Β-defensin Research Articles

P44 Upregulation of beta defensin in human epidermal organoids treated with combinations of IL17 and IL23

Abstract Psoriasis is a chronic inflammatory skin disease characterized by the hyperproliferation of keratinocytes and an aberrant immune response. The pro-inflammatory cytokines IL-17 (a, e, f) and IL-23 play crucial roles in the pathogenesis of psoriasis. Human epidermal organoids (HEOs) are an in vitro model that recapitulates the epidermis and a useful tool for studying epidermal biology and disease mechanisms. HEOs were treated with various combinations of cytokines, including IL-17a, IL-17e, IL-17f, and IL-23. Following treatment, quantitative PCR (qPCR) and immunohistochemistry (IHC) were performed to assess the expression levels of beta-defensin, an antimicrobial peptide implicated in psoriasis. The qPCR analysis revealed significant upregulation of beta defensin in HEOs treated with the following cytokine combinations: IL-17a alone, IL-17f alone, IL-17a+e, IL-17a+f, IL-17a+IL-23, IL-17a+e+f, IL-17a+e+IL-23, IL-17a+f+IL-23, and IL-17a+e+f+IL-23. Conditions lacking IL-17a exhibited markedly lower beta-defensin expression, indicating the critical role of IL-17a in this process. However, IL-17f alone induced a moderate upregulation of beta-defensin. The study demonstrates that IL-17a is a pivotal cytokine in inducing beta-defensin expression in HEOs, with IL-17f also contributing to a lesser extent. The significant upregulation of beta-defensin in response to these cytokines suggests a key role for beta-defensin in the pathophysiology of psoriasis, potentially linking antimicrobial peptide activity to disease severity and progression.

FC14 Modulation of the IL23/IL17 axis beyond IL-17A drives response to risankizumab in palmoplantar psoriasis

Abstract Palmoplantar psoriasis (PPPsO) is a variant of plaque psoriasis (PsO) that is characterized by difficult-to-treat lesions on the hands and feet and is associated with substantial physical disability and discomfort. A better understanding of pathogenic disease drivers is critical to ensuring patients receive treatments that will drive high levels of efficacy. Data from non-head-to-head randomized clinical trials suggest that IL-23 blockade may be more efficacious than IL17A blockade in PPPsO. Here, we evaluated changes in downstream mediators of the IL-23/IL-17 axis following treatment with risankizumab, an interleukin (IL)-23 inhibitor approved for the treatment of PsO and demonstrate that broad coverage of the IL-23 pathway beyond IL-17A may be required to achieve >90% improvement in clinical score in PPPsO. IMMprint (NCT04713592) is an ongoing Phase 3b, multicentre, randomized, double-blind, placebo-controlled study for patients with PPPsO. Patients were randomized 1:1 to receive either blinded 150 mg risankizumab or placebo at Weeks 0 and 4. At Week 16 all patients entered the open-label extension and received risankizumab at Week 16 and every 12 weeks thereafter. Whole blood was collected (N = 35 in each arm) at baseline and Weeks 4, 16, and 52. Serum cytokine levels of IL-17A and IL-17C were assessed by proximity extension assay and IL-22 and beta-defensin 2 (BD-2) were assessed by high sensitivity individual assays. Efficacy at Week 16 was assessed by psoriasis area and severity index (PASI) and palmoplantar PASI (PPASI). Response to risankizumab was defined as achieving the PPASI 90 endpoint. A total of 13 patients receiving risankizumab and 2 patients receiving placebo achieved PPASI 90 at Week 16. Patients that achieved PPASI 90 with risankizumab had a significant reduction in serum levels of IL-22 (pro-inflammatory cytokine), BD-2 (host defence), and IL-17C (amplification of epithelial inflammation) as early as 4 weeks after treatment, continuing through Week 52, compared with patients that did not achieve PPASI 90. Further, a positive correlation between reduction in IL-22, BD-2, and IL-17C and PASI and PPASI scores was observed at Week 16 for patients receiving risankizumab but not placebo. No association was observed between other inflammatory molecules, including IL-17A, and patient achievement of PPASI 90. The data suggest that the IL-23/IL17 axis beyond just IL-17A is important for driving disease pathology in PPPsO.

