Antibodies are widely used in medicinal and scientific research due to their ability to bind to a specific antigen. Most often, antibodies are composed of heavy and light chain domains. Under physiological conditions, light chains are produced in excess, as compared to the heavy chain. It is now known that light chains are not silent partners of the heavy chain and can modulate the immune response independently. In this work, the first crystal structure of a light chain dimer originating from mice is described. It represents the light chain dimer of 6A8, a monoclonal antibody specific to the allergen Der f 1. Building on the unexpected occurrence of this kind of dimer, we have demonstrated that this light chain is stable in solution alone. Moreover, enzyme-linked immunosorbent assays (ELISA) have revealed that, when the light chain is not partnered to its corresponding heavy chain, it interacts non-specifically with a wide range of proteins. Computational studies were used to provide insight on the role of the 6A8 heavy chain domain in the specific binding to Der f 1. Overall, this work demonstrates and supports the ongoing notion that light chains can function by themselves and are not silent partners of heavy chains.

This study was carried out to detect house dust mites in houses and to investigate group 1 antigens of Dermatophagoid species in Ordu, Giresun, Trabzon and Rize provinces of the Central and Eastern Black Sea Region.
Dust samples obtained from the beds were subjected to both microscopic and antigenic examination. Samples prepared by the lactic acid method for microscopic examination were evaluated under a light microscope. Antigenic analysis was performed by investigating Der p 1 and Der f 1 belonging to D. pteronyssinus and D. farinae by ELISA test.
90.3% of the dust samples were evaluated positive by microscopic examination (10x, 40x) and 149 mites were detected. D. pteronyssinus 74%, D. farinae 13%, Dermatophagoides spp. growth forms 5%, Cheyletus spp. 1%, E. maynei 1%, C. arcuatus 1%, T. putrescentiae 1%, L. destructor 1% and unidentified mites were detected at the rate of 3% respectively. Der p 1 antigen was detected in 93% and Der f 1 antigen in 84.7%. The highest amount of antigen detected in one gram of powder was 1,272 μg for Der p 1 and 0,482 μg for Der f 1.
No difference was observed between mite species and distribution in the provinces where the study was conducted (p<0.05). Dermatophagoides were found in 93% of the population. The low (4%) rate of storage/food mites is related to the fact that samples were not taken from the floors. Antigen accumulation may be important in the beds since the activity of the mites is observed throughout the year in temperate and humid regions. It is thought that this diagnosis method can be used and can be taken into account in terms of the environments in which sensitive people live.
Orta ve Doğu Karadeniz Bölgesi illerinden Ordu, Giresun, Trabzon ve Rize’de evlerde ev tozu akarlarının tespit edilmesi ve Dermatophagoid türlerden D. pteronyssinus ve D. farinae’ye ait antijenlerinin araştırılması amacıyla yürütülmüştür.
Yataklardan temin edilen toz örnekleri hem mikroskobik hem de antijenik incelemeye tabi tutulmuştur. Mikroskobik inceleme için laktik asit yöntemiyle hazırlanmış örnekler ışık mikroskobunda (10x, 40x) değerlendirilmiştir. Antijenik inceleme D. pteronyssinus ve D. farinae’ye ait Der p 1 ve Der f 1’in ELISA testi ile araştırılmasıyla yapılmıştır.
Mikroskobik incelemede toz örneklerinin %90,3’ü pozitif değerlendirilmiş ve 149 adet akar tespit edilmiştir. D. pteronyssinus %74, D. farinae %13, Dermatophagoides spp. gelişim formları %5, Cheyletus spp. %1, E. maynei %1, C. arcuatus %1, T. putrescentiae %1, L. destructor %1 ve tanımlanamayanlar %3 oranında belirlenmiştir. Der p 1 antijeni %93, Der f 1 antijeni ise %84,7 oranında tespit edilmiştir. Bir gram yatak tozunda saptanan en yüksek antijen miktarı Der p 1 için 1,272 μg, Der f 1 için ise 0,482 μg belirlenmiştir.
Çalışmanın yürütüldüğü illerde akar türleri ve dağılımları arasında fark gözlenmemiştir (p<0,05). Dermatophagoides spp. popülasyonunun %93’ünü oluşturmuştur. Depo/gıda akarlarının düşük (%4) oranda bulunmasının zeminlerden örnek alınmamış olmasıyla ilgili olduğu, ılıman ve nemli bölgelerde akarların aktivitesi yıl boyunca seyrettiği için yataklarda antijen birikiminin önemli olabileceği, antijen testlerinin akar tespitinde kullanılan mikroskobik yöntemlere ek olarak, akar alerjen yüklerinin ayrıntılı değerlendirilmesinde alternatif bir yöntem olarak kullanılabileceği ve duyarlı kişilerin yaşadığı ortamlar açısından bu teşhis yönteminin dikkate alınabileceği düşünülmektedir.

