As control, pups were treated with hydrogels without p40 (no-p40)


As control, pups were treated with hydrogels without p40 (no-p40). differentiation, and tight junction formation, and IgA production in early life in wild-type mice. These p40-induced effects were abolished in mice with specific deletion of EGFR in intestinal epithelial cells, suggesting that transactivation of EGFR in intestinal epithelial cells may mediate p40-regulated intestinal development. Furthermore, neonatal p40 treatment reduced the susceptibility to intestinal injury and colitis and promoted protective immune responses, including IgA production and differentiation of regulatory T cells, in adult mice. These findings reveal novel functions of neonatal supplementation of Fexinidazole probiotic-derived factors in promoting EGFR-mediated maturation of intestinal functions and innate immunity, which likely promote long-term beneficial outcomes. promotes juvenile growth during chronic undernutrition10. Furthermore, neonatal colonization of conventionally raised mice with GG (LGG) promotes growth, intestinal functional maturation, persistent IgA production, and decreased susceptibility to intestinal injury and colitis in adult mice11, suggesting that LGG colonization has an impact on intestinal development and long-term health outcomes in adulthood. Multiple mechanisms of probiotic action have been suggested, however, little Fexinidazole is known about precise mode of probiotic action12, 13. Several factors derived from the gut microbiota, including probiotic bacteria, have been identified as functional effectors. p40 was originally isolated from LGG culture supernatant by our group14. Production of this secretory protein is usually strain-specific14. Functional analysis of p40 revealed that p40 preserves barrier function, suppresses cytokine-induced apoptosis, promotes mucin production in intestinal epithelial cells, and enhances IgA production through up-regulation of the expression of a proliferation-inducing ligand (APRIL) in intestinal epithelial cells14C17, thereby, protecting the intestine from injury and inflammation in mice17. Further studies exhibited that transactivation of the epidermal growth factor receptor (EGFR) in intestinal epithelial cells through activation of a disintegrin and metalloproteinase domain-containing protein-17 (ADAM-17) for HB-EGF release serves as a signaling mechanism for regulating these protective cellular responses by p4018. As a member of the ErbB family of type 1 receptor tyrosine kinases, activation of EGFR stimulates a variety of downstream signaling pathways, such as Akt and mitogen-activated protein kinase, leading to several cellular responses, including cell proliferation, differentiation, migration, and survival19, 20. EGFR signaling is required for postnatal growth because insufficient EGFR signaling causes Fexinidazole death in the perinatal period, and surviving mice die from hemorrhagic enteritis in the postnatal period21. Studies have also shown that ErbB ligands, EGF22, 23 and HB-EGF24, are present in amniotic fluid and breast milk and are involved in stimulating intestinal epithelial cell growth. Growth and epithelial cell proliferation are enhanced by EGF-expressing in mice at weaning25. While some EGFR mutations are considered to be tumor promoters, EGF has shown therapeutic potential in human ulcerative colitis26 and EGFR activation ameliorates chronic inflammation, thus limiting colitis-associated tumorigenesis27. Here we show that neonatal p40 supplementation promotes functional maturation of the intestine and stimulates persistent IgA production from early life to adulthood, in an EGFR-dependent manner. Adult mice with p40 supplementation in early life exhibit decreased susceptibility to intestinal injury and inflammation. Thus, these results broaden Fexinidazole our understanding of the mechanisms underlying the effects of probiotics on intestinal development and immune responses in early life and long-term health outcomes. Results Development of a hydrogel system for delivering p40 to the small intestine and the colon in neonatal mice p40 used in this Fexinidazole study was purified from LGG culture supernatant, as previously reported14. The level of endotoxin in p40 isolates was 0.03 EU/g p40 protein. For protecting p40 from the harsh conditions in the gastrointestinal tract, such as the presence of proteases and an acidic environment in the stomach, Rabbit Polyclonal to TF2H1 a pectin and zein delivery system was generated in our previous study for specifically delivering p40 to the colon17. This colon-specific delivery system requires the high concentration of pectin, such as 6%.