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Influence of environmental conditions and AI2 on the adhesion adaptive response of Lactobacillus acidophilus NCFM

ABSTRACT: This study reports an over 20-fold increase in the adhesive ability of Lactobacillus acidophilus NCFM to Caco-2 cells following a 1 hour incubation of cells that were concentrated ten-fold, immediately prior to adhesion. Microarray analysis of the global transcriptional response with and without exposure to the adhesion adaptive conditions revealed several genes potentially involved with adhesion to the intestinal epithelial cells and a classic stress response. Interestingly, putative genes linked to the synthesis of an interspecies signaling molecule, autoinducer-2 (AI-2), were overexpressed. Examination of the L. acidophilus NCFM genome revealed the complete pathway for AI-2 synthesis. AI-2 activity was detected in L. acidophilus NCFM during stationary growth phase using the Vibrio harveyi BB170 assay system. Using site-specific integration, an isogenic mutation was created in luxS and the resulting derivative of L. acidophilus NCFM did not produce AI-2. A 58 % decrease in adherence to Caco-2 cells was also observed by the LuxS- mutant when the cells were used for adhesion directly from logarithmic phase cultures. However, the LuxS- mutant strain still responded to adhesion adaptive conditions with significantly increased adherence indicating that additional factors contribute to the amplified adhesion response. Keywords: Culture response to specific environmental conditions

ORGANISM(S): Lactobacillus acidophilus

PROVIDER: GSE5674 | GEO | 2010/02/09

SECONDARY ACCESSION(S): PRJNA96955

REPOSITORIES: GEO

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Project description:Lactobacilli are probiotics that, among other health promoting effects, have been ascribed immunostimulating and virus preventive properties. Certain lactobacilli species have been shown to possess strong IL-12 inducing properties. As IL-12 production depends on the up-regulation of type I interferons, we hypothesized that the strong IL-12 inducing capacity of L. acidophilus NCFM in murine bone marrow derived DC is caused by an up-regulation of IFN-Î², which subsequently stimulates the induction of IL-12 and the dsRNA binding toll like receptor (TLR)-3. The expression of the genes encoding IFN-Î², IL-12, IL-10 and TLR-3 in DC upon stimulation with L. acidophilus NCFM was measured. L. acidophilus NCFM induced a much stronger expression of ifn-Î², il-12 and il-10 compared to the synthetic dsRNA ligand Poly I:C, whereas the levels of expressed tlr-3 were similar. By the use of whole genome microarray gene expression, we investigated whether other genes related to the viral defence were up-regulated in DC upon stimulation with L. acidophilus NCFM and found that various virus defence related genes, both early and late, were among the strongest up-regulated genes. The IFN-Î² stimulating capability was also detected in another L. acidophilus strain, but was not a property of other probiotic bacteria tested (B. bifidum and E. coli nissle).The IFN-Î² inducing capacity was markedly reduced in TLR-2 -/- DCs, dependent on endocytosis and the major cause of the induction of il-12 and tlr-3 in L. acidophilus NCFM stimulated cells. Collectively, our results reveal that certain lactobacilli trigger the expression of viral defence genes in DC in a TLR-2 manner through induction of IFN- Î². Experiment Overall Design: In the experiment Lactobacillus NCFM were added to murine dendritic cells and stimulated for 4, 10 or 18 hours. These were compared to control experiment at the same timepoints. Experiments were run in triplicates except for control 10h and control 18h which were only in duplicate, giving a total of 16 arrays.

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Influence of environmental conditions and AI2 on the adhesion adaptive response of Lactobacillus acidophilus NCFM

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