Serum OVA-specific IgE Research Articles

Effect of tangeretin on ovalbumin-provoked allergic respiratory asthma in Swiss albino mice

Purpose: To evaluate the effect of tangeretin on ovalbumin (OVA)-induced allergic respiratory asthma in Swiss albino mice. Methods: Mice were injected intraperitoneally with 50 μg OVA dissolved in water and 1 mg of aluminium hydroxide (alum) on the first day, and on day 12. On days 22, 26 and 30, the mice were exposed to 1 % OVA for 30 min. Mice in the asthma control group received saline via the same route. Tangeretin (50 or 100 mg/kg body weight) was given orally from day 1 to day 30. On the day of sensitization and challenge, tangeretin (50 or 100 mg/kg body weight) was injected 30 min prior to the treatment. Bronchial function, degree of inflammation, broncho-alveolar lavage fluid (BALF), levels of IL-4, IL-5, IL-13 and IFN-γ in BALF, serum total IgE, OVA-specific IgE, IgG1 and IgG2a, as well as histopathology of lungs were determined. Results: Tangeretin administration led to significant reduction in airway sensitivity, and significant decreases in leukocyte, macrophage, and eosinophil counts (p < 0.05). It also brought about significant decreases in the levels of the pro-inflammatory cytokines (IL-4, IL-5 and IL-13) in both BALF and serum (p < 0.05). In addition, tangeretin reversed the OVA-induced histopathological anomalies in the lungs. Conclusion: These results demonstrate that tangeritin exerts protective effects against OVA-induced allergic respiratory asthma in Swiss albino mice, and that the drug can potentially be developed for the management of allergic asthma. Keywords: Allergic asthma, Inflammation, Cytokines, Ovalbumin, Tangeretin

The optimal age for epicutaneous sensitization following tape-stripping in BALB/c mice

BackgroundDirect contact of food proteins with eczematous lesions is thought to be the main cause of epicutaneous sensitization. To further investigate the development and pathogenesis of food allergy in vivo, a good mouse model of epicutaneous sensitization is needed. However, a fundamental problem in that regard is that the optimal age for epicutaneous sensitization of mice is unknown. In this study, we attempted to elucidate that optimal age. MethodsDorsal skin of wild-type BALB/c female mice (1, 3, 8 and 24 weeks old) was shaved, depilated and tape-stripped. A Finn chamber containing a 20-μl-aliquot of 20-mg/ml (OVA) was applied to the tape-stripped skin on 3 consecutive days/week, for 3 weeks. The body temperature was measured after intraperitoneal OVA challenge. Serum OVA-specific IgE titers and OVA-induced cytokine production by spleen cells were measured by ELISA. Dendritic cells (DCs) that migrated to the draining lymph nodes were quantified by FITC-labeled OVA and flow cytometry. The mRNA expression levels in the dorsal skin were measured by qPCR. ResultsA significant age-dependent body temperature decline was observed after OVA challenge. The serum OVA-specific IgE titer, OVA-induced cytokine production (i.e., IL-4, IL-5 and IL-13) by spleen cells, and number of FITC-OVA-engulfing DCs increased with age. In addition, mRNA for IL-33, but not TSLP or IL-25, was significantly induced in the skin by tape-stripping and increased with age. ConclusionsTwenty-four-week-old mice showed the greatest DC migration, Th2 polarization, IgE production and body temperature decline. Skin-derived IL-33 is likely to play key roles in those changes.

Fine particulate air pollution epigenetically affects TET1 expression in dendritic cells and exacerbates the inflammation of Allergic Rhinitis

