Expression Of Human Beta-defensin Research Articles

Correlation and Clinical Significance of HBD-2 and CXCL-1/2 Levels at Skin Lesions with Psoriasis Vulgaris Severity

ABSTRACT Objective This study was performed to explore the clinical significance of the expression of human beta-defensin 2 (HBD-2) and chemokine ligand 1/2 (CXCL-1/2) in psoriasis vulgaris. Methods This study retrospectively included the study group (n = 160) and control group (n = 100) for analysis. The levels of inflammatory indicators, blood biochemical indicators, and immune indicators using ELISA. The psoriasis area and severity index (PASI) was used to evaluate disease severity. Levels of HBD-2, CXCL-1, CXCL-2 and CCL20 were determined by RT-PCR. The correlations of HBD-2, CXCL-1 and CXCL-2 levels with CCL20 and PASI scores were analyzed. The diagnostic value of HBD-2, CXCL-1 and CXCL-2 in psoriasis vulgaris was analyzed by ROC curve. Results HBD-2, CXCL-1 and CXCL-2 were highly expressed in the lesions of psoriasis vulgaris patients, and were positively correlated with CCL20 and PASI score. HBD-2, CXCL-1 and CXCL-2 alone or in combination had high diagnostic value for psoriasis vulgaris and severe psoriasis, and the combined diagnostic value of the three was higher than that of a single indicator. Conclusion HBD-2, CXCL-1, and CXCL-2 levels are closely related to the severity of psoriasis vulgaris and can effectively diagnose the occurrence and progression of psoriasis vulgaris.

109 UV-induced expression of human beta-defensin 124 (DEFB124), a novel antimicrobial peptide with anti-S. aureus potential: is it a mediator of phototherapy in AD?

109 UV-induced expression of human beta-defensin 124 (DEFB124), a novel antimicrobial peptide with anti-S. aureus potential: is it a mediator of phototherapy in AD?

Differential expression of antimicrobial peptides in human fungal keratitis.

To analyze a series of antimicrobial peptides (AMPs) in corneal tissue from individuals with fungal keratitis (FK) during the active phase of the fungus infection and after healing. Patients undergone lamellar keratoplasty for the treatment of severe FK or corneal scar had their corneal buttons sampled. Quantitative real-time polymerase chain reaction (PCR) was used to ascertain the gene expression of human beta-defensin (HBD)-1, -2, -3, -9, S100A7, 8, 9, and LL-37. All AMPs' messenger ribonucleic acid (mRNA) expression was considerably elevated in all samples (n=12). In contrast to controls, where HBD-2, -3, and S100A7 mRNAs were expressed at very low levels, it was discovered that HBD-1, -9, S100A8, S100A9, and LL-37 were constitutively expressed in all healed samples (n=4). HBD-1, -2 -3, S100A7, and LL-37 mRNAs were significantly increased in all active FK samples (n=8). The levels of HBD-9, S100A8, and S100A9 mRNAs were moderately upregulated in all active FK samples. Subgroup comparison showed that HBD-2 was significantly increased in Fusarium keratitis samples (n=5), and LL-37 mRNAs were significantly enhanced in Aspergillus keratitis samples (n=3). Whereas there was not significantly increased of HBD-1, -3, -9, S100A7, 8, 9 mRNA in Aspergillus keratitis samples compared with Fusarium keratitis samples. AMPs expression is increased in active FK, but not all AMPs are equally expressed. HBD-2 and LL-37 expression levels are the highest, showing some specificity of AMP expression related to FK. Human AMPs, particularly HBD-2 may play a significant role in Fusarium keratitis and LL-37 might be the key player in Aspergillus keratitis.

Induction of Endogenous Antimicrobial Peptides to Prevent or Treat Oral Infection and Inflammation

