Epigenetic imprinting by commensal probiotics inhibits the IL-23/IL-17 axis in an in vitro model of the intestinal mucosal immune system

Lactic acid bacteria inhibits NFκB‐mediated transcriptional activation of IBD causing factors (IL‐23/IL‐17/CD40) by reducing histone acetylation while enhancing DNA methylation. The pathophysiology of IBD is characterized by a complex interaction between genes and the environment. Genetic and environmental differences are attributed to the heterogeneity of the disease pathway and to the epigenetic modifications that lead to altered gene expression in the diseased tissues. The epigenetic machinery consists of short interfering RNA, histone modifications, and DNA methylation. We evaluated the effects of Bifidobacterium breve (DSMZ 20213) and LGG (ATCC 53103), as representatives of commensal probiotics on the expression of IL‐17 and IL‐23, which play an important role in IBD, and on the epigenetic machinery in a 3D coculture model composed of human intestinal HT‐29/B6 or T84 cells and PBMCs. The cells were treated with LPS in the presence or absence of bacteria for 48 h, and the expression of IL‐17, IL‐23, and CD40 at the mRNA and protein levels was assessed using TaqMan qRT‐PCR and ELISA, respectively. Western blotting was used to assess the expression of the MyD88, the degradation of IRAK‐1 and IκBα, the expression of the NF‐κB p50/p65 subunits, the p‐p38 MAPK and p‐MEK1, as well as histone modifications. NF‐κB activity was assessed by NF‐κB‐dependent luciferase reporter gene assays. The accumulation of Ac‐H4 and DNA methylation was quantitatively assessed using colorimetric assays. B. breve and LGG diminished the LPS‐induced expression of IL‐17, IL‐23, CD40, and histone acetylation, while slightly enhancing DNA methylation. These effects were paralleled by a decrease in the nuclear translocation of NF‐κB, as demonstrated by a decrease in the expression of MyD88, degradation of IRAK‐1 and IκBα expression of the nuclear NF‐κB p50/p65 subunits, p‐p38 MAPK and p‐MEK1, and NF‐κB‐dependent luciferase reporter gene activity in LPS‐stimulated cells. B. breve and LGG may exert their anti‐inflammatory effects in the gut by down‐regulating the expression of the IBD‐causing factors (IL‐23/IL‐17/CD40) associated with epigenetic processes involving the inhibition of histone acetylation and the optimal enhancement of DNA methylation, reflected in the limited access of NF‐κB to gene promoters and reduced NF‐κB‐mediated transcriptional activation. We describe a new regulatory mechanism in which commensal probiotics inhibit the NF‐κB‐mediated transcriptional activation of I