BACKGROUND: Allergen-specific immunoglobulins have a crucial role in allergic diseases. Most wheeze episodes develop before school age, and allergic rhinitis later develops during early elementary school years. However, the clinical background and cytokine/chemokine profiles associated with changes in immunoglobulins during early school-age are poorly understood.
METHODS: This study used blood samples from children participating in the JECS Pilot Study. We examined nineteen kinds of aeroallergen-specific immunoglobulins (IgE, IgG1, IgG4, and IgA) levels in patients at age 6 and age 8. Fluctuations of Der f 1- and Cry j 1-specific immunoglobulins levels during the two periods were compared to assess the frequency of allergic statuses and clusters of cytokine/chemokine profiles.
RESULTS: The medians of aeroallergen-specific IgE levels did not fluctuate, and almost all IgG1 and IgG4 decreased. In IgA, four (e.g., Der f 1) increased, whereas the other four (e.g., Cry j 1) decreased. The ratio of the Der f 1-specific IgG1 level at age 8 to that at age 6 was higher in children with poor asthma control than in children with better asthma control. Moreover, the cytokine/chemokine cluster with relatively lower IL-33 and higher CXCL7/NAP2 was associated with lower Der f 1- and Cry j 1-specific IgG4 levels, but not IgE levels.
CONCLUSIONS: The cluster of cytokine/chemokine profiles characterized by lower IL-33 and higher CXCL7/NAP2 was associated with the maintenance of aeroallergen-specific IgG4 levels. This result provides a basis for considering the control of aeroallergen-specific immunoglobulins.

A 12-year-old girl experienced anaphylactic shock and respiratory failure immediately after eating 4 pieces of takoyaki made at home. We suspected pancake syndrome because of the three reasons: First, the patient had a history of bronchial asthma and allergic rhinitis without any history of food allergies; Second, takoyaki flour that had been stored at room temperature for one month after opening the package had been used; and Third, both the specific IgE (ImmunoCAP® method) of Dermatophagoides farinae and Dermatophagoides pteronyssinus were found to be above the detection limit. Since the suspected flour had been discarded, a specimen of tako yaki was examined microscopically and 430 mites/g were identified. Although only 21.1ng/g of Der f 1 was detected by an ELISA, this amount was less than expected, based on the number of mites that had been observed. The decrease in the antigenicity of mite allergens due to heating and reducing agents, and the insolubilization of mite allergens due to disulfide bonds between the mite allergens and gluten were thought have made it difficult to identify mite antigens by ELISA.When pancake syndrome is suspected, and the flour that is thought to have caused the condition is difficult to obtain, microscopic identification of mites in cooked food should be considered.

As a method of evaluating the effect of inactivators on allergens while suppressing the effect of inactivator on the assay, we developed new dot-blot method that combines immunostaining and protein detection methods. This method is useful for evaluating whether the inactivator can inactivate allergens rather than removing them from the assay.

We have previously demonstrated that benzo(a)pyrene (BaP) co-exposure with dermatophagoides group 1 allergen (Der f 1) can potentiate Der f 1-induced airway inflammation. The underlying mechanism, however, remains undetermined. Here we investigated the molecular mechanisms underlying the potentiation of BaP exposure on Der f 1-induced airway inflammation in asthma. We found that BaP co-exposure potentiated Der f 1-induced TGFβ1 secretion and signaling activation in human bronchial epithelial cells (HBECs) and the airways of asthma mouse model. Moreover, BaP exposure alone or co-exposure with Der f 1-induced aryl hydrocarbon receptor (AhR) activity was determined by using an AhR-dioxin-responsive element reporter plasmid. The BaP and Der f 1 co-exposure-induced TGFβ1 expression and signaling activation were attenuated by either AhR antagonist CH223191 or AhR knockdown in HBECs. Furthermore, AhR knockdown led to the reduction of BaP and Der f 1 co-exposure-induced active RhoA. Inhibition of RhoA signaling with fasudil, a RhoA/ROCK inhibitor, suppressed BaP and Der f 1 co-exposure-induced TGFβ1 expression and signaling activation. This was further confirmed in HBECs expressing constitutively active RhoA (RhoA-L63) or dominant-negative RhoA (RhoA-N19). Luciferase reporter assays showed prominently increased promoter activities for the AhR binding sites in the promoter region of RhoA. Inhibition of RhoA suppressed BaP and Der f 1 co-exposure-induced airway hyper-responsiveness, Th2-associated airway inflammation, and TGFβ1 signaling activation in asthma. Our studies reveal a previously unidentified functional axis of AhR-RhoA in regulating TGFβ1 expression and signaling activation, representing a potential therapeutic target for allergic asthma.

