Expression Of Host Defense Peptides Research Articles

CXCR2, as a key regulatory gene of HDP-PG-1, maintains intestinal mucosal homeostasis

The intestine defends against pathogenic microbial invasion via the secretion of host defense peptides (HDPs). Nutritional immunomodulation can stimulate the expression of endogenous HDPs and enhance the body's immune defense, representing a novel non-antibiotic strategy for disease prevention. The project aims to explore the regulatory mechanism of protegrin-1 (PG-1) expression using sodium phenylbutyrate (PBA) by omics sequencing technology and further investigate the role of key regulatory genes on intestinal health. The results showed that PBA promoted PG-1 expression in intestinal epithelial cells based on cell density through epidermal growth factor receptor (EGFR) and G protein-coupled receptor (GPR43). Transcriptome sequencing and microRNA sequencing revealed that C-X-C motif chemokine receptor 2 (CXCR2) exhibited interactions with PG-1. Pre-treatment cells with a CXCR2 inhibitor (SB225002) effectively suppressed the induction of PG-1 by PBA. Furthermore, SB225002 significantly suppressed the gene expression of HDPs in the jejunum of mice without influencing on the morphology, number of goblet cells, and proliferation of the intestine. CXCR2 inhibition significantly reduced the expression of HDPs during E. coli infection, and resulted in the edema of jejunal epithelial cells. The 16S rDNA analysis of cecal contents showed that the E. coli and SB225002 treatments changed gut microbiota diversity and composition at different taxonomic levels. Correlation analysis suggested a potential regulatory relationship between gut microbiota and HDPs. To that end, a gene involved in the HDP expression, CXCR2, has been identified in the study, which contributes to improving intestinal immune function. PBA may be used as a functional additive to regulate intestinal mucosal function, thereby enhancing the health of the intestinal and host.

Two intestinal microbiota-derived metabolites, deoxycholic acid and butyrate, synergize to enhance host defense peptide synthesis and alleviate necrotic enteritis

BackgroundNecrotic enteritis (NE) is a major enteric disease in poultry, yet effective mitigation strategies remain elusive. Deoxycholic acid (DCA) and butyrate, two major metabolites derived from the intestinal microbiota, have independently been shown to induce host defense peptide (HDP) synthesis. However, the potential synergy between these two compounds remains unexplored.MethodsTo investigate the possible synergistic effect between DCA and butyrate in regulating HDP synthesis and barrier function, we treated chicken HD11 macrophage cells and jejunal explants with DCA and sodium butyrate (NaB), either individually or in combination, for 24 h. Subsequently, we performed RNA isolation and reverse transcription-quantitative PCR to analyze HDP genes as well as the major genes associated with barrier function. To further determine the synergy between DCA and NaB in enhancing NE resistance, we conducted two independent trials with Cobb broiler chicks. In each trial, the diet was supplemented with DCA or NaB on the day-of-hatch, followed by NE induction through sequential challenges with Eimeria maxima and Clostridium perfringens on d 10 and 14, respectively. We recorded animal mortality after infection and assessed intestinal lesions on d 17. The impact of DCA and NaB on the microbiota in the ileum and cecum was evaluated through bacterial 16S rRNA gene sequencing.ResultsWe found that the combination of DCA and NaB synergistically induced multiple HDP genes in both chicken HD11 cells and jejunal explants. Additionally, the gene for claudin-1, a major tight junction protein, also exhibited synergistic induction in response to DCA and NaB. Furthermore, dietary supplementation with a combination of 0.75 g/kg DCA and 1 g/kg NaB led to a significant improvement in animal survival and a reduction in intestinal lesions compared to either compound alone in a chicken model of NE. Notably, the cecal microbiota of NE-infected chickens showed a marked decrease in SCFA-producing bacteria such as Bacteroides, Faecalibacterium, and Cuneatibacter, with lactobacilli becoming the most dominant species. However, supplementation with DCA and NaB largely restored the intestinal microbiota to healthy levels.ConclusionsDCA synergizes with NaB to induce HDP and claudin-1 expression and enhance NE resistance, with potential for further development as cost-effective antibiotic alternatives.

