The elastase and melanogenesis inhibitory and anti-inflammatory activities of phosvitin phosphopeptides produced using high-temperature and mild-pressure (HTMP) pretreatment and enzyme hydrolysis combinations

This study aimed to determine the skin protective effect of egg yolk phosvitin phosphopeptides (PPPs). Phosvitin was separated from the egg yolk, and PPPs were produced using high-temperature and mild-pressure (HTMP) pretreatment and enzyme-sterilization hydrolysis combinations. The elastase and melanogenesis inhibitory activities and anti-inflammatory effects of egg yolk PPPs were determined. All PPPs significantly inhibited elastase activity, but the PPPs prepared with HTMP pretreatment and trypsin-sterilization (HTMP-T-S) combination suppressed the tyrosinase activity the most. PPPs (3 mg/mL) inhibited the α-melanocyte-stimulating hormone-induced melanin production in B16F10 melanoma cells by 31.18 to 38.58%. In addition, PPPs effectively inhibited nitric oxide (NO) production in the LPS (lipopolysaccharide)-stimulated RAW 264.7 macrophages, and the PPPs from HTMP-T-S exhibited the highest inhibitory activity. The protein expressions of pro-inflammatory enzymes, inducible nitric oxide synthase, and cyclooxygenase-2 were down-regulated by the PPPs from the HTMP-T-S. Therefore, PPPs could be used as an anti-melanogenic, anti-elastase, and anti-inflammatory agent for humans and skin care products.This article is published as Lee, Ji-Eun, Bong Jeun An, Cheorun Jo, Byungrok Min, Hyun-Dong Paik, and Dong Uk Ahn. "The elastase and melanogenesis inhibitory and anti-inflammatory activities of phosvitin phosphopeptides produced using HTMP pre-treatment and enzyme hydrolysis combinations." Poultry Science 102 (2023): 102680. doi:10.1016/j.psj.2023.102680. Posted with permission.This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

The elastase and melanogenesis inhibitory and anti-inflammatory activities of phosvitin phosphopeptides produced using high-temperature and mild-pressure (HTMP) pretreatment and enzyme hydrolysis combinations

ABSTRACT: This study aimed to determine the skin protective effect of egg yolk phosvitin phosphopeptides (PPPs). Phosvitin was separated from the egg yolk, and PPPs were produced using high-temperature and mild-pressure (HTMP) pretreatment and enzyme-sterilization hydrolysis combinations. The elastase and melanogenesis inhibitory activities and anti-inflammatory effects of egg yolk PPPs were determined. All PPPs significantly inhibited elastase activity, but the PPPs prepared with HTMP pretreatment and trypsin-sterilization (HTMP-T-S) combination suppressed the tyrosinase activity the most. PPPs (3 mg/mL) inhibited the α-melanocyte-stimulating hormone-induced melanin production in B16F10 melanoma cells by 31.18 to 38.58%. In addition, PPPs effectively inhibited nitric oxide (NO) production in the LPS (lipopolysaccharide)-stimulated RAW 264.7 macrophages, and the PPPs from HTMP-T-S exhibited the highest inhibitory activity. The protein expressions of pro-inflammatory enzymes, inducible nitric oxide synthase, and cyclooxygenase-2 were down-regulated by the PPPs from the HTMP-T-S. Therefore, PPPs could be used as an anti-melanogenic, anti-elastase, and anti-inflammatory agent for humans and skin care products

Ulmus davidiana var. japonica Nakai upregulates eosinophils and suppresses Th1 and Th17 cells in the small intestine.

The bark of Ulmus davidiana var. japonica Nakai (Ulmaceae) has been used in traditional Korean medicine for chronic inflammation in the gastrointestinal tract. Here we investigated the frequency and cytokine profile of the major immune cells in the small intestinal lamina propria (SI LP), spleen, and mesenteric lymph nodes (MLNs) of mice treated orally with Ulmus davidiana var. japonica Nakai bark water extract (UDE) to address the immunomodulatory role of this herb in intestinal homeostasis. B6 mice were given 5g/kg UDE once daily for 14 days. They were then sacrificed, and cells were isolated from the spleen, MLNs, and SI LP. The proportion of B versus T lymphocytes, CD4(+) versus CD8(+) T lymphocytes, Th1 and Th17 cells, and Foxp3(+) regulatory T cells in the spleen, MLNs, and SI LP were analyzed. The frequency of antigen-presenting cells (APCs), including dendritic cells, macrophages, and eosinophils in the SI LP and the expression of costimulatory molecules on APCs were also evaluated. The numbers and frequencies of Th1 and Th17 cells in the SI LP were significantly reduced in the UDE-treated mice compared with PBS controls. In addition, the proportion of IL-4-producing eosinophils in the SI LP was significantly elevated in the UDE-treated mice compared with controls. Taken together, these data indicate that UDE up-regulates the number and frequency of SI LP eosinophils, which can down-regulate the Th1 and Th17 responses via IL-4 secretion and contribute to intestinal homeostasis

