Jung-Ho Sohn

Acute exposure to silica nanoparticles aggravate airway inflammation: different effects according to surface characteristics.

Silica nanoparticles (SNPs) are widely used in many scientific and industrial fields despite the lack of proper evaluation of their potential toxicity. This study examined the effects of acute exposure to SNPs, either alone or in conjunction with ovalbumin (OVA), by studying the respiratory systems in exposed mouse models. Three types of SNPs were used: spherical SNPs (S-SNPs), mesoporous SNPs (M-SNPs), and PEGylated SNPs (P-SNPs). In the acute SNP exposure model performed, 6-week-old BALB/c female mice were intranasally inoculated with SNPs for 3 consecutive days. In the OVA/SNPs asthma model, the mice were sensitized two times via the peritoneal route with OVA. Additionally, the mice endured OVA with or without SNP challenges intranasally. Acute SNP exposure induced significant airway inflammation and airway hyper-responsiveness, particularly in the S-SNP group. In OVA/SNPs asthma models, OVA with SNP-treated group showed significant airway inflammation, more than those treated with only OVA and without SNPs. In these models, the P-SNP group induced lower levels of inflammation on airways than both the S-SNP or M-SNP groups. Interleukin (IL)-5, IL-13, IL-1β and interferon-γ levels correlated with airway inflammation in the tested models, without statistical significance. In the mouse models studied, increased airway inflammation was associated with acute SNPs exposure, whether exposed solely to SNPs or SNPs in conjunction with OVA. P-SNPs appear to be relatively safer for clinical use than S-SNPs and M-SNPs, as determined by lower observed toxicity and airway system inflammation.

Characterization of the major allergens of Pachycondyla chinensis in ant sting anaphylaxis patients

Background The ant species Pachycondyla chinensis, which has spread from Far Eastern Asia to New Zealand and North America, induces anaphylactic reactions in human with its sting. However, the major allergens of P. chinensis have not yet been characterized.

Characterization of Buckwheat 19-kD Allergen and Its Application for Diagnosing Clinical Reactivity

Background: The 19-kD protein of buckwheat (BW) has been suggested to be a major allergen, but its characteristics and clinical significance are poorly defined. Methods: cDNA of the 19-kD BW allergen was cloned and expressed in Escherichia coli. Allergenicity and cross-allergenicity were confirmed by inhibition immunoblotting or by ELISA inhibition. The recombinant (r19-kD) protein was assessed for clinical utility in the diagnosis of BW reactivity in 18 BW-allergic and 19 BW-asymptomatic sensitized subjects using receiver operating characteristic analysis. Results: The 19-kD BW allergen, which is composed of 135 amino acids, has a weak homology to the vicilin-like allergens of cashew (Ana o 1), English walnut (Jug r 2) and 7 S globulin from Sesamum indicum. The r19-kD protein can inhibit sIgE binding to native 19-kD BW allergen. The maximum percentage inhibition of sIgE binding to crude BW extract was 56%. About 83.3% of the BW allergy patients had sIgE bound to r19-kD protein, compared to only 1 of the 19 BW-asymptomatic sensitized subjects. The areas under the receiver operating characteristic curves for the skin prick tests [0.925 (95% confidence interval: 0.839–1.012), p < 0.001] as well as r19-kD protein sIgE ELISAs [0.860 (95% confidence interval: 0.725–0.995), p <0.001] were higher than that of BW sIgE coated allergen particle test results [0.803 (95% confidence interval: 0.661–0.945), p = 0.002]. Conclusions: The 19-kD BW allergen may be the major allergen from BW. For the diagnosis of clinical reactivity to BW, the r19-kD protein sIgE ELISA test was more discriminative than the coated allergen particle sIgE measurement using whole BW extract.

Optimal conditions for the removal of house dust mite, dog dander, and pollen allergens using mechanical laundry