A frog-derived antimicrobial peptide as a potential anti-biofilm agent in combating Staphylococcus aureus skin infection.

Staphylococcus aureus (S. aureus), one of the most prevalent bacteria found in atopic dermatitis lesions, can induce ongoing infections and inflammation by downregulating the expression of host defence peptides in the skin. In addition, the emergence of the 'superbug' Methicillin-resistant S. aureus (MRSA) has made the treatment of these infections more challenging. Antimicrobial peptides (AMPs), due to their potent antimicrobial activity, limited evidence of resistance development, and potential immunomodulatory effects, have gained increasing attention as potential therapeutic agents for atopic dermatitis. In this study, we report a novel AMP, brevinin-1E-OG9, isolated from the skin secretions of Odorrana grahami, which shows potent antibacterial activity, especially against S. aureus. Based on the characteristics of the 'Rana Box', we designed a set of brevinin-1E-OG9 analogues to explore its structure-activity relationship. Brevinin-1E-OG9c-De-NH2 exhibited the most potent antimicrobial efficacy in both in vitro and ex vivo studies and attenuated inflammatory responses induced by lipoteichoic acid and heat-killed microbes. As a result, brevinin-1E-OG9c-De-NH2 might represent a promising candidate for the treatment of S. aureus skin infections.

Host Defense Peptides in Nutrition and Diseases: A Contributor of Immunology Modulation.

Host defense peptides (HDPs) are primary components of the innate immune system with diverse biological functions, such as antibacterial ability and immunomodulatory function. HDPs are produced and released by immune and epithelial cells against microbial invasion, which are widely distributed in humans, animals, plants, and microbes. Notably, there are great differences in endogenous HDP distribution and expression in humans and animals. Moreover, HDP expression could be regulated by exogenous substances, such as nutrients, and different physiological statuses in health and disease. In this review, we systematically assessed the regulation of expression and mechanism of endogenous HDPs from nutrition and disease perspectives, providing a basis to identify the specificity and regularity of HDP expression. Furthermore, the regulation mechanism of HDP expression was summarized systematically, and the differences in the regulation between nutrients and diseases were explored. From this review, we provide novel ideas targeted the immune regulation of HDPs for protecting host health in nutrition and practical and effective new ideas using the immune regulation theory for further research on protecting host health from pathogenic infection and excessive immunity diseases under the global challenge of the antibiotic-abuse-induced series of problems, including food security and microbial resistance.

Preventing Respiratory Viral Diseases with Antimicrobial Peptide Master Regulators in the Lung Airway Habitat

The vast surface area of the respiratory system acts as an initial site of contact for microbes and foreign particles. The whole respiratory epithelium is covered with a thin layer of the airway and alveolar secretions. Respiratory secretions contain host defense peptides (HDPs), such as defensins and cathelicidins, which are the best-studied antimicrobial components expressed in the respiratory tract. HDPs have an important role in the human body's initial line of defense against pathogenic microbes. Epithelial and immunological cells produce HDPs in the surface fluids of the lungs, which act as endogenous antibiotics in the respiratory tract. The production and action of these antimicrobial peptides (AMPs) are critical in the host's defense against respiratory infections. In this study, we have described all the HDPs secreted in the respiratory tract as well as how their expression is regulated during respiratory disorders. We focused on the transcriptional expression and regulation mechanisms of respiratory tract HDPs. Understanding how HDPs are controlled throughout infections might provide an alternative to relying on the host's innate immunity to combat respiratory viral infections.

