Saccharomyces boulardii modulates dendritic cell properties and intestinal microbiota disruption after antibiotic treatment

Abstract

Saccharomyces boulardii is a non-pathogenic yeast with biotherapeutic properties that has been used successfully to prevent and to treat various infectious and antibiotic-associated diarrheas. The intestinal microbiota is responsible for colonization resistance and immune response to pathogens but can be disrupted by antibiotics and lose its barrier effect. Dendritic cells (DCs) are professional antigen-presenting cells of the immune system with the ability to initiate a primary immune response or immune tolerance. In a human microbiota-associated mouse model, we evaluated the influence of S. boulardii on the composition of the microbiota and on the properties of dendritic cells in normal homeostatic conditions and after antibiotic-induced stress. The DCs were derived from splenic precursors. Membrane antigen expression and phagocytosis of FITC-latex beads by DCs were evaluated by flow cytometry. The molecular analysis of the microbiota was performed with fluorescence in situ hybridization (FISH) combined with flow cytometry or confocal microscopy using group specific 16S rRNA targeted probes. This evaluation was conducted during and after a 7-day oral treatment with amoxicillin-clavulanic acid alone and in combination with the administration of the yeast. The antibiotic treatment increased the phagocytic activity of DCs. Their antigen presenting function (MHC class II antigen and CD 86 costimulatory molecule membrane expression) was up-regulated. This reflects a functional activation of DCs. In the presence of S. boulardii, the modification of membrane antigen expression was down regulated. To correlate these modifications to the microbiota disruption, we analyzed in parallel the composition of the intestinal microbiota. As previously shown, the amoxicillin-clavulanic acid treatment, both alone and with S. boulardii, did not quantitatively alter the total microbiota. In contrast, after one day of the antibiotic treatment the Clostridium coccoides group decreased dramatically in the two groups of mice treated with the antibiotic. The level then increased regularly, and at days 17, 22 and 24 it increased faster (P < 0.05) in the AB+ Sb group than in the AB group, reaching the initial level at day 29. The Bacteroides group in the two groups of mice increased during the antibiotic treatment and decreased after the antibiotic was stopped, reaching the initial level. The rate of decrease was faster for the AB+ Sb group than for the AB group, with a significant difference (P < 0.05) at days 17 and 22. During antibiotic treatment, the Enterobacteriaceae group became detectable and its level increased in both groups of mice. After discontinuation of the antibiotic, its level decreased to become undetectable at day 29, without significant difference between the two groups. These results showed that S. boulardii treatment tends to restore the balance of the dominant anaerobic microbiota more rapidly in human microbiota associated-mice treated with amoxicillin-clavulanic acid; the results also suggest that the yeast has a role in modulating the specific immune response to microbial associated-molecular patterns. This may explain, at least in part, the beneficial effects of S. boulardii in preventing antibiotic-associated diarrhea. This also suggests that the yeast plays a role in maintaining intestinal homeostasis.