A Lactobacillus rhamnosus GG-derived Soluble Protein, p40, Stimulates Ligand Release from Intestinal Epithelial Cells to Transactivate Epidermal Growth Factor Receptor

Abstract

Background: p40 is a Lactobacillus rhamnosus GG-derived protein.Results: p40 stimulates ADAM17 activation and HB-EGF release, which is required for EGF receptor transactivation, prevention of apoptosis, and preservation of barrier function in intestinal epithelial cells. Conclusion: p40 transactivates the EGF receptor through ADAM17-mediated HB-EGF release in intestinal epithelial cells. Significance: These results define a mechanism of p40 in modulating intestinal epithelial cell homeostasis.

Activation of Epidermal Growth Factor Receptor Mediates Mucin Production Stimulated by p40, a Lactobacillus rhamnosus GG-derived Protein

“…We have reported that EGFR transactivation by p40 occurs through the release of an EGFR ligand, HB-EGF. Thus, p40 stimulates a delayed (30 min) and longer period of EGFR activation (up to 2 h), compared with HB-EGF (15-30 min) (24). This study further showed that p40 stimulated several peaks of EGFR activation, which are correlated with increased Muc2 gene expression at these time points in LS174T cells.…”

“…To elucidate the mechanisms of probiotic action, we purified and cloned p40 from LGG culture supernatants. p40 has beneficial effects on colon epithelial homeostasis through activation of EGFR and Akt in intestinal epithelial cells (23)(24)(25). In the present study, we demonstrate that p40 promotes Muc2 gene expression and MUC2 production in LS174T cells, which requires EGFR activation.…”

“…p40 Activates EGFR and Up-regulates Mucin Production in LS174T Cells-Our previous studies have demonstrated that p40 transactivates EGFR by stimulating HB-EGF release in intestinal epithelial cells (24), which is required for prevention and treatment of experimental colitis in mice (25). However, whether p40 transactivates EGFR in goblet cells and stimulates mucin production as a mucosal defense mechanism against injury and inflammatory assault in then intestinal tract is unknown.…”

“…We have previously purified and cloned a LGG-derived soluble protein, p40, that prevents cytokine-induced epithelial damage and apoptosis, as well as hydrogen peroxide disruption of the epithelial barrier function through stimulation of EGFR ligand release, which activates EGFR in intestinal epithelial cells (22)(23)(24). Furthermore, specific delivery of p40 to the colon using hydrogel beads to protect it from degradation prevents and treats colonic epithelial cell injury and inflammation in mouse models of colitis in an EGFR-dependent manner (25).…”

Sirtuin-6 Preserves R-spondin-1 Expression and increases Resistance of intestinal Epithelium to injury in Mice

“…Briefly, 96-well microtiter plates were (passage 7-15) were used according to the protocol described by Whitehead et al (17). Briefly, the cells were maintained in RPMI 1640 medium (Thermo Fisher Scientific) containing 5% heatinactivated fetal bovine serum (Thermo Fisher Scientific), 5 U/mL mouse interferon-γ (IFN-γ; PeproTech, Rocky Hill, NJ, USA), 100 unit/mL penicillin, 100 μg/mL streptomycin (Thermo Fisher Scientific), 5 μg/mL insulin, 5 μg/mL transferrin and 5 ng/mL selenous acid (#354351; BD Biosciences, San Jose, CA, USA) at 33°C as permissive conditions in a water-saturated atmosphere incubator with 5% CO 2 (18). Before experiments, the medium was replaced with one without IFN-γ and the cells were moved to 37°C (a nonpermissive temperature) for 24 h to allow them to differentiate.…”