Inhibition Of IgE Production Research Articles

B cell intrinsic effects of the vitamin D receptor on IgE responses. (IRC10P.417)

Abstract B cells express the vitamin D receptor (VDR). VDR KO mice had higher levels of circulating total IgE than WT mice by 12 wks of age. Mice were sensitized using two OVA/alum injections. VDR KO mice mounted a significantly higher OVA-specific IgE response compared to WT mice. The data also showed that vitamin D deficient mice had higher OVA-specific IgE responses than vitamin D sufficient mice. IL-4, IL-5, IL-2, IFN-γ and IL-17 were not different in OVA re-stimulated spleen and mesenteric lymph nodes (MLN) in WT and VDR KO mice. However, VDR KO spleen and MLN produced significantly less IL-10 than WT. Furthermore B cells and not T cells were the major source of IL-10 in both the WT and VDR KO cultures. Experiments were done using T cell specific-VDR deletion (T-VDR KO), or B cell specific-VDR deletion (B-VDR KO). The serum IgE levels in T-VDR KO mice were comparable to that of WT mice. Conversely, B-VDR KO mice had higher total serum IgE levels than WT mice and, when immunized, produced significantly more OVA-IgE. In our current model, we propose that B cell-derived IL-10 is regulated by the VDR and that autocrine IL-10 is higher in WT mice, therefore inhibiting IgE production to a greater extent in WT than in VDR KO mice. The major finding of this work is that vitamin D receptor expression selectively regulates IgE levels in the serum via a direct effect of the VDR in B cells.

2-Methyl-1, 3, 6-Trihydroxy-9, 10-Anthraquinone Isolated from Rubia Cordifolia L Inhibits IgE Production

2-Methyl-1, 3, 6-Trihydroxy-9, 10-Anthraquinone Isolated from Rubia Cordifolia L Inhibits IgE Production

The beneficial properties of marine polysaccharides in alleviation of allergic responses.

Marine polysaccharides have been found as the principle component in cell wall structures of seaweeds or exoskeletons of crustaceans. Due to numerous pharmaceutical properties of marine polysaccharides such as antioxidant, anti-inflammatory, antiallergic, antitumor, antiobesity, antidiabetes, anticoagulant, antiviral, immunomodulatory, cardioprotective, and antihepatopathy activities, they have been applied in many fields of biomaterials, food, cosmetic, and pharmacology. Recently, several marine polysaccharides such alginate, porphyran, fucoidan, and chitin and its derivatives have been evidenced as downregulators of allergic responses due to enhancement of innate immune system, alteration of Th1/Th2 balance forward to Th1 cells, inhibition of IgE production, and suppression of mast cell degranulation. This contribution, therefore, focuses on antiallergic properties of marine polysaccharides and emphasizes their potential application as bioactive food ingredients as well as nutraceuticals for prevention of allergic disorders.

Regulation of IgE production and airway reactivity by CD4−CD8− regulatory T cells

The mechanisms of tolerance induction occurring in the course of allergen-specific immunotherapy have not been elucidated in full detail. Our study aimed to characterize high zone tolerance in mouse models of type I allergy and of allergic airway inflammation induced by subcutaneous sensitization of mice with high doses of the model allergen ovalbumin (OVA) without the use of adjuvant. Mice were immunized by subcutaneous injection of high doses (HD) of OVA or, for comparison, low doses (LD) of OVA in saline. HD-mice showed lower specific IgE, but augmented IgG in sera than LD-mice. Pre-treatment of mice with HD-OVA antigen-specifically inhibited IgE production subsequently induced by LD-OVA. OVA-restimulated splenocytes from HD-mice revealed hypoproliferation and impaired production of Th2-associated cytokines. HD-mice exhibited lower airway reactivity, goblet cell hyperplasia and mucus production, as well as IL-5 and IL-13 production in the lungs than LD-mice following local provocation. Recruitment of inflammatory cells into the airways was comparable, while the number of eosinophils in the bronchoalveolar lavage was substantially higher in HD-mice. Adoptive transfer of dnTC from HD-mice into naïve mice, which were subsequently sensitized with LD-OVA, suppressed IgE production in the recipients. The number of dnTC was higher in the spleens of HD-mice than LD-mice. In conclusion, our study demonstrates that subcutaneous sensitization of mice with high doses of allergen in the absence of adjuvant results in attenuated airway reactivity as compared with LD-sensitization and induces CD4−CD8− dnTC with regulatory function on IgE production.