Campylobacter jejuni infection induces dynamic expression of avian host defense peptides in vitro and in vivo

Campylobacter jejuni is considered as the leading cause of worldwide foodborne bacterial gastroenteritis. Chicken is the main reservoir of C. jejuni. Avian innate immune responses to C. jejuni remain poorly defined. Chicken host defense peptides (HDPs) are the major components of avian innate immune system. This study aimed to characterize the chicken HDPs responses to C. jejuni in vitro and in vivo. In the HD11 macrophage cell line, the HDPs, including AvBD1–2, CATH1–3, AvBD7, AvBD4, and AvBD6, were relatively higher expressed in untreated cells, whereas the expressions were suppressed after C. jejuni infection. In contrast, C. jejuni infection significantly increased the expression of the lower expressed HDPs, such as AvBD3, AvBD5, AvBD8–14, and CATHB1, in untreated cells. In the chicken challenge experiment, the immune tissues of spleens and cecal tonsils were collected from C. jejuni-infected and uninfected chickens at 1, 3 and 15 day post inoculation (DPI). In spleens of C. jejuni-infected chickens, only AvBD14 expression was elevated at 1 DPI. The majority of avian HDPs were significantly up-regulated at 3 DPI and dramatically decreased to the levels of uninfected controls at 15 DPI. In chicken cecal tonsils, only AvBD9 and AvBD14 were significantly up-regulated at 1 DPI with C. jejuni infection. Collectively, C. jejuni infection induced dynamic expression of chicken HDPs in both macrophage HD11 and immune tissues of chickens. Suppression of chicken HDPs expression may be an evasion strategy of C. jejuni for persistent colonization in chicken intestine by circumventing the chicken immune system.

Joint Application of Lactobacillus plantarum and Bacillus subtilis Improves Growth Performance, Immune Function and Intestinal Integrity in Weaned Piglets.

This study was conducted to explore the effects of the joint application of Lactobacillus plantarum and Bacillus subtilis on growth performance, immune function, antioxidant capacity, intestinal integrity, and gut microbiota composition in weaned piglets. The piglets were allocated randomly into 4 dietary groups, which were a control diet (NC), NC + 150 ppm mucilage sulfate (PC), and 3 additional diets containing 1 kg/t (LT), 1.5 kg/t (MT), or 2 kg/t (HT) mixture of Lactobacillus plantarum and Bacillus subtilis, respectively. Results showed that joint application of Lactobacillus plantarum and Bacillus subtilis increased ADFI and ADG of weaned piglets in d 14~28 and d 28~42 (p < 0.05), and decreased serum concentrations of DAO, IL-1β, TNF-α, and IL-2. The LT group increased jejunal and colonic sIgA contents compared with the PC group (p < 0.05). Groups of MT and HT increased colonic mRNA expression of host defense peptides and tight junction proteins compared with the NC and PC groups. The joint application of Lactobacillus plantarum and Bacillus subtilis increased the abundance of colonic Lactobacillus compared with NC and PC groups (p < 0.10). In conclusion, the joint application of Lactobacillus plantarum and Bacillus subtilis as an antibiotics alternative improved growth performance via promoting immune function and intestinal integrity of weaned piglets.

Temporal expression of avian [formula omitted] defensin 10 and cathelicidins in the yolk sac tissue of broiler and layer embryos

The yolk sac is a multifunctional organ, which not only participates in nutrient absorption, but also plays an important role in immune function. The objective of this study was to compare the mRNA abundance of avian β-defensin 10 (AvBD10) and 3 cathelicidins (CATH1, CATH2, and CATH3) in the yolk sac tissue (YST) of commercial broilers and white egg and brown egg commercial layers. AvBD10 and CATH mRNA abundance was analyzed using two-way ANOVA and Tukey's test, with P < 0.05 being considered significant. AvBD10 and CATH mRNA showed similar temporal expression patterns in the YST of both broiler and layers, with an increase from embryonic day (E) 7 to E9 through E13 followed by a decrease to day of hatch. AvBD10 mRNA showed a breed × age interaction with greater expression in the YST of both layers compared to broilers at E9 and E11. CATH1 mRNA was greater in the YST of brown egg layers than broilers. CATH2 mRNA showed a breed × age interaction, with greater expression in the YST of brown egg layers than broilers at E11. CATH3 mRNA showed no difference in the YST between layers and broilers. Because broilers and brown egg layers are genetically related, these results show that selection for production parameters (broiler vs. layer) and not genetic relatedness (white egg layer vs. brown egg layer and broilers) is the basis for the differences in AvBD10, CATH1, and CATH2 mRNA in the YST of broilers and layers. The yolk-free body weights of broiler embryos were greater than that of both brown and white egg layers from E9 to 17. One possible explanation is that the reduced expression of AvBD10, CATH1 and CATH2 mRNA in the YST of broilers compared to layers at E9 and 11 may be due to faster embryonic growth at the expense of host defense peptide expression in broilers compared to layers.

