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. 2015:2015:361604.
doi: 10.1155/2015/361604. Epub 2015 Feb 24.

Impact of kefir derived Lactobacillus kefiri on the mucosal immune response and gut microbiota

P Carasi  1 , S M Racedo  2 , C Jacquot  2 , D E Romanin  3 , M A Serradell  4 , M C Urdaci  2
Affiliations

Impact of kefir derived Lactobacillus kefiri on the mucosal immune response and gut microbiota

P Carasi et al. J Immunol Res. 2015.

Abstract

The evaluation of the impact of probiotics on host health could help to understand how they can be used in the prevention of diseases. On the basis of our previous studies and in vitro assays on PBMC and Caco-2 ccl20:luc reporter system presented in this work, the strain Lactobacillus kefiri CIDCA 8348 was selected and administrated to healthy Swiss mice daily for 21 days. The probiotic treatment increased IgA in feces and reduced expression of proinflammatory mediators in Peyer Patches and mesenteric lymph nodes, where it also increased IL-10. In ileum IL-10, CXCL-1 and mucin 6 genes were upregulated; meanwhile in colon mucin 4 was induced whereas IFN-γ, GM-CSF, and IL-1β genes were downregulated. Moreover, ileum and colon explants showed the anti-inflammatory effect of L. kefiri since the LPS-induced increment of IL-6 and GM-CSF levels in control mice was significantly attenuated in L. kefiri treated mice. Regarding fecal microbiota, DGGE profiles allowed differentiation of experimental groups in two separated clusters. Quantitative PCR analysis of different bacterial groups revealed only significant changes in Lactobacillus population. In conclusion, L. kefiri is a good candidate to be used in gut inflammatory disorders.

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Figures

Figure 1
Figure 1
Modulation of proinflammatory response in Caco-2 ccl20:luc reporter system by L. kefiri strains. NAL: normalized average luminescence expressed as percentage of activity induced with flagellin stimulation; FliC: Salmonella-isolated flagellin; Basal: without any stimulation. Results are expressed as mean ± standard deviation and are representative of at least three independent experiments. * P < 0.01.
Figure 2
Figure 2
IgA quantification from fecal samples taken on day 7, 14, or 21 from control mice and L. kefiri treated mice (Lk). Results are expressed as mean ± standard deviation. * P < 0.05.
Figure 3
Figure 3
Gene expression ratio in ileum (black) and colon (white) of Lk group versus control group after 7 days of L. kefiri administration. The x-axis of the plot represents log2 relative expression level of the gene and the y-axis displays the −log10 P (statistical significance). The names of the genes which displayed significant differences are included.
Figure 4
Figure 4
Gene expression ratio of Lk group versus control group after 21 days of L. kefiri administration. The x-axis of the plot represents log2 relative expression level of the gene and the x-axis displays the −log10 P (statistical significance). The names of the genes which displayed significant differences are included. (a) Expression in ileum (black) and colon (white). (b) Expression in PP (black) and mLN (white).
Figure 5
Figure 5
Cytokine's release in supernatants of (a) ileum and (b) colonic explants cultured for 24 h in the presence of LPS. Results are expressed as mean ± standard deviation. * P < 0.05.
Figure 6
Figure 6
Evaluation of microbiota on fecal samples taken on the 21st day of trial from control and Lk groups. (a) Total bacteria DGGE profiles of five mice from control group (lanes C1 to C5) and five from Lk group (lanes L1 to L5). (b) Dendrogram for the total bacterial DGGE profiles. Clustering analysis was performed using the UPGMA linkage. (c) qPCR quantification of total bacteria, Firmicutes, Bacteroidetes, and Lactobacillus spp. Results are expressed as mean ± standard deviation. * P < 0.05.
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