Berberine and limonin suppress IgE production by human B cells and peripheral blood mononuclear cells from food-allergic patients

BackgroundCurrently, there is no satisfactory treatment for IgE-mediated food allergy. Food Allergy Herbal Formula 2 (FAHF-2) and butanol-purified FAHF-2 (B-FAHF-2) have been shown to protect against peanut-induced anaphylaxis and inhibit IgE synthesis in a murine model. ObjectiveTo determine which herbs and compounds in FAHF-2 and B-FAHF-2 suppress IgE production. MethodsThe effect of FAHF-2 and B-FAHF-2 on IgE production was determined using a human B-cell line (U266). Individual compounds were isolated and identified using column chromatography, liquid chromatographic mass spectrometry, and nuclear magnetic resonance techniques. The potency of compounds on IgE suppression were investigated using U266 cells and verified using human peripheral blood mononuclear cells (n = 25) from peanut-allergic patients. Epsilon germline transcript expression was determined. Phosphorylated IκBα level was analyzed using the In-Cell Western assay. The mRNA expression of signal transducer and activator of transcription-3, T-box transcription factor TBX21, interferon-γ, forkhead box P3, GATA-binding protein 3, interleukin-10, and interleukin-5 also were analyzed using real-time polymerase chain reaction. ResultsFAHF-2 and B-FAHF-2 inhibited IgE production by U266 cells. B-FAHF-2 was 9 times more effective than FAHF-2. Two compounds that inhibited IgE production were isolated from Philodendron chinensis and identified as berberine and limonin. Berberine was more potent and inhibited IgE production by peripheral blood mononuclear cells by 80% at 0.62 μg/mL. Berberine significantly inhibited ε-germline transcript expression by peripheral blood mononuclear cells. Phosphorylated IκBα level was significantly suppressed and mRNA expressions of T-box transcription factor TBX21 and signal transducer and activator of transcription-3 were significantly increased by berberine. ConclusionBerberine and limonin mediated IgE suppression. The mechanism by which berberine modulates ε-germline transcript expression might be through regulating the phosphorylated IκBα level and the expressions of signal transducer and activator of transcription-3 and T-box transcription factor TBX21. Trial RegistrationClinicaltrials.gov identifier NCT00602160.

25-hydroxvitamin D3 promotes the long-term effect of specific immunotherapy in a murine allergy model.

Calcitriol (1α,25-dihydroxyvitamin D3) is the active vitamin D metabolite and mediates immunological functions, which are relevant in allergy. Its therapeutic use is limited by hypercalcaemic toxicity. We have previously shown that the activation of the vitamin D receptor inhibits IgE production and that B cells can synthesize calcitriol from its precursor 25-hydroxyvitamin D3 (inactive precursor) [25(OH)D] upon antigenic stimulation. In this study, we address the impact of 25(OH)D on the development of type I sensitization and determine its role in allergen-specific immunotherapy. BALB/c mice were sensitized to OVA, under 25(OH)D-deficient or sufficient conditions. The humoral immune response over time was measured by ELISA. OVA-specific immunotherapy was established and studied in a murine model of allergic airway inflammation using lung histology, pulmonary cytokine expression analysis, and functional parameters in isolated and perfused mouse lungs. In 25(OH)D-deficient mice, OVA-specific IgE and IgG1 serum concentrations were increased compared with control mice. OVA-specific immunotherapy reduced the humoral immune reaction after OVA recall dose-dependently. Coadministration of 25(OH)D in the context of OVA-specific immunotherapy reduced the allergic airway inflammation and responsiveness upon OVA challenge. These findings were paralleled by reduced Th2 cytokine expression in the lungs. In conclusion, 25(OH)D deficiency promotes the development of type I sensitization and correction of its serum concentrations enhances the benefit of specific immunotherapy.