PSXI-14 Effects of Footpad Dermatitis on Gene Expression of Host Defense Peptides in Laying Hens

Abstract Footpad dermatitis (FPD) has a negative impact on the welfare and health production of poultry. The present study aimed to determine the effect of FPD on the expression of host defense peptides (HPDs). A total of fifty-four 55-week-old Hy-line brown hens housed individually in furnished cages were selected for investigating the mRNA expression of HPDs in foot, spleen, liver, and small intestine tissues. FPD score 0 group with normal foot, FPD score 1 with a small lesion of the foot pad epithelium and FPD score 2 with a larger lesion were divided by visual examination. The results showed that compared with group 0, hens from FPD group 1 and 2 had increased mRNA expression of HPDs of AvBD1, AvBD3, AvBD4, AvBD5, AvBD6, AvBD7, AvBD8, AvBD9, AvBD10, AvBD11, AvBD12, AvBD13, AvBD14 and LEAP2in foot and spleen tissues (P &amp;lt; 0.05). The relative expression of HPDs genes AvBD2, AvBD3, AvBD4, AvBD5, AvBD7, AvBD8, AvBD11, AvBD12, AvBD13, AvBD14, and LEAP2 in liver tissue increased, while the relative mRNA expression of AvBD6, AvBD9, and AvBD10 decreased (P &amp;lt; 0.05), and the expression levels of AvBD1, AvBD3, AvBD5, AvBD6, AvBD7, AvBD9, AvBD10, AvBD12, and LEAP2 in small intestinal tissue increased (P &amp;lt; 0.05), while the expression of AvBD2, AvBD8, and AvBD14 decreased (P &amp;lt; 0.05). Therefore, FPD can affect the expression of AVBDs in laying hens that the foot and spleen tissue had stronger responsiveness to the FPD. It is speculated that the increase in the expression of AVBDs on account of the possible infection with pathogenic bacteria. Then the increased HPDs participated in the host's defense responses and strengthened the immune regulation after FPD occurred.

Host Defense Peptides at the Ocular Surface: Roles in Health and Major Diseases, and Therapeutic Potentials.

Sight is arguably the most important sense in human. Being constantly exposed to the environmental stress, irritants and pathogens, the ocular surface – a specialized functional and anatomical unit composed of tear film, conjunctival and corneal epithelium, lacrimal glands, meibomian glands, and nasolacrimal drainage apparatus – serves as a crucial front-line defense of the eye. Host defense peptides (HDPs), also known as antimicrobial peptides, are evolutionarily conserved molecular components of innate immunity that are found in all classes of life. Since the first discovery of lysozyme in 1922, a wide range of HDPs have been identified at the ocular surface. In addition to their antimicrobial activity, HDPs are increasingly recognized for their wide array of biological functions, including anti-biofilm, immunomodulation, wound healing, and anti-cancer properties. In this review, we provide an updated review on: (1) spectrum and expression of HDPs at the ocular surface; (2) participation of HDPs in ocular surface diseases/conditions such as infectious keratitis, conjunctivitis, dry eye disease, keratoconus, allergic eye disease, rosacea keratitis, and post-ocular surgery; (3) HDPs that are currently in the development pipeline for treatment of ocular diseases and infections; and (4) future potential of HDP-based clinical pharmacotherapy for ocular diseases.

Epigenetic Regulation of Host Defense Peptide Synthesis: Synergy Between Histone Deacetylase Inhibitors and DNA/Histone Methyltransferase Inhibitors.

