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. 2017 Sep 13;7(1):11450.
doi: 10.1038/s41598-017-11734-8.

Butyrate-producing bacteria supplemented in vitro to Crohn's disease patient microbiota increased butyrate production and enhanced intestinal epithelial barrier integrity

Annelies Geirnaert  1   2 , Marta Calatayud  1 , Charlotte Grootaert  3 , Debby Laukens  4 , Sarah Devriese  4 , Guy Smagghe  5 , Martine De Vos  4 , Nico Boon  1 , Tom Van de Wiele  6
Affiliations

Butyrate-producing bacteria supplemented in vitro to Crohn's disease patient microbiota increased butyrate production and enhanced intestinal epithelial barrier integrity

Annelies Geirnaert et al. Sci Rep. .

Abstract

The management of the dysbiosed gut microbiota in inflammatory bowel diseases (IBD) is gaining more attention as a novel target to control this disease. Probiotic treatment with butyrate-producing bacteria has therapeutic potential since these bacteria are depleted in IBD patients and butyrate has beneficial effects on epithelial barrier function and overall gut health. However, studies assessing the effect of probiotic supplementation on microbe-microbe and host-microbe interactions are rare. In this study, butyrate-producing bacteria (three mono-species and one multispecies mix) were supplemented to the fecal microbial communities of ten Crohn's disease (CD) patients in an in vitro system simulating the mucus- and lumen-associated microbiota. Effects of supplementation in short-chain fatty acid levels, bacterial colonization of mucus environment and intestinal epithelial barrier function were evaluated. Treatment with F. prausnitzii and the mix of six butyrate-producers significantly increased the butyrate production by 5-11 mol%, and colonization capacity in mucus- and lumen-associated CD microbiota. Treatments with B. pullicaecorum 25-3T and the mix of six butyrate-producers improved epithelial barrier integrity in vitro. This study provides proof-of-concept data for the therapeutic potential of butyrate-producing bacteria in CD and supports the future preclinical development of a probiotic product containing butyrate-producing species.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Microbial parameters of non-treated microbiota from CD patients with active disease (A; red) (n = 5) versus in remission (R; blue) (n = 5). SCFA production after 18 h, 42 h and 65 h of incubation (A): Boxplot of absolute SCFA concentration (downward diagonal pattern) and relative SCFA concentration (filled boxes). Black lines within boxplot represent median values and whiskers indicate minimum and maximum value. The concentration of Ruminococcaceae species (Clostridium cluster IV) (B) and Lachnospiraceae species (Clostridium cluster XIVa) (C) in fecal samples and lumen and mucus samples of non-treated microbiota after 65 h of incubation. Black lines within data plot represent mean values. Significant differences on 0.05 level between average value of active and remission samples are indicated with an asterisk.
Figure 2
Figure 2
Treatment with butyrate-producers results in higher butyrate production. Δ Mol% represents the difference in SCFA levels (acetate, propionate and butyrate) between samples treated with butyrate-producers and corresponding non-treated samples. Average values and SEM are plotted for samples of active CD (A) and CD in remission (R) after 18 h, 42 h and 65 h of treatment. Treatments are indicated with letter codes: BP 25-3: B. pullicaecorum 25-3; BP 1-20: B. pullicaecorum 1.20; FP: F. prausnitzii; MIX: mix of B. pullicaecorum 25-3, F. prausnitzii, R. intestinalis, R. hominis, E. hallii, A. caccae. Significant differences at 0.05 level between treatments are indicated with different letter codes, the averages of bars with the same letter in their letter code are not significant different from each other.
Figure 3
Figure 3
Colonization success of different treatments in mucus and lumen samples of CD microbiota (A: active; R: remission) at end of incubation. Based on group-specific DGGE profiles by comparing the relative intensity of the band classes of the different butyrate-producing species used in treatments between non-treated and treated samples. Colonization success of the supplemented butyrate-producers was defined if the relative intensity in the treated sample was higher as in the non-treated sample. In case of the MIX, there was a colonization success defined if the relative intensity of the band class of at least one species was higher in treated sample. Treatments are indicated with letter codes: BP 25-3: B. pullicaecorum 25-3; BP 1-20: B. pullicaecorum 1.20; FP: F. prausnitzii; MIX: mix of B. pullicaecorum 25-3, F. prausnitzii, R. intestinalis, R. hominis, E. hallii, A. caccae.
Figure 4
Figure 4
Overview of the colonization successes of the different members of the MIX in mucus- and lumen-associated butyrate-producing bacterial communities of active CD (A, n = 5) or CD in remission (R, n = 5) at the end of incubation. For each sample the number and type of butyrate-producer that colonized the microbiota is given.
Figure 5
Figure 5
Epithelial barrier integrity at day 4 and day 9 of differentiation of Caco-2 epithelial cells treated with 1/10 (v/v) microbial supernatant of incubated CD microbiota, which were treated or not (no probiotic) with B. pullicaecorum 25-3T (BP25-3) or a mix of six butyrate-producers (MIX). Data of BP25-3 and MIX are expressed as % of the ‘no probiotic’. Average TEER and Papp LY values with Stdev are shown (n ≥ 3) of the six assays (3 active (A), 3 remissive (R) patients) performed. Significant differences with corresponding ‘no probiotic’ sample are indicated with an asterisk.
Figure 6
Figure 6
Effect of butyrate-spiking on epithelial barrier integrity. Epithelial barrier integrity at day 4 and day 9 of differentiation of Caco-2 epithelial cells treated with 1/10 (v/v) microbial supernatant of incubated CD microbiota (boxes with downward diagonal pattern), which were treated or not (no probiotic) with B. pullicaecorum 25-3T (BP25-3) or a mix of six butyrate-producers (MIX). In addition, 1/10 (v/v) diluted supernatant samples were spiked with butyrate (final concentration of 2 mM) (dark grey boxes). Data of spiked ‘no probiotic’, BP25-3 and MIX are expressed as % of the non-spiked ‘no probiotic’. Average TEER and Papp LY values with stdev are shown (n ≥ 3) of the two assays (active patient 5, A5, and patient in remission 4, R4). Significant differences with corresponding ‘no probiotic’ sample are indicated with an asterisk.
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References

    1. Danese S, Fiocchi C. Etiopathogenesis of inflammatory bowel diseases. World J Gastroenterol. 2006;12:4807–4812. - PMC - PubMed
    1. Zhang Y-Z, Li Y-Y. Inflammatory bowel disease: Pathogenesis. World journal of gastroenterology: WJG. 2014;20:91. doi: 10.3748/wjg.v20.i1.91. - DOI - PMC - PubMed
    1. Torres J, Danese S, Colombel JF. New therapeutic avenues in ulcerative colitis: thinking out of the box. Gut. 2013;62:1642–1652. doi: 10.1136/gutjnl-2012-303959. - DOI - PubMed
    1. Coskun, M. Intestinal epithelium in inflammatory bowel disease. Frontiers in Medicine1, 10.3389/fmed.2014.00024 (2014). - PMC - PubMed
    1. Bernstein CN. Treatment of IBD: Where We Are and Where We Are Going. Am J Gastroenterol. 2015;110:114–126. doi: 10.1038/ajg.2014.357. - DOI - PubMed

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