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Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness.

Pickard JM, Maurice CF, Kinnebrew MA, Abt MC, Schenten D, Golovkina TV, Bogatyrev SR, Ismagilov RF, Pamer EG, Turnbaugh PJ, Chervonsky AV - Nature (2014)

Bottom Line: Fucose affects the expression of microbial metabolic pathways and reduces the expression of bacterial virulence genes.It also improves host tolerance of the mild pathogen Citrobacter rodentium.Thus, rapid IEC fucosylation appears to be a protective mechanism that utilizes the host's resources to maintain host-microbial interactions during pathogen-induced stress.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Committee on Immunology, The University of Chicago, Chicago, Illinois 60637, USA.

ABSTRACT
Systemic infection induces conserved physiological responses that include both resistance and 'tolerance of infection' mechanisms. Temporary anorexia associated with an infection is often beneficial, reallocating energy from food foraging towards resistance to infection or depriving pathogens of nutrients. However, it imposes a stress on intestinal commensals, as they also experience reduced substrate availability; this affects host fitness owing to the loss of caloric intake and colonization resistance (protection from additional infections). We hypothesized that the host might utilize internal resources to support the gut microbiota during the acute phase of the disease. Here we show that systemic exposure to Toll-like receptor (TLR) ligands causes rapid α(1,2)-fucosylation of small intestine epithelial cells (IECs) in mice, which requires the sensing of TLR agonists, as well as the production of interleukin (IL)-23 by dendritic cells, activation of innate lymphoid cells and expression of fucosyltransferase 2 (Fut2) by IL-22-stimulated IECs. Fucosylated proteins are shed into the lumen and fucose is liberated and metabolized by the gut microbiota, as shown by reporter bacteria and community-wide analysis of microbial gene expression. Fucose affects the expression of microbial metabolic pathways and reduces the expression of bacterial virulence genes. It also improves host tolerance of the mild pathogen Citrobacter rodentium. Thus, rapid IEC fucosylation appears to be a protective mechanism that utilizes the host's resources to maintain host-microbial interactions during pathogen-induced stress.

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Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPSa, Silver-stained SDS-PAGE of UEA-1-precipitated SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; *P<0.05 [one-way ANOVA with Bonferroni post-test (d), two-tailed Student's t test (e)]; representative of 3 independent experiments. f,fucA and fucP gene expression relative to housekeeping gene rpoA (Quantitative RT-PCR) in E. coli tested as in d. *P<0.05 by ANOVA with Bonferroni's post-test. g, Stable relative abundance of bacterial phyla across treatment groups and genotypes, as indicated by shotgun sequencing of community DNA. Phyla with a mean RPKM (reads per kilobase per million mapped reads) >40,000 are shown including Actinobacteria (purple), Bacteroidetes (red), Firmicutes (blue), Fusobacteria (green), Proteobacteria (orange), and Tenericutes (yellow). 16S rRNA gene sequencing confirms these observations. Extended Data Fig. 7 shows 16S rRNA gene sequencing results. h, Differentially expressed KEGG orthologous groups following LPS treatment (paired glm edgeR analysis; q<0.05, >2-fold change; see Supplemental Information Table 2 for complete list). i, Increased gut microbial expression of fucose permease (fucP; KO2429) in Fut2-sufficient mice (mean±s.e.m.; *P<0.01, Mann-Whitney test).
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Figure 3: Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPSa, Silver-stained SDS-PAGE of UEA-1-precipitated SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; *P<0.05 [one-way ANOVA with Bonferroni post-test (d), two-tailed Student's t test (e)]; representative of 3 independent experiments. f,fucA and fucP gene expression relative to housekeeping gene rpoA (Quantitative RT-PCR) in E. coli tested as in d. *P<0.05 by ANOVA with Bonferroni's post-test. g, Stable relative abundance of bacterial phyla across treatment groups and genotypes, as indicated by shotgun sequencing of community DNA. Phyla with a mean RPKM (reads per kilobase per million mapped reads) >40,000 are shown including Actinobacteria (purple), Bacteroidetes (red), Firmicutes (blue), Fusobacteria (green), Proteobacteria (orange), and Tenericutes (yellow). 16S rRNA gene sequencing confirms these observations. Extended Data Fig. 7 shows 16S rRNA gene sequencing results. h, Differentially expressed KEGG orthologous groups following LPS treatment (paired glm edgeR analysis; q<0.05, >2-fold change; see Supplemental Information Table 2 for complete list). i, Increased gut microbial expression of fucose permease (fucP; KO2429) in Fut2-sufficient mice (mean±s.e.m.; *P<0.01, Mann-Whitney test).

Mentions: To understand the reason for slow weight recovery in Fut2−/− mice, we tested whether fucosylation affected the function of host IEC proteins identified by direct sequencing as secreted mucins and digestive enzymes (Fig. 3a). Fucosylation did not change activity of several enzymes (Fig. 3b). Thus, changes in enzymatic activity are unlikely to explain the slow weight recovery in Fut2−/− mice, although the role of less abundant fucosylated proteins cannot be excluded.


Rapid fucosylation of intestinal epithelium sustains host-commensal symbiosis in sickness.