Prevalence of Allergic Rhinitis(AR) is increasing year by year. It has been proved that environmental factors and epigenetic regulation are involved in the differentiation and development of immunocytes in AR. We aim to clarify whether fine particulate air pollution (for example, PM2.5) affects ten-eleven-translocation 1 (TET1) 5-mC dioxygenase enzymes in the differentiation of dentritic cells(DC) in AR. Thirty-four HDM-sensitized patients with AR and twenty healthy controls were enrolled to compare the expression of TET1 in the peripheral blood mononuclear cells(PBMCs) and DC using quantitative reverse transcription polymerase chain reaction(qRT-PCR), Western blot and flow cytometry. In vitro, monocyte-derived DCs(moDCs) exposed to PM2.5 were detected by flow cytometry to evaluate the expression costimulatory molecules. Further to co-culture DC with CD4+ T cell and evaluate the number and immunosuppressive function of activated regulatory T cell(aTreg, CD4+CD25+CD45RAlowFoxp3high) by CFSE. In vivo, BALB/c mice were OVA-sensitized and challenged with or without PM2.5 exposed. HE and PAS staining in lung tissue, cytokines in bronchoalveolar lavage fluid, serum OVA-specific IgG1 and IgE were observed. TET1 expression in lung was evaluated by qRT-PCR and flow cytometry. TET1 expression decreases in PBMC of AR patients, especially in DC. MoDCs exposed to PM2.5 highly express CD86 and downregulate the number and immunosuppressive function of aTreg, increasing Th2 polarization and decreasing TET1 expression. Exacerbating effect of PM2.5 in OVA-sensitized and challenged mice decreases the expression of TET1. Exposing to fine particulate air pollution epigenetically downregulates TET1 expression in dendritic cells and exacerbates the inflammation of Allergic Rhinitis.

The crystal structure of titanium dioxide nanoparticles influences immune activity in vitro and in vivo

BackgroundThe use of engineered nanoparticles (NP) is widespread and still increasing. There is a great need to assess their safety. Newly engineered NP enter the market in a large variety; therefore safety evaluation should preferably be in a high-throughput fashion. In vitro screening is suitable for this purpose. TiO2NP exist in a large variety (crystal structure, coating and size), but information on their relative toxicities is scarce. TiO2NP may be inhaled by workers in e.g. paint production and application. In mice, inhalation of TiO2NP increases allergic reactions. Dendritic cells (DC) form an important part of the lung immune system, and are essential in adjuvant activity. The present study aimed to establish the effect of a variety of TiO2NP on DC maturation in vitro. Two NP of different crystal structure but similar in size, uncoated and from the same supplier, were evaluated for their adjuvant activity in vivo.MethodsImmature DC were differentiated in vitro from human peripheral blood monocytes. Exposure effects of a series of fourteen TiO2NP on cell viability, CD83 and CD86 expression, and IL-12p40 and TNF-α production were measured. BALB/c mice were intranasally sensitized with ovalbumin (OVA) alone, OVA plus anatase TiO2NP, OVA plus rutile TiO2NP, and OVA plus Carbon Black (CB; positive control). The mice were intranasally challenged with OVA. OVA-specific IgE and IgG1 in serum, cellular inflammation in bronchoalveolar lavage fluid (BALF) and IL-4 and IL-5 production in draining bronchial lymph nodes were evaluated.ResultsAll NP dispersions contained NP aggregates. The anatase NP and anatase/rutile mixture NP induced a higher CD83 and CD86 expression and a higher IL-12p40 production in vitro than the rutile NP (including coated rutile NP and a rutile NP of a 10-fold larger primary diameter). OVA-specific serum IgE and IgG1 were increased by anatase NP, rutile NP, and CB, in the order rutile<anatase<CB. The three particles similarly increased IL-4 and IL-5 production by bronchial LN cells and eosinophils and lymphocytes in the BALF. Neutrophils were induced by rutile NP and CB but not by anatase NP.ConclusionsOur data show that measuring CD83 and CD86 expression and IL-12p40 and TNF-α production in DC in vitro may provide an efficient way to screen NP for potential adjuvant activity; future studies should establish whether this also holds for other NP. Based on antigen-specific IgE and IgG1, anatase NP have higher adjuvant activity than rutile NP, confirming our in vitro data. Other parameters of the allergic response showed a similar response for the two NP crystal structures. From the viewpoint of safe(r) by design products, rutile NP may be preferred over anatase NP, especially when inhalation exposure can be expected during production or application of the product.

Maternal Separation as Early-Life Stress Causes Enhanced Allergic Airway Responses by Inhibiting Respiratory Tolerance in Mice.