Host defense peptides (HDPs) are an integral part of the innate immune system acting as the first line of defense. Modulation of HDP synthesis has emerged as a promising host-directed approach to fight against infections. Inhibition of histone deacetylation or DNA methylation is known to enhance HDP gene expression. In this study, we explored a possible synergy in HDP gene induction between histone deacetylase inhibitors (HDACi) and DNA/histone methyltransferase inhibitors (DNMTi/HMTi). Two chicken macrophage cell lines were treated with structurally distinct HDACi, HMTi, or DNMTi individually or in combinations, followed by HDP gene expression analysis. Each epigenetic compound was found to be capable of inducing HDP expression. To our surprise, a combination of HDACi and HMTi or HDACi and DNMTi showed a strong synergy to induce the expressions of most HDP genes. The HDP-inducing synergy between butyrate, an HDACi, and BIX01294, an HMTi, were further verified in chicken peripheral blood mononuclear cells. Furthermore, tight junction proteins such as claudin 1 were also synergistically induced by HDACi and HMTi. Overall, we conclude that HDP genes are regulated by epigenetic modifications. Strategies to increase histone acetylation while reducing DNA or histone methylation exert a synergistic effect on HDP induction and, therefore, have potential for the control and prevention of infectious diseases.

Tibetan Pig-Derived Probiotic Lactobacillus amylovorus SLZX20-1 Improved Intestinal Function via Producing Enzymes and Regulating Intestinal Microflora.

The interaction between exogenous microorganisms and the host has received great attention, and finding new probiotics is always the way to improve the health of humans and animals. Lactobacillus amylovorus (L. amylovorus) is a kind of Lactobacillus that can efficiently utilize starch, as a food and feed additive, it has been widely used for mildew prevention and antibacterial, bacteriostasis, and enzyme production. Herein, a strain of L. amylovorus was isolated from the feces of Tibetan weaned piglets, named L. amylovorus SLZX20-1. Physiological and biochemical experiments in vitro confirmed that it had a fast growth rate and could produce a variety of enzymes, including α-galactosidase, β-galactosidase, α-glucosidase, β-glucosidase, and ferulic acid esterase. In addition, L. amylovorus SLZX20-1 exerted antibiotic effects on the growth of Salmonella typhimurium (S. typhimurium) SL1344, Citrobacter rodentium (C. rodentium) DBS100, Salmonella pullorum (S. pullorum) CVCC1791, Staphylococcus aureus (S. aureus) CVCC1882, Escherichia coli (E. coli) O157, E. coli K88, E. coli K99, and E. coli 987P, which are closely related to acid productivity, such as lactic acid and acetic acid. In vitro co-culture, L. amylovorus SLZX20-1 has shown the strong adhesion ability to intestinal porcine epithelial cells (IPEC-J2 cells) and activated IPEC-J2 cells with high expression of host defense peptides (HDPs), such as NK-Lysin, PEP2C, and PBD-1. In vivo experiment, via intragastric administration, L. amylovorus SLZX20-1 significantly improved the feed intake of mice, declined the crypt depth of jejunum and ileum, L. amylovorus SLZX20-1 changed the composition of intestinal microbes, especially at the level of colonic genus, the dominant genus was changed from Lactobacillus to S24-7, which indicated the change of intestinal carbohydrate nutrition. In conclusion, L. amylovorus SLZX20-1 showed strong probiotic characteristics, which met with the standard of probiotics and is worth further exploring its impacts on host health and its potential as a candidate strain of probiotics.

HPV infection alters vaginal microbiome through down-regulating host mucosal innate peptides used by Lactobacilli as amino acid sources

Despite the high prevalence of both cervico-vaginal human papillomavirus (HPV) infection and bacterial vaginosis (BV) worldwide, their causal relationship remains unclear. While BV has been presumed to be a risk factor for HPV acquisition and related carcinogenesis for a long time, here, supported by both a large retrospective follow-up study (n = 6,085) and extensive in vivo data using the K14-HPV16 transgenic mouse model, we report a novel blueprint in which the opposite association also exists. Mechanistically, by interacting with several core members (NEMO, CK1 and β-TrCP) of both NF-κB and Wnt/β-catenin signaling pathways, we show that HPV E7 oncoprotein greatly inhibits host defense peptide expression. Physiologically secreted by the squamous mucosa lining the lower female genital tract, we demonstrate that some of these latter are fundamental factors governing host-microbial interactions. More specifically, several innate molecules down-regulated in case of HPV infection are hydrolyzed, internalized and used by the predominant Lactobacillus species as amino acid source sustaining their growth/survival. Collectively, this study reveals a new viral immune evasion strategy which, by its persistent/negative impact on lactic acid bacteria, ultimately causes the dysbiosis of vaginal microbiota.