Antibiotics are often used to treat oral infections. Unfortunately, excessive antibiotic use can adversely alter oral microbiomes and promote the development of antibiotic-resistant microorganisms, which can be difficult to treat. An alternate approach could be to induce the local transcription and expression of endogenous oral antimicrobial peptides (AMPs). To assess the feasibility and benefits of this approach, we conducted literature searches to identify (i) the AMPs expressed in the oral cavity; (ii) the methods used to induce endogenous AMP expression; and (iii) the roles that expressed AMPs may have in regulating oral inflammation, immunity, healing, and pain. Search results identified human neutrophil peptides (HNP), human beta defensins (HBD), and cathelicidin AMP (CAMP) gene product LL-37 as prominent AMPs expressed by oral cells and tissues. HNP, HBD, and LL-37 expression can be induced by micronutrients (trace elements, elements, and vitamins), nutrients, macronutrients (mono-, di-, and polysaccharides, amino acids, pyropeptides, proteins, and fatty acids), proinflammatory agonists, thyroid hormones, and exposure to ultraviolet (UV) irradiation, red light, or near infrared radiation (NIR). Localized AMP expression can help reduce infection, inflammation, and pain and help oral tissues heal. The use of a specific inducer depends upon the overall objective. Inducing the expression of AMPs through beneficial foods would be suitable for long-term health protection. Additionally, the specialized metabolites or concentrated extracts that are utilized as dosage forms would maintain the oral and intestinal microbiome composition and control oral and intestinal infections. Inducing AMP expression using irradiation methodologies would be applicable to a specific oral treatment area in addition to controlling local infections while regulating inflammatory and healing processes.

GCF levels of human beta defensins 1and 3 in diabetic and non diabetic patients with chronic periodontitis

Background: Human beta defensins (HβDs) expression in healthy tissues may aid in preventing the onset and/or progression of inflammatory disease.Type 2 diabetes mellitus (type 2 DM) and periodontal disease encompass inflammatory changes. HβDs in periodontitis patients is differentially regulated by periodontal pathogens, commensal organisms, host derived factors and inflammatory mediators. Additionally hyperglycaemia might influence this inflammatory state. Aim: To evaluate the role of the expression of human beta-defensin 1 and 3 levels in gingival crevicular fluid samples of gingivitis and chronic periodontitis patients with various glycaemic status. Methods and Material: In this study a total number of 140 patients were divided into five groups (Group I-V) consisted of 28 patients under each group. Group I consisted of 28 Type 2 Diabetes mellitus (DM) patients with well controlled glycemic status (> 6.5 <7%HbA1C) and chronic periodontitis(CP),Group II compromised of 28 type 2 DM-CP patients(7-8% HbA1C),Group III had 28 Type 2 DM-CP patients (>8%HbA1C).The concentrations of HβD-1 and HβD-3 were estimated using ELISA. Results: HβD-1 and 3 levels are higher in gingivitis subjects than periodontitis. Significantly higher levels of HβD-3 expression were found in the healthy tissues compared to the diseased ones.

Screening and characterization of active herbal extracts and components stimulating human beta defensing (hBD-3) expression from HaCaT cells

Various antimicrobial peptides play important roles as barrier function against bacteria and as innate host immunity. Several major studies have shown reduced expression of human beta defensin (hBD-3) in atopic dermatitis, highlighting the importance of hBD-3. In course of our search for hBD-3 stimulatory herbal souces and biologically active components, we screened 400 oriental herbal extracts using a range of real-time PCRs in the human keratinocyte cell line, HaCaT cells. We found that only six herbal extracts from Rubi Fructus, Lysimachiae Foenumgraeci Herba, Ailanthi Radicis Cortex, Galla Rhois, Peucedani Radix and Albizziae Cortex showed significant hBD-3 stimulatory effects. Although it did not have antimicrobial activity due to the low cytotoxic effect of Ailanthi Radicis Cortex, we further identified hBD-3-stimulating active ingredient in this extract. Efficient activity profiling based on HPLC was performed to identify active components from the extract of Ailanthi Radicis Cortex, and an hBD-3 stimulatory quassinoid component, ailanthone was identified by spectroscopic analysis. The present results show that Ailanthi Radicis Cortex and ailanthone have the potential to stimulate the expression of hBD-3 in HaCaT cells. These findings suggest new possibilities to control bacterial over-colonization found in atopic dermatitis using hBD-3 stimulatory herbal sources and ailanthone.

Opposing Roles of Antimicrobial Peptides in Skin Cancers.