Epidemiologic studies indicate that exposure to psychosocial stress in early childhood is a risk factor of adult-onset asthma, but the mechanisms of this relationship are poorly understood. Therefore, we examined whether early-life stress increases susceptibility to adult-onset asthma by inhibiting the development of respiratory tolerance. Neonatal BALB/c female mice were aerosolized with ovalbumin (OVA) to induce immune tolerance prior to immune sensitization with an intraperitoneal injection of OVA and the adjuvant aluminum hydroxide. Maternal separation (MS) was applied as an early-life stressor during the induction phase of immune tolerance. The mice were challenged with OVA aerosol in adulthood, and allergic airway responses were evaluated, including airway hyper-responsiveness to inhaled methacholine, inflammatory cell infiltration, bronchoalveolar lavage fluid levels of interleukin (IL)-4, IL-5, and IL-13, and serum OVA-specific IgE. We then evaluated the effects of MS on the development of regulatory T (Treg) cells in bronchial lymph nodes (BLN) and on splenocyte proliferation and cytokine expression. In mice that underwent MS and OVA tolerization, the allergic airway responses and OVA-induced proliferation and IL-4 expression of splenocytes were significantly enhanced. Furthermore, exposure to MS was associated with a lower number of Treg cells in the BLN. These findings suggest that exposure to early-life stress prevents the acquisition of respiratory tolerance to inhaled antigen due to insufficient Treg cell development, resulting in Th2-biased sensitization and asthma onset. We provide the evidence for inhibitory effects of early-life stress on immune tolerance. The present findings may help to clarify the pathogenesis of adult-onset asthma.

고려엉겅퀴 에탄올 추출물의 대식세포에서의 항염효능과 정상 및 ovalbumin 감작 Balb/c에서 혈청 면역글로불린에 대한 효능

This study evaluated the anti-inflammatory effects of Cirsium setidens ethanol extract on raw 264.7 cells and effect on serum immunoglobulins in normal and ovalbumin-sensitized Balb/c mouse. In animal experiments, the Balb/c mice were randomly divided into four to five groups, which may or may not be ovalbumin-sensitized. Mice were administered AIN-93G basal diet (for control groups, NC or SC) or AIN-93G diet mixed with 2.5%(G-2.5), 5%(G-5), and 10%(G-10) Cirsium setidens extracts (treated groups) for 7 weeks. The serum IgE and IgG1 levels and the degree of oxidative DNA damage were evaluated. The results showed that the NO release from raw cell was not significantly changed by the Cirsium setidens ethanol extract, whereas the levels of TNF-α and IL-1β were significantly decreased from 500 and 250 μg/mL, respectively. In non-sensitized animal experiments, the serum IgE concentration of the experimental groups was dose-dependently reduced compared with the control group (P0.05). The tail length and moment resulting from DNA fragmentation of blood white cells decreased significantly in G-5 and G-10 groups. In the OVA-sensitized animal experiment, the serum OVA-specific IgE levels were significantly decreased in groups fed with Cirsium setidens extract. The tail DNA and tail moment in all the experimental groups decreased compared with the SC group. Cirsium setidens is a vegetable with antioxidant, anti-inflammatory and immunomodulating properties, which warrant additional research.

Intranasal immunization with phosphorylcholine suppresses allergic rhinitis in mice.

Intranasal immunization with phosphorylcholine (PC) is known to reduce immunoglobulin (Ig)E production. However, its effects on the occurrence of allergic rhinitis (AR) are unknown. This study was performed to evaluate the effects of PC-keyhole limpet hemocyanin (PC-KLH) and to examine the effects on the occurrence of AR in a murine model of AR. In vivo study using an animal model. Forty-five female BALB/c mice were divided into three groups; those pretreated with intranasal administration of PC-KLH followed by intraperitoneal sensitization and nasal challenge with ovalbumin (OVA) (group A), those untreated with PC-KLH followed by sensitization and nasal challenge with OVA (group B), and those untreated with PC-KLH or OVA as controls (group C). Nasal symptoms, allergic inflammation in the nasal mucosa, OVA specific IgE production, and cytokine profile were compared among those three groups. Dendritic cells (DCs) were isolated from splenic cells and PC-KLH-stimulated interleukin (IL)-12p40 production was measured. The mice pretreated with PC-KLH showed lower allergic nasal symptoms and inflammation compared to untreated mice. The levels of total IgE and OVA-specific IgE in serum, and IL-4 production by nasal and splenic CD4+ T cells were significantly reduced by PC-KLH pretreatment. Furthermore, IL-12p40 production by DCs was induced by PC-KLH in a dose-dependent manner. Intranasal administration of PC-KLH suppressed allergic inflammation in nasal mucosa and antigen-specific IgE production by downregulating Th2-type immune response. Intranasal immunization with PC might be useful to prevent AR and upper airway bacterial infection. NA. Laryngoscope, 128:E234-E240, 2018.