Synergistic Induction of Chicken Antimicrobial Host Defense Peptide Gene Expression by Butyrate and Sugars.

Antimicrobial resistance is a major concern to public health demanding effective alternative strategies to disease control and prevention. Modulation of endogenous host defense peptide (HDP) synthesis has emerged as a promising antibiotic alternative approach. This study investigated a potential synergy between sugars and butyrate in inducing HDP gene expression in chickens. Our results revealed that sugars differentially regulated HDP expression in both gene- and sugar-specific manners in chicken HD11 macrophage cells. Among eight mono- and disaccharides tested, all were potent inducers of avian β-defensin 9 (AvBD9) gene (p<0.05), but only galactose, trehalose, and lactose obviously upregulated cathelicidin-B1 (CATHB1) gene expression. The expression of AvBD14 gene, on the other hand, was minimally influenced by sugars. Moreover, all sugars exhibited a strong synergy with butyrate in enhancing AvBD9 expression, while only galactose, trehalose, and lactose were synergistic with butyrate in CATHB1 induction. No synergy in AvBD14 induction was observed between sugars and butyrate. Although lactose augmented the expression of nearly all HDP genes, its synergy with butyrate was only seen with several, but not all, HDP genes. Mucin-2 gene was also synergistically induced by a combination of lactose and butyrate. Furthermore, lactose synergized with butyrate to induce AvBD9 expression in chicken jejunal explants (p<0.05). Mechanistically, hyper-acetylation of histones was observed in response to both butyrate and lactose, relative to individual compounds. Mitogen-activated protein kinase, NF-κB, and cyclic adenosine monophosphate signaling pathways were also found to be involved in butyrate- and lactose-mediated synergy in AvBD9 induction. Collectively, a combination of butyrate and a sugar with both HDP-inducing and barrier protective activities holds the promise to be developed as an alternative to antibiotics for disease control and prevention.

Evaluation of Potential Probiotic Properties of Lactobacillus and Bacillus Strains Derived from Various Sources for Their Potential Use in Swine Feeding.

Beneficial effects of probiotics are relevant to the various potential properties of individual strains, and they may also relate to the original sources of the probiotic strains. This study aimed to characterize the potential probiotic properties of the strains originating from various sources for probiotics use in swine feeding. A total of 9 potential probiotic strains, seven lactobacilli and 2 bacilli, were examined for antimicrobial production against swine pathogens, adhesion and anti-adhesion of potential probiotic strains to IPEC-J2 cells, aggregation ability, host defense peptide expression, and hemolytic assay. The results highlight that all strains derived from different sources could exhibit probiotic properties, although different abilities were observed. L. rhamnosus SD11 exhibited the highest inhibitory effect against all pathogens compared to other strains. Bacillus licheniformis KMP-9, B. subtilis KMP-N004, and L. fermentum SD7 gave the highest internalization and that related to high abilities of exclusion, competition, and displacement inhibition to pathogens. Such strains also gave a higher co-aggregation to all pathogens compared to other potential probiotic strains. L. rhamnosus GG, L. fermentum SD7, L. rhamnosus SD4, and B. subtilis KMP-N004 had significantly higher pBD-2 mRNA expression than other strains. None of potential probiotic strains showed hemolytic activity. In conclusion, the strains derived from either humans or animals possessed desirable probiotic properties including inhibition against porcine pathogens, adhesion capacity to porcine enterocytes, anti-adhesion pathogens to porcine enterocytes, and modulated innate immunity. Results indicate that these probiotic strains may be good candidates for use in swine feeding to reduce the risk of infection.