Extracellular vesicle-derived protein from Bifidobacterium longum alleviates food allergy through mast cell suppression

Background: The incidence of food allergies has increased dramatically during the last decade. Recently, probiotics have been studied for the prevention and treatment of allergic disease. Objective: We examined whether Bifidobacterium longum KACC 91563 and Enterococcus faecalis KACC 91532 have the capacity to suppress food allergies. Methods: B longum KACC 91563 and E faecalis KACC 91532 were administered to BALB/c wild-type mice, in which food allergy was induced by using ovalbumin and alum. Food allergy symptoms and various immune responses were assessed. Results: B longum KACC 91563, but not E faecalis KACC 91532, alleviated food allergy symptoms.Extracellular vesicles of B longum KACC 91563 bound specifically tomast cells and induced apoptosis without affecting T-cell immune responses. Furthermore, injection of family 5 extracellular solute-binding protein, a main component of extracellular vesicles, into micemarkedly reduced the occurrence of diarrhea in a mouse food allergy model. Conclusion: B longum KACC 91563 induces apoptosis of mast cells specifically and alleviates food allergy symptoms. Accordingly, B longum KACC 91563 and family 5 extracellular solute-binding protein exhibit potential as therapeutic approaches for food allergies. (c) 2015 American Academy of Allergy, Asthma & Immunology112171sciescopu

Structural specificities of cell surface β-glucan polysaccharides determine commensal yeast mediated immuno-modulatory activities

Yeast is an integral part of mammalian microbiome, and like commensal bacteria, has the potential of being harnessed to influence immunity in clinical settings. However, functional specificities of yeast-derived immunoregulatory molecules remain elusive. Here we find that while under steady state, β-1,3-glucan-containing polysaccharides potentiate pro-inflammatory properties, a relatively less abundant class of cell surface polysaccharides, dubbed mannan/β-1,6-glucan-containing polysaccharides (MGCP), is capable of exerting potent anti-inflammatory effects to the immune system. MGCP, in contrast to previously identified microbial cell surface polysaccharides, through a Dectin1-Cox2 signaling axis in dendritic cells, facilitates regulatory T (Treg) cell induction from naïve T cells. Furthermore, through a TLR2-dependent mechanism, it restrains Th1 differentiation of effector T cells by suppressing IFN-γ expression. As a result, administration of MGCP display robust suppressive capacity towards experimental inflammatory disease models of colitis and experimental autoimmune encephalomyelitis (EAE) in mice, thereby highlighting its potential therapeutic utility against clinically relevant autoimmune diseases.11Ysciescopu

Gut-specific Delivery of T-helper 17 Cells Reduces Obesity and Insulin Resistance in Mice

Background & Aims: Obesity and metabolic syndrome have been associated with alterations to the intestinal microbiota. However, few studies examined the effects of obesity on the intestinal immune system. We investigated changes in subsets of intestinal CD4+ Thelper (Th) cells with obesity and the effects of gut-tropic Th17 cells in mice on a high-fat diet (HFD). Methods: We isolated immune cells from small intestine and adipose tissue of C57BL/6 mice fed a normal chow diet or a HFD for 10 weeks and analyzed the cells by flow cytometry. Mice fed a vitamin A-deficient HFD were compared with mice fed a vitamin Asufficient HFD. Obese RAG1-deficient mice were given injections of only T regulatory cells or a combination of T regulatory cells and Th17 cells (wild type or deficient in integrin b7 subunit-deficient or interleukin 17 [IL17]). Mice were examined for weight gain, fat mass, fatty liver, glucose tolerance, and insulin resistance. Fecal samples were collected before and after T-cell transfer and analyzed for microbiota composition by metagenomic DNA sequencing and quantitative PCR. Results: Mice placed on a HFD became obese, which affected the distribution of small intestinal CD4+ TH cells. Intestinal tissues from obese mice had significant reductions in the proportion of Th17 cells but increased proportion of Th1 cells, compared with intestinal tissues from non-obese mice. Depletion of vitamin A in obese mice further reduced the proportion of Th17 cells in small intestine; this reduction correlated with more weight gain and worsening of glucose intolerance and insulin resistance. Adoptive transfer of in vitrodifferentiated gut-tropic Th17 cells to obese mice reduced these metabolic defects, which required the integrin b7 subunit and IL17. Delivery of Th17 cells to intestines of mice led to expansion of commensal microbes associated with leanness. Conclusions: In mice, intestinal Th17 cells contribute to development of a microbiota that maintains metabolic homeostasis, via IL17. Gut-homing Th17 cells might be used to reduce metabolic disorders in obese individuals.9

Significant increase of IL-4-expressing eosinophils in the SI LP of UDE-administered mice.