The Role of Hypoxia-Inducible Factor 1α in Allergic Rhinitis

Hypoxia-inducible factor (HIF) 1alpha is expressed under hypoxic conditions and plays an important role in immune and inflammatory responses. The role of HIF-1alpha in allergic airways has been investigated mainly in bronchial asthma. This study investigated the role of HIF-1alpha in mouse models and patients with allergic rhinitis (AR). Balb/c mice were sensitized with ovalbumin (OVA) and alum and were challenged intranasally with OVA. The HIF-1alpha inhibitor, 2-methoxyestradiol (2-ME), was administered intraperitoneally and multiple parameters of allergic responses were evaluated. HIF-1alpha and vascular endothelial growth factor (VEGF) mRNA and protein expression were also evaluated in patients with AR and a correlation analysis between mRNA expression and allergic symptoms was performed. In mouse models, the HIF-1alpha inhibitor 2-ME reduced allergic symptoms and eosinophilic infiltration into the nasal mucosa. 2-ME was found to suppress IgE production and inhibit local Th2 cytokine transcription in the nasal mucosa and systemic Th2 cytokine production by splenocytes. 2-ME also decreased HIF-1 and VEGF expression in nasal mucosa. An increase in HIF-1alpha and VEGF expression in the nasal mucosa of patients with AR was also observed. Our data suggest that HIF-1alpha plays an important role in mouse models and patients with AR. HIF-1alpha inhibitors induce antiallergic effects by decreasing both local and systemic Th2 cytokine (IL-4 and IL-5) production, IgE production, and eosinophil infiltration into the nasal mucosa in an AR model. HIF-1alpha and VEGF expression increased in the nasal mucosa of patients with AR, showing the role of HIF-1alpha in disease pathogenesis.

Chapter One - Marine-Derived Polysaccharides for Regulation of Allergic Responses

Polysaccharides are macromolecules made up of many monosaccharides joined together by glycosidic bonds. Polysaccharides from marine sources are widely distributed as the principle component in cell wall structures of seaweeds or exoskeletons of crustaceans. So far, marine polysaccharides have been used in many fields of biomaterials, food, cosmetic, and pharmacology. Especially, numerous pharmaceutical properties of marine polysaccharides have been revealed such as antioxidant, anti-inflammatory, antiallergic, antitumor, antiobesity, antidiabetes, anticoagulant, antiviral, immunomodulatory, cardioprotective, antihepatopathy, antiuropathy, and antirenalpathy activities. Recently, several marine polysaccharides such alginate, porphyran, fucoidan, and chitin and its derivatives have been found as modulators of allergic responses due to enhancing innate immune system, altering Th1/Th2 balance, inhibiting IgE production, and suppressing mast cell degranulation. This contribution, therefore, focuses specially on the immunomodulatory effect of marine polysaccharides and emphasizes their potential application as candidates of pharmaceuticals as well as nutraceuticals to prevent allergic disorders.

Poster 1013: IL-4R alpha antibody inhibits IgE production and airway remodeling in mouse model of house dust mite-induced eosinophilic asthma

Background Allergic asthma is associated with T helper 2 (Th2) immune response to an allergen. IL-4 and IL-13, key cytokines involved in Th2 immunity, promote eosinophilic inflammation and B cell isotype switching to IgE. House Dust Mite (HDM) allergen has been shown to induce allergic asthma in a mouse model via Th2 immune response that drives eosinophilia and influx of effector T cells into lung. An IL-4Raplha antibody that blocks signaling of IL-4 and IL-13 through both Type I (IL-4Ralpha/ IL-2Rgamma) and Type II (IL-4Ralpha/IL-13R) receptors, and an IL-13Ra2-Fc fusion protein that prevents binding and signaling of IL-13 via Type II receptor, were both tested in the HDM-induced mouse model of asthma. Methods Mice were sensitized daily for 10 days with HDM, rested for 2 weeks, followed by challenge with HDM three times weekly for 8 weeks. Both anti-mouse IL-4Ralpha; antibody (Ab) and fully human anti-human IL-4Raplha Ab (dupilumab) were tested in wild type mice and mice humanized for IL-4 and IL-4Ralpha, respectively. Mice were treated with 50 mg/kg of either Ab twice weekly, starting at week 7 of the experiment. A separate group received mouse IL-13Raplha2-Fc protein using the same dose and regimen. At the end of the experiment, cells obtained by left lung lobe tissue digest were analyzed by flow cytometry. Lung collagen indicative of an airway remodeling was quantified using a Sircol Dye assay, and cardiac lung lobe was used for microarray analysis of gene signature. Levels of HDM-specific IgG1 and total IgE in serum were analyzed by ELISA. Results HDM sensitization and challenge resulted in increased levels of IgE and HDM-specific IgG1. IgE increase was blocked by IL-4Ra Ab but not by IL-13Ralpha2-Fc treatment; HDM-specific IgG1 levels were not affected by either treatment. Collagen content in lung of mice treated with IL-4Ralpha mAb was similar to that observed in mock sensitized and challenged mice. Dupilumab treatment also prevented the influx of eosinophils and inflammatory dendritic cells into lung. HDM-induced changes in gene expression were largely normalized upon dupilumab treatment, as compared to mock treated and sensitized mice.