P74 Guselkumab clinical super-response is associated with faster normalization of blood and skin pathology: data from the Phase 3b GUIDE trial

Abstract The ongoing GUIDE trial (NCT03818035) examines the clinical and immunological impact of an extended dosing interval of guselkumab [an interleukin (IL)-23p19 subunit inhibitor] in patients with moderate-to-severe plaque-type psoriasis. A mechanistic substudy was conducted to explore the molecular differences in the skin and blood of ‘super-responder’ (SRe) and non-SRe patients. SRes were defined as patients achieving a psoriasis area and severity index (PASI) = 0 at both Week [W]20 and W28 following guselkumab treatment. RNA-sequencing was performed on non-lesional (at W0) and lesional skin samples (at W0, W4, W28, and W68) along with serum analyses of matching timepoints. Published gene sets related to psoriasis skin and cytokine-stimulated keratinocytes were used for gene set variation analysis. At W0, ∼1100 protein-coding transcripts were upregulated in lesional vs. non-lesional skin, and ∼3000 protein-coding transcripts were downregulated in lesional vs. non-lesional skin. Following treatment with guselkumab, robust pharmacodynamic changes were observed for immune- and psoriasis inflammation-related gene signatures. Faster changes were observed in SRes vs. non-SRes. Signatures related to psoriasis skin inflammation, IL-17-stimulated keratinocyte response, T-helper (Th)17 cells, and many keratinocyte subtypes (differentiated, proliferating and inflamed keratinocytes) were more significantly reduced in SRes vs. non-SRes as early as W4. By W28, all signatures that were upregulated at baseline in lesional vs. non-lesional skin were normalized in SRes, including molecular scar skin and Th17 pathway signatures. In contrast, among non-SRes, molecular scar and immune-cell-regulated signatures (including macrophage, Th22, mucosal-associated invariant T cells, and CD4+ T cells) were <75% normalized at W28. In both SRes and non-SRes, signatures initially downregulated in lesional vs. non-lesional skin at W0, including those for adipocytes, smooth muscles, meta-analysis-derived psoriasis downregulated genes, and IL-17-stimulated KC downregulated genes, represent the few signatures that did not return to non-lesional skin levels after 68 weeks of guselkumab treatment. Consistent with the effect of guselkumab on inflammatory processes in the skin as observed via RNA sequencing, guselkumab treatment decreased the serum levels of several inflammatory mediators as early as W4. The levels of IL-17A, IL-17F and IL-22 were comparable in SRes and non-SRes, while the reduction in beta defensin-2 level was more pronounced in SRes at W4 and W28. These reductions were maintained to W68. In summary, molecular analyses of SRes and non-SRes in the GUIDE clinical trial show kinetic and quantitative differences in the response to guselkumab treatment.

Effects of Cigarette Smoking on Host Defense Peptides in Saliva of Patients with Periodontitis

Background: Smoking cigarettes is linked to a number of diseases (systemic and oral), and affect both innate and adaptive immunity. Host defense peptides, often known as antimicrobial peptides (AMPs), are a component of the innate immune system and regarded as a group of chemically active oligopeptides that are poisonous to harmful microbes. Aims: The goal of this research was to shed light on the effect of cigarette smoking on the production of host defense peptides in periodontitis patients. Methods and Materials: Eighty-five participants were included in this study. Periodontal health assessed by: PLI, GI, BOP, PPD and CAL. Saliva samples were collected from all subjects. ELISA was carried out to estimate the levels of beta defensin and cathelicidin. Statistical analysis was performed using SPSS version 24. The ANOVA parametric test was used to determine and detect the differences between independent study groups, and the Tukey honestly significant difference (HSD)/post hoc test was performed to determine if there is any statistical connection between two data sets (intergroup comparison). Results: The study findings showed both smokers and non smokers with periodontitis had considerably greater levels of beta defensin and cathelicidin than healthy controls. There was statistically significant reduction in beta defensin between smokers and non smokers (p<0.001); whereas no significant differences found between smokers and non smokers regarding cathelicidin level. Conclusions: These findings suggested that, in the context of periodontal diseases, smoking can affect the protein levels of beta defensin but not the levels of cathelicidin

Design of a novel multi-epitope vaccine candidate against Yersinia pestis using advanced immunoinformatics approaches: An in silico study

