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. 2018 Dec 1;125(6):1881-1889.
doi: 10.1111/jam.14069. Epub 2018 Aug 14.

Bifidobacterium animalis ssp. lactis Bb12 induces IL-10 through cell membrane-associated components via TLR2 in swine

M Arenas-Padilla  1 , J L Duarte-Gutiérrez  1 , V Mata-Haro  1
Affiliations

Bifidobacterium animalis ssp. lactis Bb12 induces IL-10 through cell membrane-associated components via TLR2 in swine

M Arenas-Padilla et al. J Appl Microbiol. .

Abstract

Aim: To investigate the role of Toll-like receptor 2 (TLR2) in interleukin-10 (IL-10) production induced by Bifidobacterium animalis ssp. lactis Bb12 (Bb12) in swine immune cells.

Methods and results: Blood-monocytes and cells from mesenteric lymph nodes were obtained from pigs and cultured with live Bb12 for 4 and 12 h. Transcript levels of IL-10 and TLR2 were analysed. Furthermore, TLR2 was blocked to determine its participation in IL-10 production. TLR2 blockade was achieved with neutralizing antibodies, followed by stimulation with Bb12. Bifidobacteria induced IL-10 production in both swine monocytes and mesenteric cells. Monocytes with TLR2 blockade had a decrease in IL-10 transcripts, while mesenteric cells did not. Bacterial cell wall components were responsible for Bb12-induced IL-10 production since no IL-10 was detected in the culture supernatant.

Conclusions: We demonstrated that IL-10 production is largely mediated through the recognition of Bb12 structures by TLR2, as bacterial metabolites in the culture supernatant failed to induce IL-10 expression.

Significance and impact of the study: The present study provides evidence for the potential use of Bb12 in the swine industry; these bacteria can also be used as additional method to treat intestinal inflammation and enhance intestinal health in pigs.

Keywords: Bifidobacterium animalis ssp. lactis Bb12; IL-10; TLR2; monocytes; probiotics; swine.

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Conflict of interest statement

The authors declare that there is no conflict of interest.

Figures

Figure 1
Figure 1
Bb12 induces IL‐10 expression in swine cells. Monocytes and mesenteric lymph node (MLN) cells were cultured in presence of Bb12 for 4 h, and the transcript levels of IL‐10 (a), or for 12 h and IL‐10 production were measured in culture supernatants (b). Swine monocytes and MLN cells were cultured in the presence of Bb12 for 4 h, and the relative expression of TLR2 was measured (c). RT‐qPCR results were normalized using expression level of swine PPIA gen. The data are shown as the means ± SD of three independent experiments (*P < 0·05).
Figure 2
Figure 2
Bb12 induced IL‐10 is mediated by TLR2. Monocytes were treated with an anti‐TLR2 neutralizing antibody (40 μg ml−1) prior to Bb12 stimulation for 2 and 4 h, and the transcript level of IL‐10 (a) and NF‐κB (b) was measured by RT‐qPCR. The results were normalized using expression level of swine PPIA gen. The data are shown as the means ± SD of three independent experiments (***P < 0·001).
Figure 3
Figure 3
IL‐10 production is mediated through the interaction of bacterial cell wall components with swine monocytes. Monocytes were stimulated with Bb12 culture supernatant for 4 h, and the transcript levels of IL‐10 (a) and TLR2 (b) were measured. RT‐qPCR results were normalized using expression level of swine PPIA gen. The data are shown as the means ± SD of three independent experiments (*P < 0·05).
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