the microbiota composition as reflected by DGGE band patterns differed between C3H and BALB/c mice on the one hand and C57BL/10 mice on the other hand. Corresponding differences between the mouse strains were also observed in cecal propionic, valeric and i-valeric acid concentrations. No differences between the animals were observed in the numbers of bacteria detected by FISH. We conclude that genetic traits but not TLR2 and TLR4 have an impact on the intestinal microbiota composition.

[1]  U. Göbel,et al.  Exacerbation of murine ileitis by Toll-like receptor 4 mediated sensing of lipopolysaccharide from commensal Escherichia coli , 2007, Gut.

[2]  E. Mardis,et al.  An obesity-associated gut microbiome with increased capacity for energy harvest , 2006, Nature.

[3]  H. Glatt,et al.  Mutagenicity of arbutin in mammalian cells after activation by human intestinal bacteria. , 2006, Food and chemical toxicology : an international journal published for the British Industrial Biological Research Association.

[4]  R. Sartor.  Mechanisms of Disease: pathogenesis of Crohn's disease and ulcerative colitis , 2006, Nature Clinical Practice Gastroenterology &Hepatology.

[5]  J. Gordon,et al.  A humanized gnotobiotic mouse model of host-archaeal-bacterial mutualism. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[6]  P. Rutgeerts,et al.  Toll‐like receptor‐1, ‐2, and ‐6 polymorphisms influence disease extension in inflammatory bowel diseases , 2006, Inflammatory bowel diseases.

[7]  R. Bibiloni,et al.  Analysis of the large bowel microbiota of colitic mice using PCR/DGGE , 2005, Letters in applied microbiology.

[8]  H. Flint,et al.  pH and Peptide Supply Can Radically Alter Bacterial Populations and Short-Chain Fatty Acid Ratios within Microbial Communities from the Human Colon , 2005, Applied and Environmental Microbiology.

[9]  Vaahtovuo Jussi,et al.  Comparison of cellular fatty acid profiles of the microbiota in different gut regions of BALB/c and C57BL/6J mice , 2005, Antonie van Leeuwenhoek.

[10]  E. Purdom,et al.  Diversity of the Human Intestinal Microbial Flora , 2005, Science.

[11]  M. Blaut,et al.  An improved method for the automated enumeration of fluorescently labelled bacteria in human faeces. , 2005, Journal of microbiological methods.

[12]  E. Cario.  BACTERIAL INTERACTIONS WITH CELLS OF THE INTESTINAL MUCOSA: TOLL-LIKE RECEPTORS AND NOD2 , 2005, Gut.

[13]  G. Macfarlane,et al.  Mucosal bacteria in ulcerative colitis , 2005, British Journal of Nutrition.

[14]  Ting Wang,et al.  The gut microbiota as an environmental factor that regulates fat storage. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[15]  K. Van Steen,et al.  Deficient host-bacteria interactions in inflammatory bowel disease? The toll-like receptor (TLR)-4 Asp299gly polymorphism is associated with Crohn’s disease and ulcerative colitis , 2004, Gut.

[16]  H. Harmsen,et al.  Analysis of 16S libraries of mouse gastrointestinal microflora reveals a large new group of mouse intestinal bacteria. , 2002, Microbiology.

[17]  H. Harmsen,et al.  Extensive Set of 16S rRNA-Based Probes for Detection of Bacteria in Human Feces , 2002, Applied and Environmental Microbiology.

[18]  P. Toivanen,et al.  Study of murine faecal microflora by cellular fatty acid analysis; effect of age and mouse strain , 2001, Antonie van Leeuwenhoek.

[19]  Willem M. de Vos,et al.  The Host Genotype Affects the Bacterial Community in the Human Gastrointestinal Tract , 2001 .

[20]  J. Gordon,et al.  Commensal Host-Bacterial Relationships in the Gut , 2001, Science.

[21]  P. Toivanen,et al.  Influence of Major Histocompatibility Complex on Bacterial Composition of Fecal Flora , 2001, Infection and Immunity.

[22]  J. Klein,et al.  The HLA system. Second of two parts. , 2000 .

[23]  J. Doré,et al.  Quantification of Bacterial Groups within Human Fecal Flora by Oligonucleotide Probe Hybridization , 2000, Applied and Environmental Microbiology.

[24]  S. Akira,et al.  Differential roles of TLR2 and TLR4 in recognition of gram-negative and gram-positive bacterial cell wall components. , 1999, Immunity.

[25]  H. Harmsen,et al.  A 16S rRNA-targeted Probe for Detection of Lactobacilli and Enterococci in Faecal Samples by Fluorescent In Situ Hybridization , 1999 .

[26]  E. Zoetendal,et al.  Temperature Gradient Gel Electrophoresis Analysis of 16S rRNA from Human Fecal Samples Reveals Stable and Host-Specific Communities of Active Bacteria , 1998, Applied and Environmental Microbiology.

[27]  Gerwin C. Raangs,et al.  Variations of Bacterial Populations in Human Feces Measured by Fluorescent In Situ Hybridization with Group-Specific 16S rRNA-Targeted Oligonucleotide Probes , 1998, Applied and Environmental Microbiology.

[28]  R. Amann,et al.  Sequence heterogeneities of genes encoding 16S rRNAs in Paenibacillus polymyxa detected by temperature gradient gel electrophoresis , 1996, Journal of bacteriology.

[29]  R Amann,et al.  Application of a suite of 16S rRNA-specific oligonucleotide probes designed to investigate bacteria of the phylum cytophaga-flavobacter-bacteroides in the natural environment. , 1996, Microbiology.

[30]  M. Wilkinson,et al.  Quantitative fluorescence in situ hybridization of Bifidobacterium spp. with genus-specific 16S rRNA-targeted probes and its application in fecal samples , 1995, Applied and environmental microbiology.

[31]  K. Schleifer,et al.  Phylogenetic identification and in situ detection of individual microbial cells without cultivation. , 1995, Microbiological reviews.

[32]  A. Simpson,et al.  Rapid silver staining and recovery of PCR products separated on polyacrylamide gels. , 1994, BioTechniques.

[33]  J. Doré,et al.  Enumeration of Bacteroides species in human faeces by fluorescent in situ hybridisation combined with flow cytometry using 16S rRNA probes. , 2003, Systematic and applied microbiology.

[34]  J. Gordon,et al.  How host-microbial interactions shape the nutrient environment of the mammalian intestine. , 2002, Annual review of nutrition.

[35]  J. Klein,et al.  The HLA system. Second of two parts. , 2000, The New England journal of medicine.

[36]  W. Scheppach,et al.  Faecal short-chain fatty acids after colonic surgery. , 1989, European journal of clinical nutrition.

[37]  Daniel B. Oerther,et al.  The oligonucleotide probe database , 1996, Applied and environmental microbiology.