In addition, the ability of Lr1505 and Lr1506 to induce higher levels of MHCII and CD80/86 in poly(I:C)-challenged adherent cells was significantly blocked with anti-TLR2 antibodies (Figure 6B). Moreover, when studying the expression of IL-6, IFN-γ, IL-1β and IL-10 at post-translational levels in APCs stimulated with lactobacilli and then challenged with poly(I:C), MIF values remained at the same level of poly(I:C)-challenged control cells if the medium was added with anti-TLR2 antibodies (Figure 6B). In none selleck compound of the experiments performed here, anti-TLR9 antibodies exerted

any kind of effect on the expression of cytokines or molecules related to the antigen presenting process (Figure 6B). Figure 6 Role of toll-like receptor (TLR)-2 and TLR9 in

the immunoregulatory effect of immunobiotic lactobacilli in porcine intestinal epithelial (PIE) cells and antigen presenting cells (APCs) from Peyer’s patches in response to poly(I:C). Monocultures of PIE cells or adherent cells from Peyer’s patches were stimulated with Lactobacillus rhamnosus CRL1505 (Lr1505) or L. rhamnosus CRL1506 (Lr1506) with or without the addition of https://www.selleckchem.com/products/s63845.html anti-TLR2 or anti-TLR9 blocking antibodies. PIE and APCs were then challenged with poly(I:C). The mRNA expression of IFN-α, IFN-β, IL-6, MCP-1 and TNF-α in PIE and the mRNA expression of IFN-α, IFN-β, IL-1β, TNF-α, IFN-γ, IL-6, IL-2, IL-12, IL-10 and TGF-β in adherent cells was studied after 12 hours of poly(I:C) challenge (A). Cytokine mRNA levels were calibrated by the swine β-actin level and normalized by common logarithmic transformation. In addition, expression of MHC-II and CD80/86 molecules as well as intracellular levels of IL-1β, IL-10, IFN-γ and IL-10 (B) were studied in the three populations of APCs within adherent

cells defined with CD172a and CD11R1 markers. Values represent means and error bars indicate the standard deviations. The results out are means of 3 measures repeated 4 times with independent experiments. The mean differences among different superscripts letters were significant at the 5% level. Discussion Rotavirus represents one of the prevailing causes of infectious gastroenteritis in humans worldwide [3, 4, 6]. An initial and essential step in the viral infection cycle of rotavirus is entering and replicating in IECs of the small intestine [25]. IECs have been well defined as sentinels, because they are the first cells which encounter microorganisms and are not only a physical barrier but they recognize different types of PAMPs via PRRs, which are selectively expressed on the cell surface, internal compartments or cytoplasm. Upon virus internalization, dsRNA molecules are generated in infected cells [25]. These molecules are typical of many viral Doramapimod infections including rotavirus. Viral dsRNA activate PRRs such as TLR3, RIG-I, and MDA-5, which signal host cellular responses in order to try to control viral infection [25–27].