<div><p>(A) Cells isolated from SI LP were stained with CD11b, CD11c, and MHC class II and analyzed by flow cytometry. MHC class II ^lowCD11b^high CD11c^int cells (R1) are eosinophils. Among MHC class II^high cells, CD11b ^intCD11c^int cells (R4) and CD11c^high cells (R2 and R3) correspond to macrophages and dendritic cells, respectively.</p> <p>(B) Proportion and number of eosinophils, macrophages, and dendritic cells in the SP of UDE- versus PBS-administered control mice.</p> <p>(C) Antigen presenting cells isolated from SI LP were stained with CD40, CD80, and CD86 and analyzed by flow cytometry. Empty, solid line histograms represent cells from UDE-administered (red) versus PBS-administered control mice (black). Shaded histograms represent cells stained with isotype control of UDE-administered (dark gray) versus PBS-administered control mice (gray).</p> <p>(D) Mean fluorescence intensity of IL-4/GFP expressed by APCs from the SI LP of 4get mice were analyzed by flow cytometry.</p> <p>(E) Analysis of CD11b^highCD11c^int cells from LP of WT and ΔdblGATA mice for eosinophil markers CCR3 and Siglec F.</p> <p>*p<0.05, **p < 0.01, NS= not significant, LP: lamina propria, EO: eosionophil, DC: dendritic cell, Mϕ: macrophage, MFI: mean fluorescence intensity.</p></div

Decreased number of CD4⁺ T cells in the SI LP of UDE-administered mice.

<div><p>(A) Cells isolated from spleen (SP), mesenteric lymph node (MLN), and small intestinal lamina propria (SI LP) were stained with TCRβ and CD19 and analyzed by flow cytometry.</p> <p>(B) Frequency of T and B cells in the SP, MLN, and SI LP of UDE- versus PBS-administered control mice.</p> <p>(C) T Cells from SP, MLN, and SI LP were stained with CD4 and CD8α and analyzed by flow cytometry.</p> <p>(D) Frequency of CD4 and CD8 cells in the SP, MLN, and SI LP of UDE- versus PBS-administered control mice.</p> <p>(E) Total number of T/B cells and CD4/CD8 cells in the SP, MLN, and SI LP of UDE- versus PBS administered control mice.</p> <p>*p < 0.05, **p < 0.01, NS=not significant.</p></div

No significant differences of Th2 cells in the SI LP and serum total IgE, UDE-specific IgE and IgG1 levels between UDE- and PBS-administered mice.

<div><p>(A) Frequency and number of IL-4/GFP⁺ CD4⁺ T cells in the SI LP of UDE-versus PBS-administered control 4get mice.</p> <p>(B) Serum total IgE, UDE-specific IgE, and IgG1 secretion analyzed by ELISA.</p> <p>NS= not significant, OD₄₅₀=optical density at absorbance 450nm.</p></div

Cell surface polysaccharides of Bifidobacterium bifidum induce the generation of Foxp3(+) regulatory T cells

Dysregulation of intestinal microflora is linked to inflammatory disorders associated with compromised immunosuppressive functions of Foxp3(+) T regulatory (T-r(eg)) cells. Although mucosa-associated commensal microbiota has been implicated in T-reg generation, molecular identities of the "effector" components controlling this process remain largely unknown. Here, we have defined Bifidobacterium bifidum as a potent inducer of Foxp3(+) T-reg cells with diverse T cell receptor specificity to dietary antigens, commensal bacteria, and B. bifidum itself. Cell surface beta-glucan/galactan (CSGG) polysaccharides of B. bifidum were identified as key components responsible for T-reg induction. CSGG efficiently recapitulated the activity of whole bacteria and acted via regulatory dendritic cells through a partially Toll-like receptor 2-mediated mechanism. T-reg cells induced by B. bifidum or purified CSGG display stable and robust suppressive capacity toward experimental colitis. By identifying CSGG as a functional component of T-reg-inducing bacteria, our studies highlight the immunomodulatory potential of CSGG and CSGG-producing microbes © The Author