BACKGROUND Mechanical laundry is an effective tool for the environmental control of allergens, but the optimal conditions for removing allergens are not yet clear. OBJECTIVE To evaluate the optimal conditions of mechanical laundry for the removal of house dust mite (HDM), dog dander, and pollen allergens. METHODS The 4 washing modes of 30 degrees C (86 degrees F), 40 degrees C (104 degrees F), 60 degrees C (140 degrees F), and steam water (SW) with detergent were evaluated. Allergen removal performance was assayed using a 2-site enzyme-linked immunosorbent assay (ELISA) or an ELISA inhibition test. RESULTS Using the 30 degrees C and 40 degrees C washing modes, only 6.5% and 9.6% of Dermatophagoides farinae, respectively, were killed. However, using the 60 degrees C and SW washing modes, all HDMs were killed. The amounts of Der f 1 remaining after the 30 degrees C, 40 degrees C, 60 degrees C, and SW washing modes were 26.8%, 2.4%, 1.3%, and 0.6%, respectively, with unmanipulated contaminated sheets. The effects of rinse on Der f 1 levels after the 30 degrees C washing were greater compared with those after the 40 degrees C, 60 degrees C, and SW modes. The amounts of Can f 1 in the extractions after washing were 0.3% to 1.3% for all modes, and all extracts, even without a rinse, did not inhibit specific IgE binding to dog allergens according to ELISA. The remaining pollen allergen levels after washing were lower in the 60 degrees C and SW modes than in the lower temperature modes. However, the levels did not differ among the various washing modes after rinsing once. CONCLUSION Water temperature and number of rinses are critical factors for the removal of HDM, dog dander, and pollen allergens.

Obesity Increases Airway Hyperresponsiveness via the TNF-α Pathway and Treating Obesity Induces Recovery

Obesity is a known risk factor for allergic asthma. It has been recognized as a key player in the pathogenesis of several inflammatory disorders via activation of macrophages, which is also vital to the development of allergic asthma. We investigated the mechanism of obesity-related asthma and whether treating obesity through exercise or diet ameliorates the severity of asthma in the obesity-related asthma model. We generated diet-induced obesity (DIO) in C57BL/6 mice by high-fat-feeding and ovalbumin-induced asthma (lean-OVA or DIO-OVA). The DIO-OVA mice were then treated with tumor necrosis factor (TNF)-α neutralizing antibody as a TNF-α blockade or a Cl2MDP-containing liposome to induce an alveolar macrophage deficiency. To treat obesity, the DIO-OVA mice were under dietary restrictions or exercised. The pathophysiological and immunological responses were analyzed. Airway hyperresponsiveness (AHR), serum IgE and TNF-α levels in the lung tissue increased in the DIO-OVA mice compared to the lean-OVA mice. Both the TNF-α blockade and depletion of alveolar macrophages in the DIO-OVA mice decreased AHR compared to the DIO-OVA mice. Treating obesity by exercise or through dietary means also reduced pulmonary TNF-α levels and AHR in the DIO-OVA mice. These results suggest that restoring normal body weight is an appropriate strategy for reducing TNF-α levels, and controlling inflammation may help improve asthma severity and control in obesity-related asthma.

Cell permeable NFAT inhibitory peptide Sim-2-VIVIT inhibits T-cell activation and alleviates allergic airway inflammation and hyper-responsiveness

Nuclear factor of activated T cells (NFAT) is an important transcription factor for the production of interleukin (IL)-2 upon T-cell receptor (TcR) signaling. Therefore, inhibition of the NFAT-carcineurin pathway is an important target for inflammatory disease inhibition and graft rejection. A novel cell permeable peptide (CPP), Sim-2, has been identified from a human transcription factor, and Sim-2-CPP conjugated to β-galactosidase or EGFP protein was efficiently delivered into cells in vitro and in vivo. A cell permeable form of the NFAT inhibitory peptide VIVIT (Sim-2-VIVIT) was synthesized and showed inhibitory effects on human CD4 or CD8 T-cell activation through NFAT transcriptional activity suppression and IL-2 inhibition. Intranasal administration of the Sim-2-VIVIT peptide in an ovalbumin (OVA)-induced murine asthma model alleviated peribronchial and perivascular infiltration of inflammatory cells in the lung and caused airway remodeling and airway hyper-responsiveness. These results suggest that cell permeable Sim-2-VIVIT peptide has clinical potential as an immunosuppressive agent for inflammatory diseases.

Application of the 16-kDa buckwheat 2 S storage albumin protein for diagnosis of clinical reactivity