Midazolam Inhibits IgE Production through Suppression of Class Switch Recombination

Midazolam（MDZ）is reported to have immunomodulatory properties that affect immune cells. However, little is known about the effects of MDZ treatment on Immunoglobulin（Ig）E responses. Therefore, we examined whether MDZ is able to suppress total IgE production, followed by IgE class switch recombination（CSR）, in a mouse model. To assess the effects of MDZ on IgE-CSR, splenic B cells were cultured with LPS, IL-4 and anti-CD40 antibody in the presence or absence of MDZ for 72 h. Total-IgE, interferon-gamma（IFN-γ）responses and surface IgE-positive（sIgE+）B cells were analyzed by ELISA and flow cytometry. To confirm IgE-CSR, total RNA was isolated from splenic B cells and levels of CSR-associated molecules, such as germ-line transcript ε（εGLT）, germ-circle transcript ε（εCT）, activation-induced cytidine deaminase（AID）and inhibitor of differentiation 2（Id2） were compared. MDZ significantly decreased total IgE production and numbers of sIgE+ B cells. Significantly reduced levels of both εGLT- and εCT-specific mRNA were detected in MDZ-treated B cells. In contrast, Id2-specific mRNA transcript, which is a negative regulator for Ig-CSR, was increased on MDZ-treated B cells. Furthermore, MDZ-treated B cell significantly increased IFN-γ production and IFN-γRα expression. These results suggest that MDZ inhibits εGLT and εCT expression, and IgE synthesis via induction of IFN-γ production. MDZ may be useful for preventing IgE-mediated allergic diseases.

Listeria infection inhibits IgE production in regional lymph nodes by suppressing chemotaxis of basophils to lymph nodes

Infection with Listeria induces a dominant shift to the Th1 immune response and inhibits the Th2 response. Papain is frequently utilized in animal models of allergies. Papain administration induces chemotaxis of basophils to regional lymph nodes (LNs) and production of interleukin (IL)-4 by basophils, resulting in a Th2-dominant status and increased IgE production in LNs. In this model, production of immunoglobulin (Ig) E by LN cells is primarily controlled by IL-4 produced by basophils. Based on this model, it was postulated that Listeria monocytogenes (Lm) infection suppresses IgE production by LN cells. Therefore, the effects of Lm infection on a papain-induced mouse model of allergies were investigated. Following s.c. injection of papain, basophils transiently migrated to draining LNs because of the effects of chemokine (C-C) motif ligand (CCL) 24 and secreted IL-4, inducing a Th2 response. Lm infection blocked recruitment of basophils into the popliteal LNs by inhibiting CCL24 production. Papain-induced class switch recombination (CSR) to IgE is inhibited by Lm infection, whereas CSR to IgG1 is not affected by the same treatment. Therefore, the CSR of IgG1 to IgE is basophil-dependent, whereas the CSR of IgM to IgG1 is basophil-independent. Hence, Lm infection suppresses CSR to IgE without affecting CSR to IgG1.