Yersinia pestis is the perilous pandemics that occurred in Asia and Europe. The bacterium has shown drug resistance that can cause the future pandemic and destroy the drug treatment against plague. As known, effective therapeutics such as designing potent vaccine that can aid world to protect against plague. The immunoinformatics approaches was implemented via different server. The 4 potent antigens (F1 capsule, LcrV, OmpA, and PH6) were listed as essential protein target for creating the multi-epitope vaccine. These targets were selected for designing multi-epitope vaccine that predicted the CTL and HTL epitopes. The vaccine construct included different linkers such as EAAAK, AAY, GPGPG, and SSL that an adjuvant (Beta defensin-3) inserted at N-terminal of vaccine. The computational physiochemical properties and other immunological analysis showed stable, soluble, antigen, non-allergen, and non-toxin. The molecular docking confirmed the stable binding and good interaction and the iMODS server showed the stable binding. Furthermore, computational immune simulation of multi-epitope vaccine showed that vaccine can stimulate adaptive and innate responses after second doses. In this study, the vaccine designed for Y. pestis that in future require immunological examination to unveil real efficiency.

КЛИНИЧЕСКО-ИММУНОЛОГИЧЕСКИЕ АСПЕКТЫ ПРИ ХИРУРГИЧЕСКОМ ЛЕЧЕНИИ ПАЦИЕНТОВ С ГИПЕРКЕРАТОЗАМИ СЛИЗИСТОЙ ОБОЛОЧКИ РТА

Relevance. The relevance of the treatment of oral mucosa diseases is determined by their prevalence, propensity to progression and difficulties in early diagnosis. The article presents the results of surgical treatment of this category of patients using laser radiation with a wavelength of 1940 nm and determination of gene expression of immunologic factors. Purpose. To increase the efficiency of surgical treatment of oral mucosa hyperkeratosis by using the fiber laser LSP – «IRE-Polus». Materials and methods. Surgical treatment was performed on 30 patients (100%) with clinical diagnosis K13.2 Leukoplakia and other disturbances of oral epithelium, including tongue (25 patients) and L43.0 Lichen ruber planus hypertrophicus (5 patients). All patients were randomly divided into 2 groups: in the 1st group lesions were excised by fiber laser LSP – «IRE-Polus», in the 2nd group excision was performed by scalpel. Sampling for gene expression study of defensin beta 1 and IL-28B genes was performed after surgical intervention in all patients. Statistical analysis was carried out using the Mann–Whitney test. Results. The use of laser alteration leads to a decrease in pain intensity, collateral edema, and the timing of postoperative wound epithelialization. In the expression study of immunologic factors, there was no statistically significant difference in the expression of defensin beta 1 gene in both study groups, while an association was found between a high level of IL-28B gene expression and the development of inflammation when using the traditional method. Conclusion. The use of fiber lasers contributes to the improvement of treatment efficiency in patients with hyperkeratosis of the oral mucosa. When examining patients with hyperkeratosis, a significant 2.2-fold increase in IL-28B gene expression was found in the group of patients where the traditional method of treatment was applied (p < 0.05).

Human β-Defensin 2 as a Link between EBV Infection and Multiple Sclerosis

Multiple sclerosis (MS) is the result of a complex interaction between environmental factors and genetic predisposition. Epstein-Barr virus (EBV) is a significant environmental risk factor associated with MS. Human- beta defensin 2 (HβD-2) is a crucial innate immune response proteint that as an inflammatory marker protects humans from pathogens invading the body like viruses. Therefore, a case-control study was conducted on 90 subjected (48 patients with MS and 42 healthy controls) to examine the role of HβD-2 in EBV viral load infection and MS patients. An enzyme-linked immunosorbentassay kit was used to measure HβD-2 levels in serum. The viral load of EBV was determined using a real-time PCR. The findings revealed that median HβD-2 levels in patients were significantly lower than in controls. (96.62 vs.124.8 ng/mL; p < 0.01). According to the receiver operating characteristic curve analysis, HβD-2 is good predictor for MS disease (area under the curve=0.739; p < 0.001). EBV frequency was lower in MS patients compared to the controls, although difference was not statistically significant (25 vs. 38.1%; p = 0.181).On other hand, the median (EBV) load was substantially greater in patients than in healthy controls ( 8.55 vs. 1.4 DNA copy/100 cells). This study concluded that HβD-2 levels showed no significant differences in each age group, sex, Expanded Disability Status Scale (EDSS) of multiple sclerosis patients, rather showed an inverse significant correlation (correlation coefficient = - 0.432) with EBV load, which protect against human viral infections.