Effects of ovalbumin exposure during pregnancy of mice on the ovalbumin re-exposure in adult progeny

Objective: To observe the immunoreaction of offspring mice by ovalbumin (OVA) re-exposure after their mothers exposed to OVA during pregnancy. Method: A prospective controlled study was conducted to observe mice after repeated OVA exposures at 6-8 weeks.Their mothers were exposed to OVA during different stages of pregnancy.The symptoms were recorded and scored.The levels of OVA-specific IgE in serum, interferon-γ(IFN-γ) and interleukin-4(IL-4) in supernatant of spleen primary lymphocytes in vitro were measured by ELISA.The results were analyzed by single factor analysis of variance or rank sum test. Result: All the mice in each group had acute diarrhea.The diarrhea happened earliest (2 days) and most severe in the late pregnancy group (early pregnancy group 7.0±1.0; middle pregnancy group: 7.1±1.1; late pregnancy group: 9.9±2.2, P<0.01). The levels of absorbance of OVA-specific IgE in the pregnancy groups were higher than those of the control group.The absorbance of OVA-specific IgE in late pregnancy group was the highest (control: 0.27±0.06; early pregnancy group: 0.51±0.13; middle pregnancy group: 0.50±0.09; late pregnancy group: 0.63±0.13, P<0.01). There was no significant change in IFN-γ expression in cultured supernatant of spleen lymphocytes in each group (control: (133±7) pg/ml; early pregnancy group: (133±4) pg/ml; middle pregnancy group: (134±6) pg/ml; late pregnancy group: (132±4) pg/ml, all P value >0.05). The expression of IL-4 in the experimental groups was higher than that in the control group, especially in late pregnancy group(control: (25.3±2.4) pg/ml; early pregnancy group: (32.4±4.4) pg/ml; middle pregnancy group: (35.0±5.4) pg/ml; late pregnancy group: (47.1±5.8) pg/ml; P value all<0.01). Conclusion: The allergic reaction of the OVA re-exposure progeny whose mothers were exposed to OVA in the late pregnancy period was most severe, suggesting that late pregnancy period might be the high risk stage of intrauterine sensitization, or"window period".

Ouabain attenuates ovalbumin-induced airway inflammation.

Ouabain, an Na+/K+-ATPase inhibitor hormone, presents immunomodulatory actions, including anti-inflammatory effect on acute inflammation models. In the present study, the effect of ouabain in a model of allergic airway inflammation induced by ovalbumin (OVA) was assessed. Initially, it was observed that ouabain treatment inhibited cellular migration induced by OVA on bronchoalveolar lavage fluid (BALF), mostly granulocytes, without modulating macrophage migration. In addition, it was observed, by flow cytometry, that ouabain reduces CD3high lymphocytes cells on BALF. Furthermore, treatment with ouabain decreased IL-4 and IL-13 levels on BALF. Ouabain also promoted pulmonary histological alterations, including decreased cell migration into peribronchiolar and perivascular areas, and reduced mucus production in bronchioles regions observed through hematoxylin-eosin (HE) and by periodic acid-Schiff stain, respectively. Allergic airway inflammation is characterized by high OVA-specific IgE serum titer. This parameter was also reduced by the treatment with ouabain. Therefore, our data demonstrate that ouabain negatively modulates allergic airway inflammation induced by OVA.

Rutin has anti-asthmatic effects in an ovalbumin-induced asthmatic mouse model

Purpose: To investigate the anti-asthmatic effects of the flavonoid rutin in an ovalbumin (OVA)-induced asthmatic mouse model. Methods: Asthma was induced by OVA induction. Effects of rutin (25–100 mg/kg BW) on interleukins (IL)-4, IL-5, and IL-13 and interferon (IFN)-γ in bronchoalveolar lavage fluid (BALF) and serum OVAspecific IgE levels were assessed by enzyme linked immunosorbent assay (ELISA). Airway hyperresponsiveness (AHR) was measured by determining lung compliance and airway resistance. Expression levels of p-I-κBα, p-NF-κB p65, TNF-α, and Foxp3 were also assessed. The CD4 + CD25 + Foxp3 + T regulatory cell population was determined by flow cytometry. Results: Rutin significantly ( p < 0.05) decreased Th2 cytokines (IL-4, IL-5, IL-13), whereas it enhanced IFN-γ levels in BALF. Decreased OVA-specific serum IgE was also observed with improved lung histology. Rutin was effective in reducing AHR and regulating the expression levels of p-I-κBα, p-NF-κB p65, and TNF α, and also increased the CD4 + CD25 + Foxp3 + Treg cell population and Foxp3 mRNA and protein levels. Conclusion: Rutin effectively suppresses OVA-induced asthma and improves airway function by suppressing inflammatory mediators and inflammatory cell infiltration. Keywords: Asthma, CD4 + CD25 + , Cytokines, Inflammation, Ovalbumin, Rutin, Protein levels