Antimicrobial peptides (AMPs), also known as host defense peptides, are ubiquitous naturally occurring molecules secreted by various cell types of the body. In the skin, AMPs serve as a first-line innate immune defense against exogenous microorganisms, and they orchestrate adaptive immune responses to exert several immunomodulatory functions. Emerging evidence indicates that AMPs not only contribute to certain inflammatory skin diseases but also play a role in skin tumor carcinogenesis. Available data support the hypothesis that AMPs possess both pro-tumor and anti-neoplastic properties. Although inconsistent observations reported by multiple studies make it challenging to summarize the precise roles of AMPs in cancer, the differential expression of AMPs in skin cancers, such as the increased expression of human beta-defensins in squamous cell carcinoma and the ability of cathelicidin LL-37 to induce malignant melanoma cell invasion, implies they have procancer activities. On the other hand, the observation that certain AMPs show cytotoxic activity against cancer cells of the colon and kidney suggests their inherent antitumor properties. In this review, we describe the roles and mechanisms of AMPs in skin cancer development. We believe that further research is needed to elucidate the impact of these AMPs in skin cancer biology and to explore their potential roles as diagnostic/prognostic biomarkers and as novel therapeutic targets.

STEM-20. THE ROLE OF HUMAN BETA DEFENSINS IN THE CLONOGENICITY OF GLIOBLASTOMA MULTIFORME

Abstract INTRODUCTION Glioblastoma (GBM) is the most common primary malignant brain tumor with a median overall survival of 12-15months. GBM aggressiveness and poor response to treatment are often attributed to a small population of stem-like cells referred to as glioma stem cells (GSCs). Human Beta-Defensins (HBDs), a family of small molecules initially thought to function as anti-microbials, have been implicated in various cancers with functions that are cancer type specific, including proinflammatory and immunosuppressive roles. GOAL: This study aimed to elucidate HBDs expression in GSCs and differentiated gioma cells (DGSs). METHODS We identified HBD2 and HBD3 in glioma and GSCs and DGSs by immunofluorescence (IF) and immunohistochemistry (IHC). We also assessed the role HBD2/3 play in GBM oncogenesis by investigating their effects on the self-renewal capacity of GSCs. RESULTS HBD2 and HBD3 are found in both bulk tumor and GSCs by IHC and IF. Expression of HBD2 and HBD3 mRNA levels are elevated in GBM patient tissue in comparison to lower grade gliomas by 16.2 & 1.9 fold (respectively; p < 0.0001). Additionally, transcriptional expression of HBD2 and HBD3 are increased by 0.68 and 1.15 fold respectively in GSCs versus autologous DGCs (p < 0.05; p< 0.005 respectively), suggesting a role for HBD 2/3 in oncogenesis. Interestingly however, HBD2 and HBD3 pretreatment of GSC cell lines showed decreased self-renewal capacity by 34.2 and 66.4% (p < 0.001), determined by the reduced number of large neurospheres ( >250mm). CONCLUSIONS Our results demonstrate the presence of HBD2/3 in GBM samples by IHC and IF with increased HBD2/3 mRNA expression in GBM samples. Interestingly DGCs contain less HBD2/3 than their GCS counterparts, and clonogenicity assays demonstrate a decrease in oncogenesis, suggesting that HBD’s role in proliferation and clonogenicity of DGCs and GSCs may be context dependent, leading to additional questions and future studies.

Human beta-defensin-1 rs2738047 polymorphism is associated with shisha smoking risk among Saudi population

Human β-defensin (HBD), a member of the antimicrobial peptides, is essential for respiratory epithelial cells’ microbial defense, and is affected by cigarette smoking (CS). Its expression is upregulated by stimulation from microbes or inflammation. Genetic polymorphisms in the HBD-1 gene have been implicated in the development of various smoking-related diseases, including chronic obstructive pulmonary disease and asthma. Thus, we sought to analyze possible associations between HBD-1 single-nucleotide polymorphism (SNP) in HBD-1 gene and CS in ethnic Saudi Arabian subjects. Variants rs1047031 (C/T), rs1799946 (C/T), rs2738047 (C/T), and rs11362 (C/T) were investigated by genotyping 575 blood specimens from males and females, smokers/non-smokers: 288/287. The CT and CT+TT genotypes of rs1799946 presented an ~5-fold increased correlation with CS among the female smokers, compared with the female controls (OR = 5.473, P = 0.02003; and OR = 5.211, P = 0.02028, respectively), an observation similar to rs11362 SNP in female smokers, but with protective effects in TT genotype, compared with the CC reference allele (OR = 0.143, P = 0.04368). In shisha smokers, the heterozygous CT and the CT/TT genotype of rs2738047 polymorphism showed the same results with ~3-fold increased correlation with CS (OR = 2.788; P = 0.03448), compared with the cigarette smokers category. No significant association was shown in genotypic distributions and allelic frequencies of rs1047031. Further investigations, including large study samples, are required to investigate the effects of shisha on human beta-defensin expression and protein levels.