Gene expression responses of CF airway epithelial cells exposed to elexacaftor/tezacaftor/ivacaftor suggest benefits beyond improved CFTR channel function.

The combination of elexacaftor/tezacaftor/ivacaftor (ETI, Trikafta) reverses the primary defect in cystic fibrosis (CF) by improving CFTR-mediated Cl- and HCO3- secretion by airway epithelial cells (AECs), leading to improved lung function and less frequent exacerbations and hospitalizations. However, studies have shown that CFTR modulators like ivacaftor, a component of ETI, have numerous effects on CF cells beyond improved CFTR channel function. Because little is known about the effect of ETI on CF AEC gene expression, we exposed primary human AEC to ETI for 48 h and interrogated the transcriptome by RNA-seq and qPCR. ETI increased CFTR Cl- secretion, and defensin gene expression (DEFB1), an observation consistent with reports of decreased bacterial burden in the lungs of people with CF (pwCF). ETI decreased MMP10 and MMP12 gene expression, suggesting that ETI may reduce proteolytic-induced lung destruction in CF. ETI also reduced the expression of the stress response gene heme oxygenase (HMOX1). qPCR analysis confirmed DEFB1, HMOX1, MMP10, and MMP12 gene expression results observed by RNA-seq. Gene pathway analysis revealed that ETI decreased inflammatory signaling, cellular proliferation, and MHC class II antigen presentation. Collectively, these findings suggest that the clinical observation that ETI reduces lung infections in pwCF is related in part to drug-induced increases in DEFB1 and that ETI may reduce lung damage by reducing MMP10 and MMP12 gene expression. Moreover, pathway analysis also identified several other genes responsible for the ETI-induced reduction in inflammation observed in pwCF.NEW & NOTEWORTHY Gene expression responses by CF AECs exposed to ETI suggest that in addition to improving CFTR channel function, ETI is likely to enhance resistance to bacterial infection by increasing levels of beta-defensin 1 (hBD-1). ETI may also reduce lung damage by suppressing MMP10 and MMP12 and reduce airway inflammation by repressing proinflammatory cytokine secretion by CF AECs.

Synergistic role of Mycobacterium indicus pranii and human beta Defensin-2 as adjunctive therapy against Mycobacterium tuberculosis

Host-directed therapies (HDT) via modulation of specific host responses like inflammation can limit mycobacterial infection. HDTs could be included in current TB therapy as an adjunct to increase bacterial clearance and limit tissue damage to control spread. Individually, Mycobacterium indicus pranii (MIP) and human beta defensin-2 (hBD-2) are promising therapies for tuberculosis (TB). They can directly target the TB bacilli and enhance cell-mediated immune responses, which is limiting with conventional drugs. Therefore, our study investigated the combined application of MIP and hBD-2 to evaluate their efficacy in clearing infections caused by Mycobacterium smegmatis (M.smeg) and Mycobacterium tuberculosis (M.tb) (both avirulent; H37Ra and virulent strain; H37Rv) in THP-1 cells and human monocyte-derived macrophages (MDMs). A strong pro-inflammatory response was observed against the combination of MIP and hBD-2 which also correlated with a significant reduction in the bacterial load. This combination further showed protection against M.tb by enhancing pyroptosis in the infected cells. The study suggests the combined use of these potent immunomodulators, which could be employed as an effective mode of therapy as adjuvants against mycobacterial infections after validation in a suitable animal model.

Polymorphic variation of the DEFB1 gene might contribute to the development of ankylosing spondylitis: a preliminary study.