Pickard JM, Maurice CF, Kinnebrew MA, Abt MC, Schenten D, Golovkina TV, Bogatyrev SR, Ismagilov RF, Pamer EG, Turnbaugh PJ, Chervonsky AV - Nature (2014)

Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPSa, Silver-stained SDS-PAGE of UEA-1-precipitated SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; *P<0.05 [one-way ANOVA with Bonferroni post-test (d), two-tailed Student's t test (e)]; representative of 3 independent experiments. f,fucA and fucP gene expression relative to housekeeping gene rpoA (Quantitative RT-PCR) in E. coli tested as in d. *P<0.05 by ANOVA with Bonferroni's post-test. g, Stable relative abundance of bacterial phyla across treatment groups and genotypes, as indicated by shotgun sequencing of community DNA. Phyla with a mean RPKM (reads per kilobase per million mapped reads) >40,000 are shown including Actinobacteria (purple), Bacteroidetes (red), Firmicutes (blue), Fusobacteria (green), Proteobacteria (orange), and Tenericutes (yellow). 16S rRNA gene sequencing confirms these observations. Extended Data Fig. 7 shows 16S rRNA gene sequencing results. h, Differentially expressed KEGG orthologous groups following LPS treatment (paired glm edgeR analysis; q<0.05, >2-fold change; see Supplemental Information Table 2 for complete list). i, Increased gut microbial expression of fucose permease (fucP; KO2429) in Fut2-sufficient mice (mean±s.e.m.; *P<0.01, Mann-Whitney test).
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Figure 3: Commensals utilize fucose detached from proteins fucosylated by Fut2 upon systemic challenge with LPSa, Silver-stained SDS-PAGE of UEA-1-precipitated SI IEC protein from control or LPS-treated mouse. b, Ratios of digestive enzymes activities in SI IECs of LPS-treated to untreated Fut2-sufficient (black bars) or Fut2-deficient (open bars) mice two days after LPS injection. Mean±s.e.m. of 4 combined experiments, 4 mice/group. c, SDS-PAGE of intestinal contents blotted on nitrocellulose and stained with UEA-1-peroxidase complexes. s, SI; c, cecum; f, feces; abx, antibiotic treated mice; DFJ, deoxyfuconojirimycin. d, e, Fucose-sensitive GFP reporter expression in gnotobiotic mice colonized with the indicated strains (d, ) or SPF mice (e). Dots are values for individual bacteria, lines are means, n=120; *P<0.05 [one-way ANOVA with Bonferroni post-test (d), two-tailed Student's t test (e)]; representative of 3 independent experiments. f,fucA and fucP gene expression relative to housekeeping gene rpoA (Quantitative RT-PCR) in E. coli tested as in d. *P<0.05 by ANOVA with Bonferroni's post-test. g, Stable relative abundance of bacterial phyla across treatment groups and genotypes, as indicated by shotgun sequencing of community DNA. Phyla with a mean RPKM (reads per kilobase per million mapped reads) >40,000 are shown including Actinobacteria (purple), Bacteroidetes (red), Firmicutes (blue), Fusobacteria (green), Proteobacteria (orange), and Tenericutes (yellow). 16S rRNA gene sequencing confirms these observations. Extended Data Fig. 7 shows 16S rRNA gene sequencing results. h, Differentially expressed KEGG orthologous groups following LPS treatment (paired glm edgeR analysis; q<0.05, >2-fold change; see Supplemental Information Table 2 for complete list). i, Increased gut microbial expression of fucose permease (fucP; KO2429) in Fut2-sufficient mice (mean±s.e.m.; *P<0.01, Mann-Whitney test).
Mentions: To understand the reason for slow weight recovery in Fut2−/− mice, we tested whether fucosylation affected the function of host IEC proteins identified by direct sequencing as secreted mucins and digestive enzymes (Fig. 3a). Fucosylation did not change activity of several enzymes (Fig. 3b). Thus, changes in enzymatic activity are unlikely to explain the slow weight recovery in Fut2−/− mice, although the role of less abundant fucosylated proteins cannot be excluded.

Bottom Line: Fucose affects the expression of microbial metabolic pathways and reduces the expression of bacterial virulence genes.It also improves host tolerance of the mild pathogen Citrobacter rodentium.Thus, rapid IEC fucosylation appears to be a protective mechanism that utilizes the host's resources to maintain host-microbial interactions during pathogen-induced stress.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Committee on Immunology, The University of Chicago, Chicago, Illinois 60637, USA.

ABSTRACT
Systemic infection induces conserved physiological responses that include both resistance and 'tolerance of infection' mechanisms. Temporary anorexia associated with an infection is often beneficial, reallocating energy from food foraging towards resistance to infection or depriving pathogens of nutrients. However, it imposes a stress on intestinal commensals, as they also experience reduced substrate availability; this affects host fitness owing to the loss of caloric intake and colonization resistance (protection from additional infections). We hypothesized that the host might utilize internal resources to support the gut microbiota during the acute phase of the disease. Here we show that systemic exposure to Toll-like receptor (TLR) ligands causes rapid α(1,2)-fucosylation of small intestine epithelial cells (IECs) in mice, which requires the sensing of TLR agonists, as well as the production of interleukin (IL)-23 by dendritic cells, activation of innate lymphoid cells and expression of fucosyltransferase 2 (Fut2) by IL-22-stimulated IECs. Fucosylated proteins are shed into the lumen and fucose is liberated and metabolized by the gut microbiota, as shown by reporter bacteria and community-wide analysis of microbial gene expression. Fucose affects the expression of microbial metabolic pathways and reduces the expression of bacterial virulence genes. It also improves host tolerance of the mild pathogen Citrobacter rodentium. Thus, rapid IEC fucosylation appears to be a protective mechanism that utilizes the host's resources to maintain host-microbial interactions during pathogen-induced stress.

Show MeSH
Related in: MedlinePlus