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

PLoS One. 2017 Jan 18;12(1):e0169847. doi: 10.1371/journal.pone.0169847. eCollection 2017.


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.

MeSH terms

  • Bifidobacterium longum subspecies infantis
  • Chemokines / genetics
  • Chemokines / metabolism
  • Cytokines / genetics
  • Cytokines / metabolism
  • Epithelial Cells
  • Gastroenteritis / drug therapy
  • Gastroenteritis / immunology
  • Gastroenteritis / metabolism*
  • Gene Expression Profiling
  • Gene Regulatory Networks / immunology*
  • Genome, Human
  • HT29 Cells
  • Humans
  • Inflammation Mediators / metabolism
  • Lactobacillus helveticus
  • Oligonucleotide Array Sequence Analysis
  • Probiotics / pharmacology*
  • Toll-Like Receptor 3 / immunology
  • Toll-Like Receptor 3 / metabolism*


  • Chemokines
  • Cytokines
  • Inflammation Mediators
  • TLR3 protein, human
  • Toll-Like Receptor 3

Grant support

This project was funded in its entirety by Lallemand Health Solutions Inc. The funding organization did not play a role in the study design, data collection and analysis, decision to publish or preparation of the manuscript and only provided financial support in the form of authors’ salaries and/or research materials. Moreover, all final research decisions were made by the Research Director of Lallemand Health Solutions Inc., Dr. Thomas Allan Tompkins, one of the author(s) of this manuscript. The funder Lallemand Health Solutions Inc. only provided financial support in the form of authors’ salaries and/or research materials.