Toll-like receptor-2-activating bifidobacteria strains differentially regulate inflammatory cytokines in the porcine intestinal epithelial cell culture system: finding new anti-inflammatory immunobiotics

Abstract

A total of 23 strains of bifidobacteria taxonomically belonging to five species were tested for their potent immunomodulatory effect using a combination of two methods: the NF-κB-reporter assay using a toll-like receptor 2-expressing transfectant (HEK(pTLR2) system) and the mitogenic assay using porcine Peyer's patches immunocompetent cells. Among the four preselected strains from different immunomodulatory groups, Bifidobacterium breve MCC-117 was able to efficiently modulate the inflammatory response triggered by enterotoxigenic Escherichia coli (ETEC) in a porcine intestinal epithelial (PIE) cell line. Moreover, using PIE cells and swine Peyer's patches immunocompetent cell co-culture system, we demonstrated that the immunoregulatory effect of B. breve MCC-117 was related to the capacity of the strain to influence PIE and immune cell interactions, leading to the stimulation of regulatory T cells. The results suggested that bifidobacteria that express high activity in both the HEK(pTLR2) and the mitogenic assays may behave like potential anti-inflammatory strains. The combination of the HEK(pTLR2) system, the evaluation of mitogenic activity and PIE cells will be of value for the development of new immunologically functional foods and feeds that could prevent inflammatory intestinal disorders. Although our findings should be proven in appropriate experiments in vivo, the results of the present work provide a scientific rationale for the use of B. breve MCC-117 to prevent ETEC-induced intestinal inflammation.

Immunobiotic Lactobacillus jensenii Elicits Anti-Inflammatory Activity in Porcine Intestinal Epithelial Cells by Modulating Negative Regulators of the Toll-Like Receptor Signaling Pathway

“…A recent study in Caco-2 cells demonstrated that L. rhamnosus GG is able to counteract ETEC-induced upregulation of IL-1␤ and TNF-␣ and the downregulation of TGF-␤1 expression, and consequently it prevents cytokine deregulation (31). In addition, comparative studies between L. rhamnosus GG and Bifidobacterium animalis MB5 (31) and between different bifidobacterial strains (8) demonstrated that the effects of individual probiotic strains on the inflammatory responses in IECs differ (8,31). These findings indicate that some probiotic LAB strains may be beneficial for preventing inflammatory response-mediated damage during weaning.…”

The toll-like receptor family protein RP105/MD1 complex is involved in the immunoregulatory effect of exopolysaccharides from Lactobacillus plantarum N14

“…Next, harvested PP or MLN cells were subjected to hypertonic rescue in an equal volume of 1.5% NaCl. Finally, immune cells were fractionated using Lympholyte-mammal (Cedarlane, Hornby, Ontario, Canada) density gradient centrifugation, and the isolated immune cells were suspended in complete RPMI 1640 (Sigma, St. Louis, MO, USA) supplemented with 10% FCS (Sigma), 50 g/mL and 50 g/mL penicillin-streptomycin (Gibco ® , Life Tech., Carlsbad, CA, USA) (Fujie et al, 2011).…”

“…The mitogenicity assay was performed as described previously (Fujie et al, 2011). Briefly, PPs or MLN immunocompetent cells (2 × 10 5 cells/well) were placed in a round ninety-six-well microplate (SUMILON ® , Sumitomo Bakelite Co. Ltd., Tokyo, Japan) and incubated at 5% CO 2 , 37 • C for 48 h in complete RPMI 1640 medium (Sigma, St. Louis, MO, USA) supplemented with 2% FCS.…”

Immunobiotic Lactobacillus jensenii Modulates the Toll-Like Receptor 4-Induced Inflammatory Response via Negative Regulation in Porcine Antigen-Presenting Cells

“…Two-step real-time quantitative PCR (qPCR) was used to characterize the expression of specific mRNAs in immune cells (13,28). Total RNA was isolated from individual samples of porcine APCs using TRIzol reagent.…”