Dysregulation of human beta-defensin-3 expression in the peripheral blood of patients with sepsis.

Objectives:Sepsis is a serious medical condition caused by the body’s systemic inflammatory response to infections. The antimicrobial peptides, human beta-defensins, play a key role in modulating host immune responses, and aberrant expression of human beta-defensins has been implicated in many infections and inflammatory diseases. However, little is known about the expression of human beta-defensin-3 in systemic infectious diseases.Methods:We investigated the gene expression and protein level of human beta-defensin-3 in peripheral whole blood from 107 participants—67 patients with sepsis and 40 healthy controls—and evaluated the feasibility of human beta-defensin-3 as an indicator for sepsis. Total RNA was extracted from peripheral blood samples, and relative mRNA expression of human beta-defensin-3 was determined by reverse transcription-quantitative polymerase chain reaction. Plasma concentration of human beta-defensin-3 was measured by enzyme-linked immunosorbent assay. Pearson’s correlation analysis was performed to assess the relationship between human beta-defensin-3 mRNA and protein levels. Receiver operating characteristic analysis was performed to evaluate the value of human beta-defensin-3 as a biomarker for sepsis.Results:Human beta-defensin-3 mRNA expression was significantly downregulated in sepsis patients compared to controls (p = 0.001). The mean fold change of mRNA expression (±standard error) was 0.82 ± 0.63 in sepsis patients and 1.39 ± 1.09 in controls. Plasma concentration of human beta-defensin-3 (pg/mL) was significantly lower in sepsis patients compared to healthy controls (p = 0.039). The mean protein concentration (±standard error) was 539.6 ± 39.4 in sepsis patients and 715.5 ± 53 in controls. There was a significant correlation between human beta-defensin-3 mRNA expression and the corresponding protein level in sepsis patients (r = 0.358, p = 0.04), but not in healthy controls (r = 0.124, p = 0.51). For discriminating sepsis patients from healthy controls, the area under the receiver operating characteristic curve was 0.722 (95% confidence interval: 0.597–0.847, p = 0.002) for human beta-defensin-3 mRNA and 0.689 (95% confidence interval: 0.557–0.827, p = 0.009) for human beta-defensin-3 protein.Conclusion:This is the first study to show the downregulation of human beta-defensin-3 gene expression and protein level in sepsis, which may contribute to the complex immunological imbalance in sepsis. The significant correlation between human beta-defensin-3 mRNA expression and protein concentration suggests that mRNA expression could be used to predict protein level. Our study also showed a potential role of human beta-defensin-3 as a blood-based biomarker for sepsis. More studies on the clinical significance of human beta-defensin-3 in sepsis could further support a biomarker development.

Antimicrobial peptides in human corneal tissue of patients with fungal keratitis

BackgroundFungal keratitis (FK) is the leading cause of unilateral blindness in the developing world. Antimicrobial peptides (AMPs) have been shown to play an important role on human ocular surface (OS)...

Salivary human beta-defensins and cathelicidin levels in relation to periodontitis and type 2 diabetes mellitus

Objective: Type 2 diabetes mellitus (T2DM) is a well-defined risk factor of periodontitis and it can affect expression of human beta-defensins (hBDs) and cathelicidin (LL-37) as well. The aim of the present study was to evaluate the impact of periodontitis and T2DM on salivary concentrations of these antimicrobial peptides.Material and methods: Unstimulated saliva samples, together with full-mouth periodontal recordings were collected from 92 individuals with periodontitis (63 with T2DM and 21 smokers) and 86 periodontally healthy controls (58 with T2DM and 21 smokers). Salivary hBD-1, -2, -3, LL-37, and advanced glycalization end products (AGE) concentrations were measured by enzyme-linked immunosorbent assay.Results: Among the periodontitis patients, T2DM group demonstrated lower levels of hBD-1 (p = .006), hBD-2 (p < .001) and hBD-3 (p < .001), and higher levels of LL-37 (p < .001) compared to systemically healthy controls. When only periodontally healthy controls were included into the analysis, higher hBD-1 (p = .002) and LL-37 (p < .001) levels were found in T2DM patients in comparison to systemically healthy controls. Salivary LL-37 levels were associated with HbA1c and periodontitis, while hBD-2, hBD-3 and levels associated only with HbA1c.Conclusion: In the limits of this study, hyperglycaemia can be proposed as a regulator of salivary hBD and cathelicidin levels. Periodontitis, on the other hand, affects only salivary LL-37 levels.