Immunologic regulatory effects of human umbilical cord blood-derived mesenchymal stem cells in a murine ovalbumin asthma model.

Mesenchymal stem cells (MSCs) have multiple immunomodulatory properties and hold therapeutic potential for inflammatory diseases. However, the therapeutic and immunologic effects of human umbilical cord blood-derived MSCs (huMSCs) remain largely unexamined for asthma. This study was to investigate the immunomodulatory properties of huMSCs in an ovalbumin (OVA)-induced murine asthma model. Mice were injected intraperitoneally with OVA and an aluminium hydroxide adjuvant. huMSCs were administered via the tail vein (5×105 cells/100uL) to female BALB/c mice prior to the initial OVA challenge. The effects of huMSCs were assessed by investigating airway hyperresponsiveness, histological changes, inflammatory cell numbers, serum allergen-specific antibodies, cytokine production in spleen, lung tissue, and bronchoalveolar lavage (BAL) fluid as well as expansion of regulatory T cells. Administration of huMSCs significantly reduced methacholine bronchial hyperresponsiveness and eosinophil counts in BAL cells. Similarly, there was a significant decrease in serum OVA-specific IgE and IgG1 levels along with Th2 cytokine production (IL-4, IL-5, and IL-13) in the lung and spleen tissues, whereas increased percentage of regulatory T cells was observed after treatment with huMSCs. Our results suggest that huMSC treatment reduces OVA-induced allergic inflammation, which could be mediated by regulatory T cells.

Vitamin D supplementation modulates Th2 immune response by inducing T regulatory cells in allergic conjunctivitis

Abstract Allergic conjunctivitis (AC) is one of the most common diseases encountered in eye clinics. Recently, Vitamin D deficiency has been shown to be associated with allergic AC. This present study was designed to evaluate the efficacy of vitamin D to suppress ovalbumin (OVA)-sensitized AC in a murine model. BALB/c mice were sensitized with OVA and aluminum hydroxide. Two weeks later, mice were challenged by OVA eyedrops for 12 days with intraperitoneal injection of vehicle or 1,25(OH)2D3. We evaluated clinical signs, the infiltration of inflammatory cells, regulatory T cells (Treg), serum IgE, and Th2 cytokines through flow cytometry and ELISA. In addition, to evaluate inhibitory function of Treg cells, anti–CD25 blocking antibody or isotype control antibody was injected intravenously during topical OVA challenge. AC development and conjunctival infiltration of eosinophils and mast cells were significantly impaired with 1,25(OH)2D3 treatment. In addition, 1,25(OH)2D3 suppressed production of OVA-specific IgE in serum and Th2 cytokines in vitro T cell assays, such as IL-4 and IL-13, compared to vehicle group. Interestingly, 1,25(OH)2D3 led to increase Treg cells population in draining LNs and suppressive levels of clinical signs and inflammatory cells infiltration into conjunctivae were reversed by depleting Treg cells. Our results suggest that vitamin D supplement alleviate allergic conjunctivitis, indicated by suppressing Th2 response in draining LNs and inflammatory cells infiltration into conjunctiva through increasing population of Treg cells in draining LNs. Therefore our results demonstrated the therapeutic potential of vitamin D for allergic diseases including asthma or atopic dermatitis.

