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. 2017 Feb 1:8:114.
doi: 10.3389/fmicb.2017.00114. eCollection 2017.

Microbial Anti-Inflammatory Molecule (MAM) from Faecalibacterium prausnitzii Shows a Protective Effect on DNBS and DSS-Induced Colitis Model in Mice through Inhibition of NF-κB Pathway (VSports最新版本)

Natalia M Breyner  1 , Cristophe Michon  1 , Cassiana S de Sousa  2 , Priscilla B Vilas Boas  2 , Florian Chain  1 , Vasco A Azevedo  3 , Philippe Langella  1 , Jean M Chatel  1
Affiliations

Microbial Anti-Inflammatory Molecule (MAM) from Faecalibacterium prausnitzii Shows a Protective Effect on DNBS and DSS-Induced Colitis Model in Mice through Inhibition of NF-κB Pathway

Natalia M Breyner (V体育官网) et al. Front Microbiol. .

Abstract

Faecalibacterium prausnitzii and its supernatant showed protective effects in different chemically-induced colitis models in mice. Recently, we described 7 peptides found in the F. prausnitzii supernatant, all belonging to a protein called Microbial Anti-inflammatory Molecule (MAM). These peptides were able to inhibit NF-κB pathway in vitro and showed anti-inflammatory properties in vivo in a DiNitroBenzene Sulfate (DNBS)-induced colitis model. In this current proof we tested MAM effect on NF-κB pathway in vivo, using a transgenic model of mice producing luciferase under the control of NF-κB promoter. Moreover, we tested this protein on Dextran Sodium Sulfate (DSS)-induced colitis in mice. To study the effect of MAM we orally administered to the mice a Lactococcus lactis strain carrying a plasmid containing the cDNA of MAM under the control of a eukaryotic promoter VSports手机版. L. lactis delivered plasmids in epithelial cells of the intestinal membrane allowing thus the production of MAM directly by host. We showed that MAM administration inhibits NF-κB pathway in vivo. We confirmed the anti-inflammatory properties of MAM in DNBS-induced colitis but also in DSS model. In DSS model MAM was able to inhibit Th1 and Th17 immune response while in DNBS model MAM reduced Th1, Th2, and Th17 immune response and increased TGFβ production. .

Keywords: Faecalibacterium prausnitzii; Lactococcus lactis; MAM (Microbial Anti-inflammatory Molecule); NF-κB; colitis models; inflammation V体育安卓版. .

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Figures

Figure 1
Figure 1
Inhibition of NF-κB pathway in vivo after LL-pILMAM administration. NF-κB luciferase mice, transgenic mice expressing luciferase under the control of NF-κB, were orally administered with PBS, LL-pILEMPTY or LL-pILMAM 7 days before DNBS intrarectal injection (D0) and until sacrifice (D4). NF-κB activation was monitored by luminescence in vivo on whole animal using IVIS Spectrum. ROI (Regions of interest) measurements were used to determinate how many photons are radiating from the source.
Figure 2
Figure 2
Cytokines secreted by reactivated lymphocytes from MLN in a DNBS-induced colitis model. NF-κB luciferase mice were orally administered with PBS, LL-pILEMPTY or LL-pILMAM 7 days before DNBS intrarectal injection (D0) and until sacrifice (D4). At D4 MLN were withdrawn and isolated lymphocytes reactivated by anti-CD3/anti-CD28 antibodies. Cytokine concentration in medium was monitored 48 h after reactivation by ELISA. *P < 0.05.
Figure 3
Figure 3
Cytokines produced by colon tissue in DNBS-induced colitis model. NF-κB luciferase mice were orally administered with PBS, LL-pILEMPTY or LL-pILMAM 7 days before DNBS intrarectal injection (D0) and until sacrifice (D4). At D4 colon was withdrawn and proteins extracted. Cytokine concentration in protein extracts was monitored by ELISA. *P < 0.05, **P < 0.01.
Figure 4
Figure 4
Effect of MAM on macroscopic scores in a DSS-induced colitis model. Mice were orally administered with LL-pILMAM, LL-pILEMPTY or PBS 7 days before and during colitis induction. Colitis was induced by adding DSS in drinking water during 7 days (D0-D7). Then DSS was removed from drinking water and mice allowed to recover during 5 days (D7-D12). Weight (A) and DAI (B) including bleeding (C) were monitored daily. *P < 0.05.
Figure 5
Figure 5
Cytokines secreted by reactivated lymphocytes from MLN in DSS-induced colitis model. Mice were orally administered with LL-pILMAM, LL-pILEMPTY or PBS 7 days before and during colitis induction. Colitis was induced by adding DSS in drinking water during 7 days (D0-D7). Then DSS was removed from drinking water and mice allowed to recover during 5 days (D7-D12). At D12 mice were killed, MLN withdrawn, cells were isolated and lymphocytes reactivated with antiCD3/anti-CD28 antibodies. Cytokine concentration in medium was monitored 48 h after reactivation by ELISA. *P < 0.05, **P < 0.01.
Figure 6
Figure 6
Cytokines produced by colon tissue in DSS-induced colitis model. Mice were orally administered with LL-pILMAM, LL-pILEMPTY or PBS 7 days before and during colitis induction. Colitis was induced by adding DSS in drinking water during 7 days (D0-D7). Then DSS was removed from drinking water and mice allowed to recover during 5 days (D7-D12). At D12 mice were killed, colon was withdrawn and proteins extracted. Cytokine concentration in protein extracts was monitored by ELISA. **P < 0.01, ***P < 0.001.
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References

    1. Almeida J. F., Mariat D., Azevedo V., Miyoshi A., de Moreno de LeBlanc A., Del Carmen S., et al. . (2014). Correlation between fibronectin binding protein A expression level at the surface of recombinant Lactococcus lactis and plasmid transfer in vitro and in vivo. BMC Microbiol. 14:248. 10.1186/s12866-014-0248-9 - DOI - PMC - PubMed
    1. Atarashi K., Tanoue T., Oshima K., Suda W., Nagano Y., Nishikawa H., et al. . (2013). Treg induction by a rationally selected mixture of Clostridia strains from the human microbiota. Nature 500, 232–236. 10.1038/nature12331 - DOI - PubMed
    1. Aubry C., Michon C., Chain F., Chvatchenko Y., Goffin L., Zimmerli S. C., et al. . (2015). Protective effect of TSLP delivered at the gut mucosa level by recombinant lactic acid bacteria in DSS-induced colitis mouse model. Microb. Cell Fact. 14, 176. 10.1186/s12934-015-0367-5 - "V体育安卓版" DOI - PMC - PubMed
    1. Chassaing B., Aitken J. D., Malleshappa M., Vijay-Kumar M. (2014). Dextran sulfate sodium (DSS)-induced colitis in mice. Curr. Protoc. Immunol. 104:15.215. 10.1002/0471142735.im1525s104 - DOI - PMC - PubMed
    1. Chatel J. M., Pothelune L., Ah-Leung S., Corthier G., Wal J. M., Langella P. (2008). In vivo transfer of plasmid from food-grade transiting lactococci to murine epithelial cells. Gene Ther. 15, 1184–1190. 10.1038/gt.2008.59 - "VSports" DOI - PubMed