Ankylosing spondylitis (AS) is an inflammatory disease that affects the spine and can cause peripheral arthritis, enthesitis, and dactylitis, as well as extra-articular manifestations such as uveitis and inflammatory bowel disease. β-Defensins are antimicrobial peptides involved in the activation and regulation of several immune cell types that may influence the inflammatory response in AS. The aim was to analyze the association and interaction of two functional variants of the DEFB1 gene in AS patients, and their role with inflammatory markers. The rs11362 and rs1800972 variants were genotyped using TaqMan probes in Mexican AS patients and controls. C-reactive protein (CRP) levels and erythrocyte sedimentation rate (ESR) were quantified. SPSS software was used for statistical analysis and multifactor dimensionality reduction (MDR) for interactions. The AA and GG genotypes were associated with AS risk in the age- and sex-adjusted model (OR = 6.89, P = 0.008 and OR = 3.43, P = 0.046, respectively); furthermore, the A-G haplotype showed a significant association with AS risk (OR = 2.94, P = 0.012). ESR and CRP were elevated in carriers of the AA genotype compared to the GA and GG genotypes of the rs11362 variant (20.89 ± 9.78 vs. 5.63 ± 4.61 and 4.10 ± 2.65mm/h, P < 0.0001; and 10.92 ± 14.09 vs. 2.14 ± 2.02 and 2.15 ± 2.13mg/L, P < 0.001, respectively). Using the MDR method, strong interactions of the rs11362 variant with sex were identified in the adjusted and unadjusted models. These results suggest that the DEFB1 gene may play a key role in AS pathogenesis.

Pioneering Clinical Investigation: Beta Defensin Expression in Patients of Squamous Cell Carcinoma of Oral Cavity as a Forward-Looking Validation for Molecular Treatment

Pioneering Clinical Investigation: Beta Defensin Expression in Patients of Squamous Cell Carcinoma of Oral Cavity as a Forward-Looking Validation for Molecular Treatment

Features of the expression of genes of antimicrobial peptides and the microbiome at the level of the mucous membrane of the upper respiratory tract during aging

Today, the proportion of elderly and senile people is steadily growing throughout the world. The most important factors in the first line of immune defense of the mucous membranes of the upper respiratory tract are β-defensins, which are a group of secretory proteins with antimicrobial activity. The aim of this study was to study the expression of genes for antimicrobial peptides β-defensins and the composition of the microbiome of the mucous membranes of the upper respiratory tract in elderly people and long-livers with various aging phenotypes.The main study group included 67 centenarians and 49 elderly people, who were further divided into two subgroups depending on the course of aging (pathological and successful aging). Nucleic acids were isolated from nasopharyngeal scrapings and the expression levels of the DEFB1 and DEFB4 genes were determined using real-time polymerase chain reaction. The composition of the microbiota in nasopharyngeal swabs was determined by MALDI-TOF mass spectrometry.In analyzing the expression of the DEFB1 gene in elderly people and centenarians with successful and pathological aging phenotypes, no difference was revealed between the groups. Expression of the DEFB4 gene was increased in centenarians with pathological aging compared to centenarians with successful aging and in the elderly group. Excessive production of antimicrobial peptides is dual in nature; on the one hand, they provide the first line of defense against microorganisms, and on the other, they are cytotoxic to their own cells. An increase in the expression of the DEFB4 gene during aging may be due to an increase in the number of pathogen-associated molecular patterns, which can be one’s own microbiota and/or components of microbial metabolism. Analysis of the microbiota composition showed an increase in biodiversity in individuals with a successful aging phenotype compared to a pathological phenotype. Particular attention is paid to Staphylococcus spp., the species composition of which depends on the aging phenotype. In the pathological aging group, the frequency of St. aureus colonization is significantly higher than in the successful aging group.Thus, overexpression of the DEFB4 gene and changes in the composition of the microbiota of the mucous membranes of the upper respiratory tract may be one of the mechanisms explaining the increased susceptibility to infections in various aging phenotypes.