Modulation of experimental atopic dermatitis by topical application of Gami-Cheongyeul-Sodok-Eum

BackgroundGami-Cheongyeul-Sodok-Eum (GCSE), an herbal formula of traditional Korean medicine, comprises nine herb components. GCSE has various biological activities such as anti-inflammatory, anti-bacterial and anti-viral activities. However, it is still unclear whether GCSE has any immunomodulatory effect on atopic dermatitis (AD).MethodsGCSE was treated to primary B cells and CD4+ T cells isolated from atopic mice to compare its inhibitory effects on IgE secretion and cytokine expression. Experimental AD was established by alternative treatment of 2, 4-dinitrochlorobenzene (DNCB) and house dust mite extract to the ears of BALB/c mice. GCSE was topically applied to ears of atopic mice every day for 3 weeks. AD progression was analyzed by measuring ear thickness, serum IgE level, histological examination of ear tissue by H&E staining and cytokine profile of CD4+ T cells and CD19+ B cells by real time PCR and ELISA.ResultsTreatment of GCSE significantly reduced IgE production and expression of AD associated pathogenic cytokines such as IL-4, IL-5, IL-10, IL-13, IL-17, TNF-α, and IFN-γ by lymphocytes isolated from AD-induced mice. Topical application of GCSE on the ears of AD-induced mice significantly reduced ear thickness, clinical score and lymphocytes infiltration to ears as compared to control group. GCSE treatment also reduced serum IgE level and the levels of major pathogenic cytokines such as IL-4, IL-5, IL-10, IL-13 and IL-17. In addition, GCSE treatment significantly increased Foxp3 expression level.ConclusionsThe protective effect of GCSE in experimental AD is mediated by inhibition of IgE production, by reduction in the levels of pathogenic cytokines and by induction of Foxp3, all of which are suggesting the beneficial effect of GCSE on modulating atopic dermatitis.

MINOCYCLINE BLOCKS ALLERGEN‐INDUCED EOSINOPHILIA AND PRODUCTION OF TH2 CYTOKINES AND IGE BY INTERFERING WITH THE T CELL RECEPTOR‐NF‐kB‐GATA‐3‐INTERLEUKIN (IL)‐4 AXIS IN A MURINE ASTHMA MODEL WITHOUT AN EFFECT ON PARP

Minocycline protects against asthma independently of its antibiotic function. Minocycline was recently reported as a potent PARP inhibitor. In an animal model of asthma, a single administration of minocycline conferred excellent protection against ovalbumin (OVA)‐induced airway eosinophilia, production of mucus, Th2 cytokine and OVA‐specific IgE. These effects were unrelated to a potent inhibition of PARP; the drug moderately reduced PARP activation by modulating oxidative stress‐associated DNA damage. Minocycline failed to efficiently block direct PARP activation in lungs of MNNG‐treated mice or H2O2‐treated cells. Minocycline blocked OVA‐specific IgE production in B cells potentially by modulating T cell‐receptor (TCR)‐linked production at IL‐4 mRNA level but not through a modulation of the IL‐4‐JAK‐STAT‐6 axis, IL‐2 production, or NFAT‐1 activation. Such IL‐4 production blockade was linked to a preferential inhibition of the NF‐κB activation arm of TCR but not GSK3, Src, p38 MAPK, or ERK1/2. Inhibition of NF‐κB activation was linked to a complete blockade of TCR‐stimulated GATA‐3 expression, a pivotal transcription factors for IL‐4 expression. These results show that minocycline inhibits IgE production and asthma traits by modulating TCR function particularly by inhibiting the signaling pathway leading to NF‐κB activation, GATA‐3 expression, and subsequent IL‐4 production.

Suppressive Effect of Wild Saccharomyces cerevisiae and Saccharomyces paradoxus Strains on Ige Production by Mouse Spleen Cells

The genus Saccharomyces includes industrial yeasts that are used for bread and alcoholic beverage production. Saccharomyces strains isolated from natural resources, referred to as “wild” yeasts, are used for making products with strain-specific flavors that are different from those of the “domesticated” industrial yeasts. The physiological effects of wild yeast are poorly understood. In this study, we isolated 2 Saccharomyces cerevisiae strains (S02 − 03) and 5 Saccharomyces paradoxus strains (P01 − 02, S01, S04 − 05) from natural resources in the Kiso area and investigated the effect of these fungal strains on IgE production by mouse spleen cells. Culturing spleen cells with heat-killed yeasts resulted in elevated IFN-γ and IL-12 levels followed by significant reduction in IgE levels. The S03 and P01 strains induced IL-12 p40 and IL-10 expression in RAW264 cells. Thus, wild strains of S. cerevisiae and S. paradoxus regulate macrophage cytokine production to improve the Th1/Th2 immune balance and suppress IgE production.

