Multistrain Probiotic Modulation of Intestinal Epithelial Cells' Immune Response to a Double-Stranded RNA Ligand, Poly(I·C)

Abstract

A commercially available product containing three probiotic bacterial strains (Lactobacillus helveticus R0052, Bifidobacterium longum subsp. infantis R0033, and Bifidobacterium bifidum R0071) was previously shown in animal trials to modulate both T H 1 and T H 2 immune responses. Clinical studies on this combination of bacteria have also shown positive health effects against seasonal winter diseases and rotavirus infection. The goal of this study was to use a well-established in vitro intestinal epithelial (HT-29) cell model that has been shown to constitutively express double-stranded RNA (dsRNA) sensors (Toll-like receptor 3 [TLR3], retinoic acid-inducible gene I, melanoma differentiation-associated gene 5, and dsRNA-activated protein kinase). By using the HT-29 cell model, we wanted to evaluate whether or not this combination of three bacteria had the capacity to immune modulate the host cell response to a dsRNA ligand, poly(I·C). Using a custom-designed, two-color expression microarray targeting genes of the human immune system, we investigated the response of HT-29 cells challenged with poly(I·C) both in the presence and in the absence of the three probiotic bacteria. We observed that the combination of the three bacteria had a major impact on attenuating the expression of genes connected to proinflammatory T H 1 and antiviral innate immune responses compared to that obtained by the poly(I·C)-only challenge. Major pathways through which the multistrain combination may be eliciting its immune-modulatory effect include the TLR3 domain-containing adapter-inducing beta interferon (TRIF), mitogenactivated protein kinase, and NF-B signaling pathways. Such a model may be useful for selecting potential biomarkers for the design of future clinical trials.

Intestinal Innate Antiviral Immunity and Immunobiotics: Beneficial Effects against Rotavirus Infection

“…In this regard, we also reported that both B. infantis MCC12 and B. breve MCC1274 significantly reduced the expression of A20 in RVs-infected PIE cells (Figure 4) (50), which is in line with the capacity of both strains to improve IRF3 activation and IFN-β production. In line with our findings, MacPherson et al (64) also studied the effect of probiotics in the modulation of poly(I:C) induced inflammatory response in HT-29 cells. Stimulating HT29 cells with poly(I:C) alone increased the expression of A20, but the co-stimulation with poly(I:C) and probiotics significantly reduced A20 expression levels.…”

Genome-Wide Immune Modulation of TLR3-Mediated Inflammation in Intestinal Epithelial Cells Differs between Single and Multi-Strain Probiotic Combination

“…helveticus R0052 in a ratio of 20:20:60; respectively, and as described in MacPherson et al . [22]. To rehydrate the lyophilized bacteria for both single and multi-strain blend, 1 g was mixed for 15 min at room temperature (RT) in 99 mL of phosphate buffer [0.1% soy peptone (w/v), 0.121% K 2 HPO 4 (w/v), 0.034% KH 2 PO 4 (w/v)] as described in MacPherson et al .…”

“…and Audy et al . [22,27]. Briefly, bacterial pellet from 1 ml of this bacterial suspension was washed in PBS after centrifugation at 12800 x g for 10 min at room temperature (RT) and then re-suspended in serum-free RPMI-1640 media.…”

“…We have previously demonstrated through a custom-designed immune microarray analysis a multi-strain (Bl-R0033, Lh-R0052 and Bb-R0071) probiotic combination (PC) on TLR3-induced immune activation by polyinosinic:polycytidylic acid (poly(I:C)) in human intestinal epithelial cells attenuated T H 1 pro-inflammatory response through the TLR3-TRIF, MAPK and NF- K B signaling pathways [22]. However, recent evidence also indicates a single probiotic strain, L .…”

Modulation of the spleen transcriptome in domestic turkey (Meleagris gallopavo) in response to aflatoxin B1 and probiotics

“…Exposure to Lactobacillus in mice can increase 2′5′-OAS activity and can up-regulate the expression of multiple OAS family members (Solis-Pereyra et al 1997;Weiss et al 2010). However, the opposite effect was observed in human intestinal cells, where a mixture of Lactobacillus and Bifidobacteria prevented the up-regulation of OASL and other antimicrobial genes during viral (dsRNA) exposure (MacPherson et al 2014). A single highly significant transcript from TMC6, which likely encodes a transmembrane channel protein (Mutai et al 2005), was also down-regulated in all comparisons involving the PBAFB group, suggesting that the interaction of PB and AFB 1 has suppressive effects not seen in the PB group.…”