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

Abstract

Genome-wide transcriptional analysis in intestinal epithelial cells (IEC) can aid in elucidating the impact of single versus multi-strain probiotic combinations on immunological and cellular mechanisms of action. In this study we used human expression microarray chips in an in vitro intestinal epithelial cell model to investigate the impact of three probiotic bacteria, Lactobacillus helveticus R0052 (Lh-R0052), Bifidobacterium longum subsp. infantis R0033 (Bl-R0033) and Bifidobacterium bifidum R0071 (Bb-R0071) individually and in combination, and of a surface-layer protein (SLP) purified from Lh-R0052, on HT-29 cells’ transcriptional profile to poly(I:C)-induced inflammation. Hierarchical heat map clustering, Set Distiller and String analyses revealed that the effects of Lh-R0052 and Bb-R0071 diverged from those of Bl-R0033 and Lh-R0052-SLP. It was evident from the global analyses with respect to the immune, cellular and homeostasis related pathways that the co-challenge with probiotic combination (PC) vastly differed in its effect from the single strains and Lh-R0052-SLP treatments. The multi-strain PC resulted in a greater reduction of modulated genes, found through functional connections between immune and cellular pathways. Cytokine and chemokine analyses based on specific outcomes from the TNF-α and NF-κB signaling pathways revealed single, multi-strain and Lh-R0052-SLP specific attenuation of the majority of proteins measured (TNF-α, IL-8, CXCL1, CXCL2 and CXCL10), indicating potentially different mechanisms. These findings indicate a synergistic effect of the bacterial combinations relative to the single strain and Lh-R0052-SLP treatments in resolving toll-like receptor 3 (TLR3)-induced inflammation in IEC and maintaining cellular homeostasis, reinforcing the rationale for using multi-strain formulations as a probiotic.

A novel polyphenolic prebiotic and probiotic formulation have synergistic effects on the gut microbiota influencing Drosophila melanogaster physiology

“…The present study is among the first studies to directly compare the effectiveness of probiotics and a prebiotic in isolation to a formulation containing the probiotics or together as a synbiotic. As suggested by the several studies regarding probiotic and synbiotic formulations, the sum of its components is greater than their constituents [52]. We have confirmed this hypothesis as the synbiotic formulation was the most effective at inducing positive variations in the gut microbiota and these changes were reflected in the broad physiological traits of Drosophila including elevated fitness and motility in age.…”

Probiotics modulate gut microbiota and improve insulin sensitivity in DIO mice

“…The latter, whose definition is live microbial food supplements which help maintain the balance of intestinal microflora [18,19], have shown in some studies to be effective in the treatment of insulin resistance in both rodents and humans; however, the deep mechanisms that lead to such benefits are poorly understood [4,[18][19][20]. Indeed, multistrain probiotics appear to be efficient against a wide range of intestinal issues [21,22]. Despite the absence of strong evidence for metabolic disease, we hypothesized that probiotic mixture would be more suitable to test in an experimental model of obesity.…”

Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

“…Moreover, previous studies on L. helveticus R0052‐SLP showed in vitro immunomodulatory properties (MacPherson et al . ) as well as E. coli O157:H7 anti‐adhesion activity (Johnson‐Henry et al . ).…”

“…This is exemplified in the work of Pr evost and colleagues, who developed a bead-based assay for the detection of staphylococcal enterotoxin in food matrices using polyclonal antibodies following a specific patented preparation (Barasino et al 2012). Moreover, previous studies on L. helveticus R0052-SLP showed in vitro immunomodulatory properties (MacPherson et al 2017) as well as E. coli O157:H7 anti-adhesion activity (Johnson-Henry et al 2007). Therefore, identification of specific targets could be also useful for host-microbe interaction studies as surface-exposed protein might have a role in promoting health benefits.…”

“…Samples were washed once in DPBS by centrifugation 5 min at 9000 9 g to remove debris. Purified L. helveticus R0052 SLP was extracted using a 5 mol l À1 lithium chloride protocol adapted from previously established methods (Taverniti et al 2013;MacPherson et al 2017). Protein extracts were suspended in DPBS and stored at À20°C until use.…”