Experimental food-induced anaphylaxis is driven by C5aR1 actvivation on mast cells

Abstract Food-induced anaphylaxis is a serious allergic reaction mainly caused by antigen cross-linking of IgE-loaded Fce-receptors on mast cells (MCs). Such MC activation leads to the release of pro-inflammatory mediators which results in the manifestation of the disease. Here, we demonstrate that the complement fragment C5a plays an important and until now not recognized role in the development and severity of food allergy. We subjected C5aR1−/− and wild-type (wt) Balb/c mice to oral ovalbumin (OVA)-induced anaphylaxis. We observed that 78% of wt but only 9% of C5aR1−/− mice suffered from diarrhea and hypothermia after the 7th oral OVA challenge. Importantly, peritoneal MCs strongly expressed C5aR1 after the 7th oral OVA treatment. OVA-specific serum IgE and MCPT-1 levels were significantly decreased in C5aR1-deficient as compared with wt mice. To directly examine the impact of the C5a/C5aR1 axis on MC function, we generated bone marrow-derived MCs (BMMCs) and tested their activation and degranulation potency in response to FceR1 cross-linking. Degranulation of C5aR1−/− BMMCs was reduced by 50% and IL-6 production was decreased by 35% as compared with wt cells. Also, FceR1 cross-linking markedly upregulated C5aR1 MC expression. Further, in contrast to wt mice, C5aR1−/− mice were largely protected from histamine-induced temperature drop in vivo. In line with this finding, C5aR targeting protected human endothelial cells from histamine-induced barrier dysfunction and disruption of tight junction proteins in vitro. Our findings identify a critical role of the C5a/C5aR1 axis in IgE-mediated oral antigen-induced anaphylaxis. C5aR1 may serve as a novel therapeutic target to suppress the inflammatory response in food-induced anaphylaxis.

Effect of Proparacaine in a Mouse Model of Allergic Rhinitis

Objectives Lidocaine, a local anaesthetic is a treatment option in uncontrolled asthma due to its immunomodulatory effects. In the present study, proparacaine (PPC), a derivative of lidocaine was examined for its therapeutic application in a mouse model of allergic rhinitis.Methods The mice were grouped into 4 groups: control group, allergic rhinitis (AR) group, ciclesonide (CIC) group, and PPC group. Nasal symptom scores, eosinophil counts, goblet cell counts, and mast cells counts in the nasal mucosa were measured. Serum ovalbumin (OVA)-specific immunoglobulin (Ig) E, OVA-specific IgG1, OVA-specific IgG2a, interleukin (IL)-4, IL-5, and cortisol levels were measured.Results Intranasal administration of PPC significantly decreased nasal symptoms, number of eosinophils, goblet cells, and mast cells in the lamina propria of the nasal mucosa. Serum OVA-specific IgE, OVA-specific IgG1, OVA-specific IgG2a was significantly higher in the AR compared with the control group. Serum level of IL-4 was significantly lower in the CIC group and PPC group in comparison with AR group. Serum IL-5 showed no significant difference among all groups. No significant difference in serum cortisol levels was observed among the 4 groups.Conclusion PPC appears to have a therapeutic potential in treatment of allergic rhinitis in a mouse model by reducing eosinophil, goblet cell, and mast cell infiltration in the nasal mucosa.

Assessing the effect of 5-hydroxymethylfurfural on selected components of immune responses in mice immunised with ovalbumin.

5-Hydroxymethylfurfural (5-HMF) is one of the most important products of the Maillard reaction. In recent years, many profitable biological effects of this compound have been demonstrated. This study sought to elucidate the anti-allergic effect of 5-HMF by investigating some selected components of the immune response in BALB/c mice immunised with ovalbumin (OVA). Immunised animals had an increased level of serum total and OVA-specific antibodies when compared to the control (P < 0.01).We found that the OVA-induced increase in serum IgE and OVA-specific IgE were significantly suppressed in the groups treated with 5-HMF (P < 0.05). Moreover, interleukin-4 (IL-4) and interferon gamma (IFN-γ) were significantly reduced in a dose-independent manner when compared to the sensitised group (P < 0.05). 5-HMF inhibited the up-regulation of total and OVA-specific IgE through the suppression of the Th2-type immune response in immunised BALB/c mice. 5-HMF could therefore be a novel therapeutic approach for the prevention of IgE-mediated allergic diseases. © 2017 Society of Chemical Industry.