The guardians of the periodontium—sequential and differential expression of antimicrobial peptides during gingival inflammation. Results from in vivo and in vitro studies

To evaluate the sequential and differential expression of a variety of antimicrobial peptides (AMPs) during the development of an experimentally induced gingivitis in humans. In twenty healthy volunteers, gingival inflammation was induced by abstention from oral hygiene at 6 teeth. Bleeding on probing (BOP) and plaque index (PI) were assessed, and gingival biopsies and gingival crevicular fluid (GCF) were collected at 8 different time points (t0-t35). Gingival epithelial cells (GECs) were stimulated with various receptor agonists. In biopsies and GECs, mRNA expression of human beta-defensins (hBD-2, hBD-3), CC-chemokine ligand 20 (CCL20), S100A7/psoriasin (S100A7), and calgranulin A/B (S100A8, S100A9) was evaluated using real-time PCR, and protein profiles were measured by ELISA. Statistical analysis was performed using non-parametric tests. The clinical parameters BOP, PI and GCF increased over time (p<0.0001). Tissue AMP mRNA expression was elevated, but at different and AMP-specific time points (p<0.05). Protein analysis revealed a similar expression pattern for hBD-2 and CCL20 in GCF (p<0.05). In GECs, multiple receptor stimulation was required to induce AMP gene expression (p<0.0001). For the first time, this study showed the sequential and differential expression of AMPs during a developing inflammation in vivo providing further evidence for their role as guardians of a healthy periodontium.

Evaluation of the effect of pulsed dye laser on chronic psoriatic plaque

Background: Psoriasis is one of the most prevalent immune mediated skin diseases. Selective vascular destruction by pulsed dye laser is considered one of effective laser treatments. Objective: Evaluation of the therapeutic effect of pulsed dye laser in treatment of chronic psoriatic plaque lesions and changes in human beta defensin-2(HBD-2) expression in the psoriatic patients before and after treatment, with correlation to the clinical improvement.Methods: Twenty patients with psoriatic plaque on right side treated by laser and another plaque on left side similar in size and its severity score served as control receiving no treatment. Each patient received 3–4 sessions of pulsed dye laser 595 nm. Follow up was done for three months.Results: There was statistically significant difference between the two psoriatic plaques (untreated and treated; A and B; p = .004). There was significant decrease in HBD-2 expression after treatment. No recurrence was observed during follow up period.Conclusion: The pulsed dye laser is safe, effective, and tolerable treatment for resistant stable localized plaque psoriasis with minimal side effects and prolonged recurrence period.

Dynamic changes of Th1/Th2/Th17 cytokines and human beta defensin 2 in HIV-infected patients with oral candidiasis during the first year of highly active anti-retroviral therapy

ObjectiveTo investigate the change of Th1/Th2/Th17 cytokines and human beta defensin 2 (HBD-2) in HIV-infected patients with oral candidiasis (OC) and gather information about OC-specific immunity. DesignDuring the 1st year of highly active anti-retroviral therapy (HAART), 25 HIV-infected patients were followed up at the baseline, 3rd, 6th, 12th month. At each visit, oral manifestations were examined; oral rinses were collected and cultured for Candida; peripheral venous blood was taken to determine CD4 + T cell counts and HIV RNA viral load (VL); both unstimulated whole saliva and peripheral venous blood were taken to determine cytokine (IL-4, IL-17(A/F), IFN-γ) and HBD-2 levels. Twenty-five healthy individuals were enrolled as control. ResultsHIV-infected patients displayed lower levels of IL-17(A/F) and IFN-γ but higher level of IL-4 and HBD-2 compared with healthy controls. During the 1st year of HAART, salivary IL-17(A/F) and IFN-γ were in uptrend, whereas salivary IL-4 and salivary HBD-2 were in downtrend. Serum cytokines all show no significant changes. After 1 year of HAART, serum, salivary IL-4 level and salivary IL-17(A/F) showed no significant difference from healthy controls. HIV-infected patients with OC had a higher IL-4 level but lower IFN-γ and IL-17(A/F) levels than those without OC since the 3rd month of HAART. The occurrence of OC was negatively correlated to IL-17(A/F) and IFN-γ, but positively correlated to IL-4. Salivary HBD-2 expression was up-regulated in HIV and might associate with Candida albicans. ConclusionsIn HIV-infected patients, the decrease of IL-17 and IFN-γ, and the increase of IL-4 in local and systemic level could influence the prevalence of OC. Salivary HBD-2 may also play an important role against OC.

