Transcriptomic Analysis of the Innate Antiviral Immune Response in Porcine Intestinal Epithelial Cells: Influence of Immunobiotic Lactobacilli

Abstract

Lactobacillus rhamnosus CRL1505 and Lactobacillus plantarum CRL1506 are immunobiotic strains able to increase protection against viral intestinal infections as demonstrated in animal models and humans. To gain insight into the host–immunobiotic interaction, the transcriptomic response of porcine intestinal epithelial (PIE) cells to the challenge with viral molecular associated pattern poly(I:C) and the changes in the transcriptomic profile induced by the immunobiotics strains CRL1505 and CRL1506 were investigated in this work. By using microarray technology and reverse transcription PCR, we obtained a global overview of the immune genes involved in the innate antiviral immune response in PIE cells. Stimulation of PIE cells with poly(I:C) significantly increased the expression of IFN-α and IFN-β, several interferon-stimulated genes, cytokines, chemokines, adhesion molecules, and genes involved in prostaglandin biosynthesis. It was also determined that lactobacilli differently modulated immune gene expression in poly(I:C)-challenged PIE cells. Most notable changes were found in antiviral factors (IFN-α, IFN-β, NPLR3, OAS1, OASL, MX2, and RNASEL) and cytokines/chemokines (IL-1β, IL-6, CCL4, CCL5, and CXCL10) that were significantly increased in lactobacilli-treated PIE cells. Immunobiotics reduced the expression of IL-15 and RAE1 genes that mediate poly(I:C) inflammatory damage. In addition, lactobacilli treatments increased the expression PLA2G4A, PTGES, and PTGS2 that are involved in prostaglandin E2 biosynthesis. L. rhamnosus CRL1505 and L. plantarum CRL1506 showed quantitative and qualitative differences in their capacities to modulate the innate antiviral immune response in PIE cells, which would explain the higher capacity of the CRL1505 strain when compared to CRL1506 to protect against viral infection and inflammatory damage in vivo. These results provided valuable information for the deeper understanding of the host–immunobiotic interaction and their effect on antiviral immunity. The comprehensive transcriptomic analyses successfully identified a group of genes (IFN-β, RIG1, RNASEL, MX2, A20, IL27, CXCL5, CCL4, PTGES, and PTGER4), which can be used as prospective biomarkers for the screening of new antiviral immunobiotics in PIE cells and for the development of novel functional food and feeds, which may help to prevent viral infections.

Immunomodulatory Properties of Bacterium-Like Particles Obtained From Immunobiotic Lactobacilli: Prospects for Their Use as Mucosal Adjuvants

“…Transcriptomic studies revealed the capacity of L. rhamnosus CRL1505 to differentially modulate the innate antiviral immune response in porcine intestinal epithelial cells. Higher expression levels of type I IFNs as well as in several antiviral factors including ifit1, nlpr3, mda5, msx1, rig1 ifit2, and mx2 were found in CRL1505-treated cells when compared to control cells (23). In addition, it was demonstrated in vitro that viable L. rhamnosus CRL1505 is able to efficiently stimulate the expression of IFN-γ in porcine intestinal CD172a + CD11R1 − macrophages (22).…”

“…Our previous comparative in vitro studies evaluating the ability of L. rhamnosus CRL1505 and L. plantarum CRL1506 to modulate immune responses in intestinal epithelial cells and antigen presenting cells (APCs) found significant differences between the two lactobacilli strains (22,23). It was demonstrated that intestinal epithelial cells were modulated by immunobiotic CRL1505 and CRL1506 in a strain-dependent fashion to enhance antiviral responses (23). Interestingly, L. rhamnosus CRL1505 was more efficient that L. plantarum CRL1506 to increase the expression of IFN-β and IL-6 in intestinal epithelial cells, both cytokines known to influence immune responses generated by immune cells located under the epithelium.…”

An EGFP-marked recombinant lactobacillus oral tetravalent vaccine constitutively expressing α, ε, β1, and β2 toxoids for Clostridium perfringens elicits effective anti-toxins protective immunity

“…The delivery of vaccine antigens via viable vectors not only can stimulate the intestinal immune response, but can also produce specific immune responses against specific antigens that allow the body to achieve full protection. Compared to attenuated Salmonella as mucosal antigen delivery vehicles [36][37][38], lactic acid bacteria (LAB) are more advantageous [20,22,23,30,39]. The long-term use of L. casei in the food industry has been demonstrated to be non-pathogenic, and has been designated as "generally recognized as safe".…”

“…Lactobacillus is a Gram-positive, facultative anaerobic or microaerophilic, rod-shaped, non-spore-forming bacteria, which is the most common probiotic found in food and the intestinal tract of humans and animals [16]. Studies have shown that some specific isolates of lactobacilli have potential immunomodulatory properties, such as promoting strong adhesive interactions with intestinal epithelial cells [17] and preventing injury of the epithelial cell barrier [18], anti-inflammatory capacity [19], modulating innate immune response [20] and sIgA production [21], regulating immunological functions of dendritic cells, and T helper cells activation [22,23]. Moreover, lactobacilli can survive transit through the upper gastrointestinal tract and colonize the intestinal tract [24,25].…”

Lipoteichoic Acid Is Involved in the Ability of the Immunobiotic Strain Lactobacillus plantarum CRL1506 to Modulate the Intestinal Antiviral Innate Immunity Triggered by TLR3 Activation

“…Previous in vitro transcriptomic studies showed that Lactobacillus plantarum CRL1506 is able to modulate the expression of type I interferons (IFNs), antiviral factors, cytokines, chemokines, and adhesion molecules in porcine intestinal epithelial (PIE) cells challenged with the Toll-like receptor (TLR)-3 ligand poly(I:C) (9). In addition, studies in porcine antigen-presenting cells isolated from Peyer's patches demonstrated that the CRL1506 strain differentially modulated the expression of IFN-γ, interleukin (IL)-1β, IL-12, IL-6, and tumor necrosis factor (TNF)-α (10).…”