The role of epigenetic factors in childhood atopic dermatitis

Atopic dermatitis (AD) is a genetically determined chronic inflammatory skin disease characterized by itching, relapsing course, age-related features of the lesion morphology and localization. The etiological factors contributing to the development of AD are: allergic reactions, inflammatory reactions in the skin, leading to a barrier disfunction, the affect and interaction of genetic and environmental factors. Epigenetic mechanisms also play a key role in immune regulation. The aim of this work is to study the genome-wide methylation profile in the skin of children with AD in the acute stage, as well as to determine changes in the expression level of immune response genes associated with common signaling pathways during therapy. Whole-genome sequencing was conducted to examine methylation in the skin samples from AD patients and controls. The stages of bioinformatics data processing were carried out. Changes in the expression levels of the TLR2, TLR9, IL4, IL13, CAMP, and DEFB1 genes before and after treatment were carried out using RT-PCR-RT on samples of mononuclear blood cells. When comparing samples from patients with AD and healthy children, changes were shown in the methylation of 2364 regions of genomic DNA with their corresponding specific genes. Among the identified genes, those related to processes associated with the pathogenesis of AD were selected. At the next stage, changes in the expression of the described immunity factors (TLR2, TLR9, IL-4, IL-13, DEFB1, CAMP) were assessed over time according to the type of treatment. For this purpose, patients were divided into 2 groups depending on the prescription of systemic therapy. Three months after treatment, the level of expression for all studied immune factors decreased. However, a significant decrease in expression for almost all parameters (except for the DEFB1 gene) was recorded in the group in which systemic therapy (dupilumab) was used in addition to topical treatment. At the same time, the SCORAD index after treatment improved by an average of 63% and amounted to 17±6.0 points. Thus, systemic therapy aimed at suppressing signaling of IL-4 and IL-13 cytokines indirectly leads to normalization of the expression levels of innate immune factors. Studying the regulation of molecular genetic mechanisms of immunity involved in the processes of inflammation in AD helps to clarify the immunopathogenesis of this disease, determine diagnostic markers and targets for drug therapy.

Targeting Polyprotein to Design Potential Multiepitope Vaccine against Omsk Hemorrhagic Fever Virus (OHFV) by Evaluating Allergenicity, Antigenicity, and Toxicity Using Immunoinformatic Approaches.

Omsk Hemorrhagic Fever Virus (OHFV) is an RNA virus with a single-stranded, positive-sense genome. It is classified under the Flaviviridae family. The genome of this virus is 98% similar to the Alkhurma hemorrhagic fever virus (AHFV), which belongs to the same family. Cases of the virus have been reported in various regions of Saudi Arabia. Both OHFV and AHFV have similarities in pathogenic polyprotein targets. No effective and licensed vaccines are available to manage OHFV infections. Therefore, an effective and safe vaccine is required that can activate protective immunity against OHFV. The current study aimed to design a multiepitope subunit vaccine against the OHFV utilizing several immunoinformatic tools. The polyprotein of OHFV was selected and potent antigenic, non-allergenic, and nontoxic cytotoxic T-lymphocyte (CTL), helper T-lymphocyte (HTL), and linear B-lymphocyte (LBL) epitopes were chosen. After screening, eight (8) CTL, five (5) HTL, and six (6) B cell epitopes were joined with each other using different linkers. Adjuvant human beta defensin-2 was also linked to the epitopes to increase vaccine antigenic and immunogenic efficiency. The designed vaccine was docked with Toll-like receptor 4 (TLR4) as it activates and induces primary and secondary immune responses against OHFV. Codon optimization was carried out, which resulted in a CAI value of 0.99 and 53.4% GC contents. In addition, the construct was blindly docked to the TLR4 immune receptor and subjected to conformational dynamics simulation analysis to interpret the intricate affinity and comprehend the time-dependent behavior. Moreover, it was predicted that immune responses to the developed vaccine construct reported formation of strong humoral and cellular immune cells. Therefore, the proposed vaccine may be considered in experimental assays to combat OHFV infections. Laboratory experiments for the above predictions are essential in order to evaluate the effectiveness, safety, and protective properties of the subject in question.

Enhanced Antimicrobial Peptide Response Following Bacillus Calmette-Guerin Vaccination in Elderly Individuals.

Antimicrobial peptides are an important component of host defense against Mycobacterium tuberculosis. However, the ability of BCG to induce AMPs as part of its mechanism of action has not been investigated in detail. We investigated the impact of Bacillus Calmette-Guerin (BCG) vaccination on circulating plasma levels and TB-antigen stimulated plasma levels of AMPs in a healthy elderly population. We assessed the association of AMPs, including Human Beta Defensin 2 (HBD-2), Human Neutrophil Peptide 1-3 (HNP1-3), Granulysin, and Cathelicidin (LL37), in circulating plasma and TB-antigen stimulated plasma (using IGRA supernatants) at baseline (pre-vaccination) and at Month 1 and Month 6 post vaccination. Post BCG vaccination, both circulating plasma levels and TB-antigen stimulated plasma levels of AMPs significantly increased at Month 1 and Month 6 compared to pre-vaccination levels in the elderly population. However, the association of AMP levels with latent TB (LTB) status did not exhibit statistical significance. Our findings indicate that BCG vaccination is linked to heightened circulating levels of AMPs in the elderly population, which are also TB-antigen-specific. This suggests a potential mechanism underlying the immune effects of BCG in enhancing host defense against TB.