Bufalin Inhibits the Inflammatory Effects in Asthmatic Mice through the Suppression of Nuclear Factor-Kappa B Activity

Asthma is an inflammatory airway disease characterized by increased infiltration of inflammatory cells into the airways and poor respiratory function. Bufalin is one of the biological ingredients obtained from Chansu. Bufalin was found to possess various pharmacological properties including anti-inflammatory activities. However, the effect of bufalin treatment on asthma has not yet been reported. Therefore, this study aimed to investigate the inhibitory effect of bufalin on asthmatic response in a murine model. A mouse asthma model was developed by ovalbumin (OVA) sensitization and challenge in the BALB/c mice. OVA-specific serum IgE and the levels of interleukin (IL)-4, IL-5, and IL-13 in bronchoalveolar lavage fluid (BALF) were determined by an enzyme-linked immunosorbent assay. Recruitment of inflammatory cells into BALF or lung tissues, and goblet cell hyperplasia were evaluated by histological staining. The expression levels of inhibitory subunit of nuclear factor-kappa B (NF-κB) alpha (IκBα) and phosphorylated p65 protein were measured by Western blot analyses. The results demonstrated that bufalin (5 and 10 mg/kg) markedly attenuated hyperresponsiveness, and strongly suppressed the OVA-induced increases of total inflammatory cells including macrophages, eosinophils, lymphocytes, and neutrophils in BALF. The levels of IL-4, IL-5, and IL-13 in BALF and OVA-specific IgE in serum were significantly reduced by bufalin. Histological staining of lung tissues showed that bufalin reduced inflammatory cell infiltration and goblet cell hyperplasia. The results of Western blotting indicated that bufalin suppressed the IκBα degradation from NF-κB, and reduced the level of phosphorylated p65 protein in the lung tissues. These data suggest that bufalin can exert its anti-inflammatory effects possibly through the inhibition of the NF-κB activity.

Protective effect of Bifidobacterium infantis CGMCC313-2 on ovalbumin-induced airway asthma and β-lactoglobulin-induced intestinal food allergy mouse models.

AIMTo determine whether oral administration of Bifidobacterium infantis CGMCC313-2 (B. infantis CGMCC313-2) inhibits allergen-induced airway inflammation and food allergies in a mouse model.METHODSOvalbumin (OVA)-induced allergic asthma and β-lactoglobulin-induced food allergy mouse models were used in this study. Following oral administration of B. infantis CGMCC313-2 during or after allergen sensitization, histopathologic changes in the lung and intestine were evaluated by hematoxylin and eosin (HE) staining. In the allergic asthma mouse model, we evaluated the proportion of lung-infiltrating inflammatory cells. OVA-specific IgE and IgG1 levels in serum and cytokine levels in bronchoalveolar lavage fluid (BALF) were also assessed. In the food allergy mouse model, the levels of total IgE and cytokines in serum were measured.RESULTSOral administration of B. infantis CGMCC313-2 during or after allergen sensitization suppressed allergic inflammation in lung and intestinal tissues, while the proportion of infiltrating inflammatory cells was significantly decreased in the BALF of allergic asthma mice. Moreover, B. infantis CGMCC313-2 decreased the serum levels of total IgE in food allergy mice, and reductions in IgE and IgG1 were also observed in OVA-induced allergic asthma mice. The expression of interleukin-4 (IL-4) and IL-13 in both serum and BALF was suppressed following the administration of B. infantis CGMCC313-2, while an effect on serum IL-10 levels was not observed.CONCLUSIONB. infantis CGMCC313-2 inhibits the secretion of allergen-induced IgE, IL-4 and IL-13, and attenuates allergic inflammation.

Diospyros kaki calyx inhibits immediate-type hypersensitivity via the reduction of mast cell activation

Context: Diospyros kaki L. (Ebenaceae) fruit is widely distributed in Asia and is known to exert anti-inflammatory and antithrombotic effects. Objective: We evaluated the inhibitory effect of aqueous extract of D. kaki calyx (AEDKC) on mast cell-mediated immediate-type hypersensitivity and underlying mechanism of action. Materials and methods: For in vivo, ovalbumin (OVA)-induced active systemic anaphylaxis (ASA) and immunoglobulin (Ig) E-mediated passive cutaneous anaphylaxis (PCA) models were used. In the ASA, AEDKC (1–100 mg/kg) was orally administered 3 times during 14 days. In the PCA, AEDKC was orally treated 1 h before the antigen challenge. The control drug dexamethasone was used to compare the effectiveness of AEDKC. For in vitro, IgE-stimulated RBL-2H3 cells and primary cultured peritoneal mast cells were used to determine the role of AEDKC (0.01–1 mg/mL). Results: Oral administration of AEDKC dose dependently suppressed rectal temperature decrease and increases in serum histamine, total IgE, OVA-specific IgE, and interleukin (IL)-4 in the ASA. In the PCA, AEDKC reduced Evans blue pigmentation. Compared to dexamethasone (10 mg/kg), AEDKC (100 mg/kg) showed similar inhibitory effects in vivo. AEDKC concentration dependently suppressed the release of histamine and β-hexosaminidase through the reduction of intracellular calcium in mast cells. In addition, AEDKC decreased the expression and secretion of tumour necrosis factor-α and IL-4 by the reduction of nuclear factor-κB. The inhibitory potential of AEDKC (1 mg/mL) was similar with dexamethasone (10 μM) in vitro. Conclusions: We suggest that AEDKC may be a potential candidate for the treatment of mast cell-mediated allergic diseases.