Long-term exposure to particulate matter from air pollution alters airway β-defensin-3 and -4 and cathelicidin host defense peptides production in a murine model

Epidemiological studies have associated long-term exposure to environmental air pollution particulate matter (PM) with the development of diverse health problems. They include infectious respiratory diseases related to the deregulation of some innate immune response mechanisms, such as the host defense peptides' expression. Herein, we evaluated in BALB/c mice the effect of long-standing exposure (60 days) to urban-PM from the south of Mexico City, with aerodynamic diameters below 2.5 μm (PM2.5) and 10 μm (PM10) on the lung's gene expression and production of three host defense peptides (HDPs); murine beta-defensin-3, -4 (mBD-3, mBD-4) and cathelin-related antimicrobial peptide (CRAMP). We also evaluated mRNA levels of Il1b and Il10, two cytokines related to the expression of host defense peptides. Exposure to PM2.5 and PM10 differentially induced lung inflammation, being PM2.5, which caused higher inflammation levels, probably associated with a differential deposition on the airways, that facilitate the interaction with alveolar macrophages. Inflammation levels were associated with an early upregulation of the three HDPs assessed and an increment in Il1b mRNA levels. Interestingly, after 28 days of exposure, Il10 mRNA upregulation was observed and was associated with the downregulation of HDPs and Il1b mRNA levels. The upregulation of Il10 mRNA and suppression of HDPs might facilitate microbial colonization and the development of diseases associated with long-term exposure to PM.

SPROUTY2 INHIBITS EXPRESSION OF THE HOST DEFENSE PEPTIDE RELMβ IN THE COLONIC EPITHELIUM

Abstract Background Sprouty2 (Spry2) is an inflammation-responsive regulator of intracellular signaling that is suppressed in colitis. Loss of Spry2 levels leads to increased colonic tuft and goblet cell numbers; cell types protective against injury and infection. In particular, goblet cells produce host defense peptides (HDPs) which can enhance barrier effectiveness and limit pathogen colonization. While the activity of these cells is regulated and responsive to environmental cues, the precise mechanisms driving functional changes in goblet cells are largely unknown. Here, we tested the role of Spry2 in mediating colonic goblet cell function and HDP production. Hypothesis Spry2 regulates HDP production in colonic goblet cells and bacterial levels. Methods Distal colons from mice with an intestinal epithelial-specific Spry2 deletion (Spry2flox/flox;Villin-Cre, Spry2KOIE) and littermate controls were collected at 8 w of age, and analyzed by bulk and single-cell RNA sequencing. Fecal pellets from the distal colon were 16S sequenced to assess changes in microbial populations. Changes in HDP expression were assessed at the bulk tissue level and within individual cellular populations. Results Expression of the host defense peptide RELMβ, but not RELMα, RELMγ, or Resistin, was increased 15-fold (p&amp;lt;0.05) in Spry2KOIE mice in bulk RNA sequencing analysis. Single cell RNA sequencing showed RELMβ was highly enriched in Reg4+/Spink4+ putative crypt base goblet cells of both control and Spry2KOIE mice. Quantification of these cells by single-cell RNA sequencing revealed no significant difference in Reg4+/Spink4+ cell numbers in Spry2KOIE mice versus wild-type, suggesting increased RELMβ is due to goblet cell activation and not goblet cell population shifts. Taxonomic analysis of feces from 2 litters of mice showed a decreased abundance of Eggerthella spp. (reduced 61%, p&amp;lt;0.05) and Streptococcus spp. (reduced 50%, p&amp;lt;0.05), genera that have been identified in patients with inflammatory bowel disease. Conclusion Loss of epithelial Spry2 drives HDP production in the colon and alterations in the colonic microbiota. Impairment of this mechanism may impact colitis severity.

Profiles of expression pattern and tissue distribution of host defense peptides genes in different chicken (Gallus gallus) breeds related to body weight.