Gene expression responses of CF airway epithelial cells exposed to elexacaftor/tezacaftor/ivacaftor (ETI) suggest benefits beyond improved CFTR channel function.

Gene expression responses by CF AEC exposed to ETI suggest that in addition to improving CFTR channel function, ETI is likely to increase resistance to bacterial infection by increasing levels of beta defensin 1 (hBD-1). ETI may also reduce lung damage by suppressing MMP10, and reduce airway inflammation by repressing proinflammatory cytokine secretion by AEC cells.

Human Beta Defensin-2 mRNA and Proteasome Subunit β Type 8 mRNA Analysis, Useful in Differentiating Skin Biopsies from Atopic Dermatitis and Psoriasis Vulgaris Patients.

(1): Atopic dermatitis and psoriasis vulgaris are chronic, inflammatory diseases. Clinical presentation usually leads to a proper diagnosis, but sometimes neither clinical examination nor histopathological evaluation can be conclusive. Therefore, we aimed to build up a novel diagnostic tool and check it for accuracy. The main objective of our work was to differentiate between healthy skin (C), atopic dermatitis (AD) and psoriasis vulgaris (PV) biopsies on the base of involucrin (IVL) and human β-defensin-2 (hBD-2) concentrations and their mRNA, as well as mRNA for TPP2 and PSMB8. (2): ELISA for IVL and hBD-2 proteins and Real-time PCR for the relative expression of mRNA for: IVL (IVL mRNA), hBD-2 (hBD-2 mRNA), PSMB8 (PSMB8 mRNA) and TPP2 (TPP2 mRNA), isolated from skin biopsies taken from AD and PV patients and healthy volunteers were performed. (3): hBD-2 mRNA and PSMB8 mRNA correlated with some parameters of clinical assessment of inflammatory disease severity. hBD-2 mRNA expression, exclusively, was sufficient to distinguish inflammatory skin biopsies from the healthy ones. (4): hBD-2 mRNA and PSMB8 mRNA analysis were the most valuable parameters in differentiating AD and PV biopsies.

Protective Effect of Coated Benzoic Acid on Intestinal Epithelium in Weaned Pigs upon Enterotoxigenic Escherichia coli Challenge.

The study was designed to investigate the protective effect of dietary supplementation with coated benzoic acid (CBA) on intestinal barrier function in weaned pigs challenged with enterotoxigenic Escherichia coli (ETEC). Thirty-two pigs were randomized to four treatments and given either a basal diet or a basal diet supplemented with 3.0 g/kg CBA, followed by oral administration of ETEC or culture medium. The results showed that CBA supplementation increased the average daily weight gain (ADWG) in the ETEC-challenged pigs (p < 0.05). CBA also increased the serum activity of total superoxide dismutase (T-SOD) and the total antioxidant capacity (T-AOC), as it decreased the serum concentrations of endotoxin, interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the ETEC-challenged pigs (p < 0.05). Interestingly, the CBA alleviated the ETEC-induced intestinal epithelial injury, as indicated by a reversal of the decrease in D-xylose absorption and a decrease in the serum levels of D-lactate and diamine oxidase (DAO) activity, as well as a decrease in the quantity of apoptotic cells in the jejunal epithelium following ETEC challenge (p < 0.05). Moreover, CBA supplementation significantly elevated the mucosal antioxidant capacity and increased the abundance of tight junction protein ZO-1 and the quantity of sIgA-positive cells in the jejunal epithelium (p < 0.05). Notably, CBA increased the expression levels of porcine beta defensin 2 (PBD2), PBD3, and nuclear factor erythroid-2 related factor 2 (Nrf-2), while downregulating the expression of toll-like receptor 4 (TLR4) in the jejunal mucosa (p < 0.05). Moreover, CBA decreased the expression levels of interleukin-1β (IL-1β), myeloid differentiation factor 88 (MyD88), and nuclear factor-kappa B (NF-κB) in the ileal mucosa upon ETEC challenge (p < 0.05). These results suggest that CBA may attenuate ETEC-induced damage to the intestinal epithelium, resulting in reduced inflammation, enhanced intestinal immunity and antioxidant capacity, and improved intestinal epithelial function.