Tannic acid, a higher galloylated pentagalloylglucose, suppresses antigen-specific IgE production by inhibiting ɛ germline transcription induced by STAT6 activation

Interleukin (IL)-4 is a critical stimulator that induces ɛ germline transcripts (ɛGT) for switch recombination to initiate immunoglobulin (Ig) E and is important in allergic disease pathogenesis. We found pentagalloylglucose (PGG) inhibited IL-4-induced ɛGT expression. PGG exerted its inhibitory function by suppressing IL-4-induced activation of IL-4Rα, JAK3 and STAT6. Furthermore, tannic acid, a higher galloylated PGG, attenuated ovalbumin-induced IgE production in vivo by inhibiting IL-4-induced ɛGT expression and the IL-4 signaling pathway. In conclusion, our results suggest that tannic acid may attenuate allergic diseases by suppressing IgE production by inhibiting IL-4-induced signaling.

Minocycline Blocks Asthma-associated Inflammation in Part by Interfering with the T Cell Receptor-Nuclear Factor κB-GATA-3-IL-4 Axis without a Prominent Effect on Poly(ADP-ribose) Polymerase

Minocycline protects against asthma independently of its antibiotic function and was recently reported as a potent poly(ADP-ribose) polymerase (PARP) inhibitor. In an animal model of asthma, a single administration of minocycline conferred excellent protection against ovalbumin-induced airway eosinophilia, mucus hypersecretion, and Th2 cytokine production (IL-4/IL-5/IL-12(p70)/IL-13/GM-CSF) and a partial protection against airway hyperresponsiveness. These effects correlated with pronounced reduction in lung and sera allergen-specific IgE. A reduction in poly(ADP-ribose) immunoreactivity in the lungs of minocycline-treated/ovalbumin-challenged mice correlated with decreased oxidative DNA damage. The effect of minocycline on PARP may be indirect, as the drug failed to efficiently block direct PARP activation in lungs of N-methyl-N'-nitro-N-nitroso-guanidine-treated mice or H(2)O(2)-treated cells. Minocycline blocked allergen-specific IgE production in B cells potentially by modulating T cell receptor (TCR)-linked IL-4 production at the mRNA level but not through a modulation of the IL-4-JAK-STAT-6 axis, IL-2 production, or NFAT1 activation. Restoration of IL-4, ex vivo, rescued IgE production by minocycline-treated/ovalbumin-stimulated B cells. IL-4 blockade correlated with a preferential inhibition of the NF-κB activation arm of TCR but not GSK3, Src, p38 MAPK, or ERK1/2. Interestingly, the drug promoted a slightly higher Src and ERK1/2 phosphorylation. Inhibition of NF-κB was linked to a complete blockade of TCR-stimulated GATA-3 expression, a pivotal transcription factor for IL-4 expression. Minocycline also reduced TNF-α-mediated NF-κB activation and expression of dependent genes. These results show a potentially broad effect of minocycline but that it may block IgE production in part by modulating TCR function, particularly by inhibiting the signaling pathway, leading to NF-κB activation, GATA-3 expression, and subsequent IL-4 production.