Early type I Interferon response induces upregulation of human β-defensin 1 during acute HIV-1 infection.

HIV-1 is able to evade innate antiviral responses during acute infection to establish a chronic systemic infection which, in the absence of antiretroviral therapy (ART), typically progresses to severe immunodeficiency. Understanding these early innate immune responses against HIV-1 and their mechanisms of failure is relevant to the development of interventions to better prevent HIV-1 transmission. Human beta defensins (HBDs) are antibacterial peptides but have recently also been associated with control of viral replication. HBD1 and 2 are expressed in PBMCs as well as intestinal tissue, but their expression in vivo during HIV-1 infection has not been characterized. We demonstrate that during acute HIV-1 infection, HBD1 but not HBD2 is highly upregulated in circulating monocytes but returns to baseline levels during chronic infection. HBD1 expression in monocytes can be induced by HIV-1 in vitro, although direct infection may not entirely account for the increase in HBD1 during acute infection. We provide evidence that HIV-1 triggers antiviral IFN-α responses, which act as a potent inducer of HBD1. Our results show the first characterization of induction of an HBD during acute and chronic viral infection in humans. HBD1 has been reported to have low activity against HIV-1 compared to other defensins, suggesting that in vivo induced defensins may not significantly contribute to the robust early antiviral response against HIV-1. These data provide important insight into the in vivo kinetics of HBD expression, the mechanism of HBD1 induction by HIV-1, and the role of HBDs in the early innate response to HIV-1 during acute infection.

Human Beta Defensins 1, 2 and 3 Produced by Amniochorion Membranes Is Similar in Term and Preterm Delivery

Amniochorion membranes were collected from 25 pregnant women at preterm labor, in the presence or not of Preterm Premature Rupture of Membranes (PPROM) and 27 pregnant women at term in the presence at labor, in order to quantify the expression and to evaluate the immunoreactivity of human beta defensins (HBD)1, HBD2, HBD3 and HBD4 in amniochorion membranes from pregnancies complicated by spontaneous prematurity. The HBDs were evaluated by immunohistochemistry, real time quantitative PCR and ELISA. Statistical analyses were performed using Chi-squared and Mann Whitney tests. There was no significant difference in HBDs expression between study and control groups: HBD1 (Md = 0.62 (0.0 - 105.0) vs Md = 0.80 (0.02 - 25.0); p = 0.85), HBD2 (Md = 0.17 (0.0 - 5.2) vs Md = 0.0 (0.0 - 43.2); p = 0.16), HBD3 (Md = 0.11 (0.0 - 140.5) vs Md = 0.06 (0.0 - 972.1); p = 0.91). Also, HBD1, HBD2 and HBD3 protein expression was not significant different between the groups: HBD1 (1.32 pg/mL (0.0 - 1.85) vs 1.08 pg/mL (0.04 - 2.22); p = 0.67), HBD2 (0.00 pg/mL (0.0 - 1.74) vs 0.02 pg/mL (0.0 - 1.24); p = 0.69), HBD3 (0.04 pg/mL (0.0 - 1.05) vs 0.09 pg/mL (0.0 - 1.05); p = 0.63). The immunoreactivity of HBD1, HBD2 and HBD3 was observed in amnion, chorion and decidua cells from preterm and term pregnancies. Amniochorion membranes are sources of HBD1, HBD2 and HBD3 and their expressions are similar in term and preterm pregnancies.

Immune response in human chromoblastomycosis and eumycetoma - focusing on human interleukin-17A, interferon-gamma, tumour necrosis factor-alpha, interleukin-1 beta and human beta-defensin-2.