Acute salivary antimicrobial peptide secretion response to different exercise intensities and durations.

Antimicrobial peptides, key players of innate mucosal immunity in the oral cavity, exert antibacterial and bacteriolytic effects. This study aimed to clarify the effects of acute exercise at different intensities and durations on salivary antimicrobial peptide levels. In a randomized crossover trial, 14 young healthy untrained men performed intensity trials [cycling at 35%, 55%, and 75% of maximal oxygen uptake (V̇o2max) for 30 min] and duration trials (cycling at 55% V̇o2max for 30, 60, and 90 min). Saliva samples were collected at baseline and 0 and 60 min after exercise. In intensity trials, the change in salivary lactoferrin levels from baseline to 0 min after 30 min exercise was greater at 75% V̇o2max exercise intensity compared with that at 35% V̇o2max. Furthermore, the change in salivary human β-defensin-2 (HBD-2) levels was greater at 75% V̇o2max compared with that at 35% and 55% V̇o2max. Salivary lysozyme levels increased after exercise, independent of exercise intensity. However, salivary LL-37 levels did not change after exercise at any intensity. In addition, in duration trials, the change in salivary levels of LL-37 and HBD-2 from baseline to 0 min after exercise at 55% V̇o2max was greater after 60 and 90 min of exercise compared with that after 30 min of exercise. However, salivary lactoferrin and lysozyme levels increased after exercise, independent of exercise duration. Our findings suggest that secretory responses to acute exercise with exercise intensity and duration differ among salivary antimicrobial peptides.NEW & NOTEWORTHY We investigated the effects of acute exercise at different intensities and durations on the immune response to salivary antimicrobial peptides in young healthy men. Levels of four salivary antimicrobial peptides increased after exercise dependently or independently of exercise intensity and duration, whereas some salivary antimicrobial peptides did not change after exercise. These findings suggest that the secretory responses to acute exercise with different intensities and durations differ among salivary antimicrobial peptides.

Alterations in the gut microbiota of Eimeria infected broiler chickens fed diets supplemented with varying levels of dietary calcium and phosphorus, along with 25-hydroxycholecalciferol

The purpose of the study was to investigate the effects of the reduced dietary calcium (Ca) and phosphorus (P) level and supplementation of 25-hydroxycholecalciferol (25-OHD3) on the expression of vitamin D receptor (VDR) and antimicrobial peptides and gut microbiota of broiler chickens with/without Eimeria challenge. A total of 576 fourteen-day-old broiler chicks were randomly allocated according to a 2 × 2 × 2 factorial design with main effects including Eimeria challenging (125,000 Eimeria acervulina, 25,000 Eimeria maxima, and 25,000 Eimeria tenella), dietary Ca and P levels (0.84% Ca and 0.42% available P or 0.64% Ca and 0.22% available P), and supplementation of 25-OHD3 (3,000 IU/kg) of 6 replicates. Three-way ANOVA was performed, and the effects of 3 main factors and their interactions were investigated. The reduced dietary Ca and P level downregulated cathelicidins 3 (CATH3) in the upper jejunum in the Eimeria challenging condition (interaction; P < 0.05). The reduced dietary Ca and P level decreased the relative mRNA expression of jejunal avian beta defensin 5 (AvBD5) in the Eimeria challenging condition (interaction; P < 0.05). The reduced dietary Ca and P level tended to decrease the relative mRNA expression of jejunal AvBD9 in the Eimeria challenging condition (interaction; P = 0.051). The reduced dietary Ca and P level decreased observed features (alpha diversity parameter for richness) in the upper jejunal microbiota in the Eimeria challenging condition (interaction; P < 0.05). The supplementation of 25-OHD3 decreased the relative abundance of the phylum Bacteroidetes (P < 0.05) and increased the relative abundance of the family Ruminococcaceae (P < 0.05) in the cecal digesta. The supplementation of 25-OHD3 decreased the serum endotoxin level in the Eimeria challenging condition (interaction; P < 0.05). Therefore, the reduced dietary Ca and P level modulated the upper jejunal microbiota via modulating the expression of antimicrobial peptides, and the supplementation of 25-OHD3 favorably modulated the cecal microbiota in broiler chickens with/without Eimeria challenge.