Intradermal Delivery of Antigens Enhances Specific IgG and Diminishes IgE Production: Potential Use for Vaccination and Allergy Immunotherapy.

Skin is protected by a tough but flexible multilayered barrier and is a front line for immune responses against invading particles. For many years now, skin has been a tissue where certain vaccines are injected for the prevention of infectious disease, however, the detailed mechanisms of the skin immune response are not yet well understood. Using thin and small injection needles, we carefully injected OVA into a restricted region of mouse skin, i.e., intradermal (ID), and examined the antibody response in comparison with subcutaneous (SC) injection or epicutaneous patch administration of OVA. Epicutaneous patches induced a high IgE response against OVA, but IgG production was low. High IgG production was induced by both ID and SC injection, moreover, ID injection induced higher IgG production without any adjutants. Furthermore, OVA-specific IgE production was diminished by ID injection. We found that ID injection could efficiently stimulate skin resident DCs, drive Th1-biased conditions and diminish IgE production. The ID injection response was regulated by Langerin+ dermal DCs, because OVA was taken up mainly by these cells and, after transiently deleting them, the IgE response was no longer diminished and IgG1 production was enhanced. We also tested whether ID injection might be an effective allergy treatment by attempting to inhibit ongoing IgE production in mice with experimentally induced high serum IgE levels. Multiple ID injections of OVA were shown to prevent elevation of serum OVA-specific IgE after repeated allergen challenge. In contrast, SC OVA injection could only transiently inhibit the OVA-specific IgE production. These findings indicated that ID injection results in higher induction of antigen-specific IgG, and thus may be useful for vaccine delivery with little or no adjuvant components. Moreover, the observed diminishment of IgE and induction of Th1-biased immune responses suggest that ID may be a useful injection route for allergy immunotherapy.

Differences in allergic inflammatory responses in murine lungs: comparison of PM2.5 and coarse PM collected during the hazy events in a Chinese city

Urban particulate matter (PM) is associated with an increase in asthma. PM2.5 (<PM2.5 μm) and coarse PM (CPM: PM2.5–PM10 μm) were collected from the air in a Chinese city during haze events. The amounts of polycyclic aromatic hydrocarbons (PAHs) were higher in PM2.5 than in CPM. Conversely, microbial elements LPS and β-glucan were much higher in CPM than in PM2.5. Concentrations of Si, Al, Fe, and Ti in CPM were greater than in PM2.5, while Pb, Cu and As concentrations were lower than in PM2.5. When RAW264.7 cells were treated with PM2.5 and CPM, the pro-inflammatory response in the cells was associated with the microbial element levels and attenuated partly by both polymyxin B (PMB) and N-acetylcystein (NAC). The expression of the oxidative stress response gene heme oxygenase1 was associated with PAHs levels. The exacerbating effects of the two-types of PM on murine lung eosinophilia were compared to clarify the role of toxic materials. When BALB/c mice were intratracheally instilled with PM2.5 or CPM (total 0.4 mg) + ovalbumin (OVA), both exacerbated lung eosinophilia along with allergy-relevant biological indicators, such as OVA-specific IgE in serum; enhancement of lung pathology when compared with counterpart samples without OVA. The exacerbating effects were greater in microbial element-rich CPM than in organic chemical-rich PM2.5. These results indicate that microbial elements have more potently exacerbating effects on the development of lung eosinophilia than do organic chemicals. In addition, oxidative stress and transition metals might be associated with the exacerbation of this negative effect.