Knowledge regarding host immune response to chromoblastomycosis and eumycetoma is limited, particularly concerning cytokines and antimicrobial peptides production. This was a retrospective study of 12 paraffin-embedded tissue samples from patients diagnosed with chromoblastomycosis or eumycetoma from histological findings and tissue culture. DNA extraction and polymerase chain reaction (PCR) from tissues were done to evaluate human interleukin-17A (IL-17A), interferon-gamma (IFN-γ), tumour necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β) and human beta-defensin-2 (HBD-2) expressions. Human beta-actin primer was used for confirming DNA detection, and DNA extracted from psoriasis lesional skin samples was used as positive controls. The twelve paraffin-embedded sections used in this study consisted of five chromoblastomycosis and seven eumycetoma tissues. All PCR reactions showed beta-actin band at 51 bp in all clinical specimens, confirming adequate DNA levels in each reaction. As positive control, the psoriasis skin samples revealed bands for IL-17A at 174 bp, IFN-γ at 273 bp, TNF-α at 360 bp, IL-1β at 276 bp and HBD-2 at 255 bp. For the chromoblastomycosis and eumycetoma tissues, PCR analyses showed IL-17A band at 174 bp in two eumycetoma tissues and HBD-2 band at 255 bp in a chromoblastomycosis tissue. This study demonstrated IL-17A expression in human eumycetoma and HBD-2 expression in human chromoblastomycosis for the first time. However, their role in immune response remains to be elucidated.

Trefoil factor 3 (TFF3) from human breast milk activates PAR-2 receptors, of the intestinal epithelial cells HT-29, regulating cytokines and defensins.

Trefoil factors are effector molecules in gastrointestinal tract physiology. Each one improves healing of the gastrointestinal tract. Trefoil factors may be grouped into three classes: the gastric peptides (TFF1), spasmolytic peptide (TFF2) and intestinal trefoil factor (TFF3). Significant amounts of TFF3 are present in human breast milk. Previously, we have reported that trefoil factor 3 isolated from human breast milk produces down regulation of cytokinesand promotes human beta defensins expression in intestinal epithelial cells. This study aimed to determine the molecular mechanism involved. Here we showed that the presence of TFF3 strongly correlated with protease activated receptors 2 (PAR-2) activation in human intestinal cells. Intracellular calcium ((Ca2+)i)mobilization was induced by the treatment with: 1) TFF3, 2) synthetic PAR-2 agonist peptide. The co-treatment with a synthetic PAR-2 antagonist peptide and TFF3 eliminates the latter's effect. Additionally, we demonstrated the existence of interactions amongTFF3 and PAR-2 receptors through far Western blot and co-precipitation. Finally, down regulationof PAR-2 by siRNAresulted in a decrease of TFF3 induced intracellular (Ca2+)i mobilization, cytokine regulation and defensinsexpression. These findings suggest that TFF3 activates intestinal cells through PAR-2 (Fig. 4, Ref. 19).

Effects of whole cigarette smoke on human beta defensins expression and secretion by oral mucosal epithelial cells.

BackgroundCigarette smoke a recognized risk factor for many systemic diseases and also oral diseases. Human beta defensins (HBDs), a group of important antimicrobial peptides expressed by the epithelium, are crucial for local defense and tissue homeostasis of oral cavity. The aim of this study was to evaluate potential effects of whole cigarette smoke (WCS) exposure on the expression and secretion of HBDs by oral mucosal epithelial cells.MethodsImmortalized human oral mucosal epithelial (Leuk-1) cells were exposed to WCS for various time periods. HBD-1, -2 and -3 expression and subcellular localization were detected by real time qPCR, immunofluorescence assay and confocal microscopy. According to the relative fluorescent intensity, the expression levels of HBD-1, -2 and -3 were evaluated by digital image analysis system. The alteration of HBD-1, -2 and -3 secretion levels was measured by the Enzyme-Linked Immunosorbent Assay.ResultsWCS exposure remarkably attenuated HBD-1 expression and secretion while clearly enhanced HBD-2, -3 expression levels and HBD-2 secretion by Leuk-l cells. It appeared that there was no significant effect of WCS exposure on HBD-3 secretion.ConclusionsWCS exposure could modulate expression and secretion of HBDs by oral mucosal epithelial cells, establishing a link between cigarette smoke and abnormal levels of antimicrobial peptides. The present results may give a new perspective to investigate smoking-related local defense suppression and oral disease occurrence.