Topical application of Taglisodog-eum inhibits the development of experimental atopic dermatitis

Aim of studyTaglisodog-eum (▪, Tuo Li Xiao Du Yin), a standardized herbal formula, has been widely used to modulate diverse carbuncles in oriental medicine. However, it is still unclear whether Taglisodog-eum (TSE) can exert a beneficial role in dermatological disease. In this study, we examined the effect of topical application of TSE on experimental atopic dermatitis (AD) and elucidated its action mechanism. Materials and methodsTo test the effect of TSE treatment on IgE production in vitro, U266B1 cells and primary CD19+ B cells isolated from AD-induced mice were treated with TSE under LPS/IL-4 stimulation and then IgE level in the culture supernatant was measured by ELISA. To evaluate the effect of TSE treatment on the production of AD related pathogenic cytokines, CD4+ T cells isolated from AD-induced mice were treated with TSE under PMA/ionomycin stimulation, then the level of cytokine expression was analyzed by quantitative RT-PCR and ELISA. The effects of TSE on the NFκB promoter activity in T cells and on the expression level of Aicda (activation-induced cytidine deaminase) in B cells were examined. To further examine the in vivo efficacy of TSE on AD progression, TSE was topically applied to ears of mice with atopic dermatitis induced by painting of DNCB and house dust mite extract. AD Progression was estimated by following criteria: (a) ear thickness, clinical score, (b) serum total IgE and mite specific IgE level by ELISA, (c) histological examination of ear tissue by H&E staining and (d) cytokine profile of total ear cells and draining lymph node CD4+ T cells by quantitative real time PCR and ELISA. ResultsTreatment of TSE to the U266B1 cell line and primary CD19+ B cells isolated from AD-induced mice inhibited IgE production. Treatment of TSE down-regulated the expression of several cytokines (IL-4, IL-10, IL-13, IL-17, TNF-α and IFN-γ) in CD4+ T cells isolated from AD-induced mice. Topical application of TSE on the ears of AD-induced mice decreased the severity and progression of disease by reducing ear thickness, clinical scores including dryness, edema. TSE treatment reduced the infiltration of lymphocytes to the inflamed site analyzed by histological evaluation. TSE treatment also decreased serum IgE level and expression of AD-associated pathogenic cytokines (IL-4, IL-5 and IL-13) in total ear cells and dLN CD4+ T cells by inhibiting the translocation of NFκB into nucleus. ConclusionsOur study indicates that protective effect of Taglisodog-eum (TSE) in experimental atopic dermatitis is mediated by inhibiting IgE production and the levels of Th2 type cytokines, suggesting the beneficial effect of TSE on modulating atopic dermatitis.

6b,11b-Dihydroxy-6b,11b-dihydro-7H-indeno[1,2-b]naphtho[2,1-d]furan-7-one (DHFO), a small molecule targeting NF-κB, demonstrates therapeutic potential in immunopathogenic chronic inflammatory conditions

6b,11b-Dihydroxy-6b,11b-dihydro-7H-indeno[1,2-b]naphtho[2,1-d]furan-7-one (DHFO), an easily synthesisable, orally bioavailable and relatively non-toxic small molecule synthesised in our lab, was previously reported to possess anti-oxidant, 5-lipoxygenase inhibitory, anti-inflammatory and peripheral analgesic activities. The present work deals with exploration of DHFO's efficacy in immunopathogenic chronic inflammatory conditions — arthritis and allergy. In carrageenan-induced inflammatory air pouch, which resembles the arthritic synovium, DHFO effectively reduced inflammatory redness and swelling and neutrophil infiltration. In complete Freund's adjuvant-induced arthritis, DHFO significantly decreased paw oedema and nitrite levels with efficacy comparable to diclofenac. DHFO inhibited neutrophil activation (observed as decreased myeloperoxidase levels), in both the in vivo models of inflammation. Interestingly, DHFO did not ulcerate the gastrointestinal tract, while diclofenac was observed to be extremely ulcerogenic. In antigen-induced active and passive anaphylaxis (allergy) models, DHFO dose-dependently prevented mesenteric mast cell (MC) degranulation with efficacy comparable to ketotifen. DHFO also inhibited compound 48/80 (C48/80)-induced paw oedema and peritoneal MC degranulation. DHFO stabilised p815 murine MCs stimulated by C48/80 and calcium ionophore—A23187, indicating an action downstream of calcium mobilisation. DHFO's anti-allergic mechanism could be two-pronged involving (1) inhibition of IgE production and/or (2) MC stabilisation. DHFO inhibited lipopolysaccharide (LPS)-induced pro-inflammatory mediator release (ROS, NO, IL-6 levels) and COX2 expression in RAW264.7 murine macrophages. Protein expression studies confirmed DHFO's ability to reduce nuclear levels of NF-κB in LPS-stimulated macrophages. Thus, DHFO is a promising non-ulcerogenic synthetic small molecule lead for immunopathogenic chronic inflammatory conditions.