Host defense peptides (HDPs) are an important first line of defense with antimicrobial and immunomodulatory properties. Selection for increased body weight is hypothesized to be related to reduced immune response. We studied the relationships among body weight, age, and the HDP expression patterns in intestine and immune organs. We used chickens with marked differences of body sizes. The non-selected Daweishan mini chickens showed the highest indexes of immune organs and the lowest concentrations of the plasma immune parameters C3, C4, IgA, and IgY, while the commercial Avian broiler showed the opposite results. The Daweishan mini chickens showed the highest mRNA expressions of HDP genes in small intestine followed by the semi-selected Wuding chickens. Compared with local breeds, broiler chickens showed higher mRNA expression of HDP genes in spleen, thymus, and bursa. Body weight and HDP expression levels were negatively correlated in the intestine and positively in the immune organs. Our results indicated that the HDP immune regulatory roles in small intestine acted as first line of defense in innate immunity in local breeds, and as an adaptive immunity in broiler chickens. Selection was associated with different expression expressions of HDP genes in breed-, age-, and organ-specific manners.

Does an Apple a Day Also Keep the Microbes Away? The Interplay Between Diet, Microbiota, and Host Defense Peptides at the Intestinal Mucosal Barrier

A crucial mechanism of intestinal defense includes the production and secretion of host defense peptides (HDPs). HDPs control pathogens and commensals at the intestinal interface by direct killing, by sequestering vital ions, or by causing bacterial cells to aggregate in the mucus layer. Accordingly, the combined activity of various HDPs neutralizes gut bacteria before reaching the mucosa and thus helps to maintain the homeostatic balance between the host and its microbes at the mucosal barrier. Defects in the mucosal barrier have been associated with various diseases that are on the rise in the Western world. These include metabolic diseases, such as obesity and type 2 diabetes, and inflammatory intestinal disorders, including ulcerative colitis and Crohn's disease, the two major entities of inflammatory bowel disease. While the etiology of these diseases is multifactorial, highly processed Western-style diet (WSD) that is rich in carbohydrates and fat and low in dietary fiber content, is considered to be a contributing lifestyle factor. As such, WSD does not only profoundly affect the resident microbes in the intestine, but can also directly alter HDP function, thereby potentially contributing to intestinal mucosal barrier dysfunction. In this review we aim to decipher the complex interaction between diet, microbiota, and HDPs. We discuss how HDP expression can be modulated by specific microbes and their metabolites as well as by dietary factors, including fibers, lipids, polyphenols and vitamins. We identify several dietary compounds that lead to reduced HDP function, but also factors that stimulate HDP production in the intestine. Furthermore, we argue that the effect of HDPs against commensal bacteria has been understudied when compared to pathogens, and that local environmental conditions also need to be considered. In addition, we discuss the known molecular mechanisms behind HDP modulation. We believe that a better understanding of the diet-microbiota-HDP interdependence will provide insights into factors underlying modern diseases and will help to identify potential dietary interventions or probiotic supplementation that can promote HDP-mediated intestinal barrier function in the Western gut.

Sodium Butyrate Protects the Intestinal Barrier by Modulating Intestinal Host Defense Peptide Expression and Gut Microbiota after a Challenge with Deoxynivalenol in Weaned Piglets.

This study aims to determine whether sodium butyrate (SB) could antagonize deoxynivalenol (DON)-induced intestinal epithelial dysfunction. In a four-week feeding trial, twenty-eight barrows were randomly divided into four treatments: (1) uncontaminated basal diet (control); (2) 4 mg/kg DON-contaminated diet (DON); (3) basal diet supplemented with 0.2% SB (SB); and (4) 4 mg/kg DON + 0.2% SB (DON + SB). A decrease in performance was observed in DON-exposed animals, which was prevented by the dietary SB supplementation. DON exposure also depressed the expression of host defense peptides (HDPs) in the intestine, impaired the intestinal barrier integrity, and disturbed the gut microbiota homeostasis. These alterations induced by DON were attenuated by SB supplementation. The supplementation of 0.2% SB ameliorated the adverse effects of DON on the liver in terms of hepatic lesions as well as serum concentrations of alkaline phosphatase and aspartate aminotransferase. In IPEC-J2 cells, pretreatment with SB alleviated the DON-induced decreased cell viability. Additionally, the NOD2/caspase-12 pathway participated in the alleviation of SB on DON-induced diminished HDP expression. Taken together, these data demonstrated that SB protected piglets from DON-induced intestinal barrier dysfunction potentially through stimulation of intestinal HDP assembly and regulation in gut microbiota.