BACKGROUND The 16-kDa protein of buckwheat (BW) has been implicated as a major allergen in BW allergy. OBJECTIVE To characterize the 16-kDa allergen and evaluate its clinical significance as an indicator of BW allergy. METHODS Complementary DNA from the 16-kDa allergen was cloned and expressed in Escherichia coli. Allergenicity was confirmed with IgE immunoblotting or with an enzyme-linked immunosorbent assay. The clinical utility of the recombinant protein (r16 kDa) for diagnosis of BW reactivity was evaluated in 18 BW-allergic and in 20 asymptomatic BW-sensitized subjects. RESULTS The 16-kDa allergen, composed of 127 amino acids, has 50% homology to the reported 8-kDa BW allergen, which belongs to the 2 S storage albumin. The r16-kDa protein can inhibit specific IgE (sIgE) antibody binding to the native BW 16-kDa allergen but minimally inhibited sIgE binding to crude BW extract. Approximately 77.8% of patients with the BW allergy produced sIgE antibodies to the r16-kDa protein, compared with a complete lack of reactivity in the 20 asymptomatic BW-sensitized subjects. The areas of the receiver operating characteristic curves for the skin prick test (mean, 0.93; 95% confidence interval, 0.85 to approximately 1.01; P < .001) and the rl6-kDa enzyme-linked immunosorbent assay (mean, 0.93; 95% confidence interval, 0.84 to approximately 1.01; P < .001) were higher than the area of the BW IgE measurement curve determined by ImmunoCAP (a system for assaying serum IgE) (mean, 0.80; 95% confidence interval, 0.66 to approximately 0.94; P = .002). CONCLUSIONS The 16-kDa allergen belongs to the 2 S storage albumin. Measurement of rl6-kDa sIgE was more discriminating than measurement of ImmunoCAP sIgE in whole BW extracts for the diagnosis of clinical reactivity to BW.

A novel human anti-VCAM-1 monoclonal antibody ameliorates airway inflammation and remodelling

Asthma is a chronic inflammatory disease induced by Type 2 helper T cells and eosinophils. Vascular cell adhesion molecule‐1 (VCAM‐1) has been implicated in recruiting eosinophils and lymphocytes to pathological sites in asthma as a regulatory receptor. Accordingly, monoclonal antibody (mAb) against VCAM‐1 may attenuate allergic inflammation and pathophysiological features of asthma. We attempted to evaluate whether a recently developed human anti‐VCAM‐1 mAb can inhibit the pathophysiological features of asthma in a murine asthma model induced by ovalbumin (OVA). Leucocyte adhesion inhibition assay was performed to evaluate the in vitro blocking activity of human anti‐VCAM‐1 mAb. OVA‐sensitized BALB/c mice were treated with human anti‐VCAM‐1 mAb or isotype control Ab before intranasal OVA challenge. We evaluated airway hyperresponsiveness (AHR) and bronchoalveolar lavage fluid analysis, measured inflammatory cytokines and examined histopathological features. The human anti‐VCAM‐1 mAb bound to human and mouse VCAM‐1 molecules and inhibited adhesion of human leucocytes in vitro. AHR and inflammatory cell counts in bronchoalveolar lavage fluid were reduced in mice treated with human anti‐VCAM‐1 mAb as compared with a control Ab. The levels of interleukin (IL)‐5 and IL‐13, as well as transforming growth factor‐β, in lung tissue were decreased in treated mice. Human anti‐VCAM‐1 mAb reduced goblet cell hyperplasia and peribronchial fibrosis. In vivo VCAM‐1 expression decreased in the treated group. In conclusion, human anti‐VCAM‐1 mAb attenuated allergic inflammation and the pathophysiological features of asthma in OVA‐induced murine asthma model. The results suggested that human anti‐VCAM‐1 mAb could potentially be used as an additional anti‐asthma therapeutic medicine.

Endotoxin Is Not Essential for the Development of Cockroach Induced Allergic Airway Inflammation

Purpose Cockroach (CR) is an important inhalant allergen and can induce allergic asthma. However, the mechanism by which CR induces airway allergic inflammation and the role of endotoxin in CR extract are not clearly understood in regards to the development of airway inflammation. In this study, we evaluated whether endotoxin is essential to the development of CR induced airway allergic inflammation in mice. Materials and Methods Airway allergic inflammation was induced by intranasal administration of either CR extract, CR with additional endotoxin, or endotoxin depleted CR extract, respectively, in BALB/c wild type mice. CR induced inflammation was also evaluated with toll like receptor-4 (TLR-4) mutant (C3H/HeJ) and wild type (C3H/HeN) mice. Results Intranasal administration of CR extracts significantly induced airway hyperresponsiveness (AHR), eosinophilic and neutrophilic airway inflammation, as well as goblet cell hyperplasia in a dose-dependent manner. The addition of endotoxin along with CR allergen attenuated eosinophilic inflammation, interleukin (IL)-13 level, and goblet cell hyperplasia of respiratory epithelium; however, it did not affect the development of AHR. Endotoxin depletion in CR extract did not attenuate eosinophilic inflammation and lymphocytosis in BAL fluid, AHR and IL-13 expression in the lungs compared to CR alone. The attenuation of AHR, eosinophilic inflammation, and goblet cell hyperplasia induced by CR extract alone was not different between TLR-4 mutant and the wild type mice. In addition, heat inactivated CR extract administration induced attenuated AHR and eosinophilic inflammation. Conclusion Endotoxin in CR extracts may not be essential to the development of airway inflammation.