Immunomodulatory effect of water soluble extract separated from mycelium of Phellinus linteus on experimental atopic dermatitis

BackgroundComplementary and alternative medicine (CAM) is becoming a popular treatment for modulating diverse immune disorders. Phellinus linteus (P. linteus) as one of the CAMs has been used to modulate cancers, inflammation and allergic activities. However, little evidence has been shown about its underlying mechanism of action by which it exerts a beneficial role in dermatological disease in vivo. In this study, we examined the immunomodulatory effects of P. linteus on experimental atopic dermatitis (AD) and elucidated its action mechanism.MethodsThe immunomodulatory effect of total extract of P. linteus on IgE production by human myeloma U266B1 cells was measured by ELISA. To further identify the effective components, P. linteus was fractionated into methanol soluble, water soluble and boiling water soluble extracts. Each extract was treated to U266B1 cells and primary B cells to compare their inhibitory effects on IgE secretion. To test the in vivo efficacy, experimental atopic dermatitis (AD) was established by alternative treatment of DNCB and house dust mite extract into BALB/c mice. Water soluble extract of P. linteus (WA) or ceramide as a positive control were topically applied to ears of atopic mouse every day for 2 weeks and progression of the disease was estimated by the following criteria: (a) ear thickness, clinical score, (b) serum total IgE, IgG and mite specific IgE level by ELSIA, (c) histological examination of ear tissue by H&E staining and (d) cytokine profile of total ear cells and CD4+ T cells by real time PCR and ELSIA.ResultsTreatment of total extracts of P. linteus to U266B1 inhibited IgE secretion. Among the diverse extracts of P. linteus, water soluble extract of P. linteus (WA) significantly reduced the IgE production in primary B cells and B cell line U266B1. Moreover, treatment of WA reduced AD symptoms such as ear swelling, erythema, and dryness and decreased recruitment of lymphocyte into the inflamed site. Interestingly WA treatment significantly reduced IgE level without affecting IgG levels and also down-regulated the levels of pathogenic cytokines (IL-4, IL-13, IL-12 and IFN-γ) and chemokines (CCL17 and CCL22) involved in AD development.ConclusionsOur study indicates that protective effect of water soluble extract of P. linteus in atopic dermatitis is mediated by inhibiting IgE production and expression of AD associated pathogenic cytokines as well as chemokines, suggesting the beneficial effect of P. linteus to modulate allergic skin disease.

Treatment of Atopic Dermatitis by Bone Marrow-derived Clonal Mesenchymal Stem Cells (119.3)

Abstract Mesenchymal stem cells (MSCs) can perform immunomodulatory activities, including suppression of T- and B-cell proliferation. MSCs have been primarily studied for their suppression of pro-inflammatory Th1 responses, but their therapeutic effects on atopic dermatitis (AD), which is a Th2-dominant disease, have not been reported. In this study, using an ovalbumin-induced mouse AD model, we investigated whether MSCs reduce the allergic inflammation in AD. We used mouse bone marrow-derived clonal mesenchymal stem cells (mcMSCs) isolated by our newly established isolation protocol termed the subfractionation culturing method (SCM). When injected intravenously into animals between antigen challenges, mcMSCs were found to inhibit the majority of AD-specific pathological changes by decreasing cell infiltration in skin lesions and reducing serum level of IgE. To explore the mechanism of this effect, we investigated the effects of mcMSCs on IgE production in B cells. We found that mcMSCs significantly inhibited IgE production when B cells were stimulated either by lipopolysaccharide (LPS) or anti-CD40 antibodies in the presence of interleukin (IL)-4. In addition, allogeneic mcMSCs can inhibit both isotype class switching and terminal differentiation of B cells probably by inhibiting NF-κB activation. These results suggest that mcMSC-based cell therapy is a promising therapeutic modality for AD treatment.