UNASSIGNED: Even in subjects who are not sensitized to house dust mite (HDM), allergic symptoms can be clinically aggravated by exposure to dust. We previously reported that Dermatophagoides farinae (Df), an important HDM, or Der f 1, a major allergen of Df, induced the effector functions of eosinophils, which may be an important mechanism for HDM-induced symptoms in nonsensitized patients. In a clinical setting, β2-adrenergic agonists, such as salbutamol and formoterol, are used for the treatment of asthma attacks or exacerbation to release the airway obstruction. Several reports have suggested that some β2-adrenergic agonists have an anti-inflammatory capacity.
UNASSIGNED: In this study, we investigated whether β2-adrenergic agonist could modify the Df- or Der f 1-induced activation of eosinophils.
UNASSIGNED: Blood eosinophils obtained from healthy donors were preincubated with either formoterol (1 μM), salbutamol (1 μM), or buffer control and then stimulated with Df extract (1 μg/mL) or Der f 1 (100 pg/mL). Eosinophil adhesion to intercellular adhesion molecule (ICAM)-1 was measured using eosinophil peroxidase assays. Generation of superoxide anion (O2 -) was examined based on the superoxide dismutase-inhibitable reduction of cytochrome C. Eosinophil-derived neurotoxin (EDN) concentrations in cell media were measured by enzyme-linked immunosorbent assay.
UNASSIGNED: Formoterol, but not salbutamol, suppressed the Df- or Der f 1-induced eosinophil adhesion to ICAM-1. Furthermore, formoterol, but not salbutamol, suppressed Df-induced O2 - generation or EDN release. Neither formoterol nor salbutamol suppressed spontaneous eosinophil adhesion, O2 - generation, or EDN release.
UNASSIGNED: These findings suggested that formoterol, but not salbutamol, suppressed Df- or Der f 1-induced eosinophil activation when used at the same concentration. Therefore, formoterol could potentially be used for the treatment of bronchial asthma via both bronchodilation and anti-inflammatory effect.

OBJECTIVE: Bedding in childcare centers (CCCs) can hold house dust mite (HDM) allergens. This study examined whether HDM allergen levels can be reduced through the distribution of an educational newsletter on bedding control to parents of CCC children in Korea.
METHODS: All 38 CCCs were measured for Der 1 (sum of Der f 1 and Der p 1) concentrations on classroom floors and bedding before the intervention. Educational newsletters on children\'s bedding control were sent to 21 CCCs by mail, and teachers were asked to distribute the newsletters to the parents of the children (intervention group). The remaining 17 CCCs were not sent newsletters (control group). The measurement of Der 1 concentrations in 38 CCCs was repeated after the intervention. Dust samples were collected with a vacuum cleaner and analyzed using enzyme-linked immunosorbent assay methods.
RESULTS: The Der 1 concentrations on the bedding were significantly higher than those on the floors in 38 CCCs at baseline (p<0.05). Although changes of the Der 1 concentrations for the control group (n=17) were not significant, Der 1 concentrations for the intervention group (n=21) decreased significantly from 2077.9 ng/g dust to 963.5 ng/g dust on the floors and from 3683.9 ng/g dust to 610.4 ng/g dust on bedding (p<0.05).
CONCLUSIONS: The distribution of educational newsletters on bedding control to parents may be an effective means of controlling HDMs in CCCs.

Der f 1, a major house dust mite allergen and member of the papain-like cysteine protease family, can provoke immune responses with its proteolytic activity. To understand the role of Der f 1 in inflammatory immune responses, we studied the mechanism of the regulation of interleukin (IL)-8 expressions in human basophilic cell KU812 by proteolytically active recombinant Der f 1. Not only production of IL-8 mRNA was induced but also the DNA binding activity of activator protein-1 (AP-1) and phosphorylation of NF-κB p65 were increased in Der f 1-treated KU812. Furthermore, Der f 1 induction of IL-8 expression was sensitive to pharmacological inhibition of ERK and p38 mitogen activated protein kinase (MAPK) pathways. Der f 1 also activated ERK and p38 MAPK phosphorylation and rapidly induced reactive oxygen species (ROS) production. The antioxidant N-acetyl-cysteine (NAC) inhibited phosphorylation of ERK, but not p38, suggesting that secretion of IL-8 in KU812 cells treated with Der f 1 is dependent on ROS, ERK MAPK and p38 MAPK. We describe the mechanism of Der f 1-induced IL-8 secretion from human basophilic cells, which are thought to be important for allergic inflammation independent of IgE antibodies. These findings improve our understanding of the inflammatory immune response in human basophils to protease allergens.

BACKGROUND: Exposure to house dust mites (HDMs) is a major risk factor for the development of allergic symptoms. HDMs are worldwide in distribution. Assessing these allergens in each area is a critical step in evaluating the risk of sensitisation and controlling allergic symptoms. The aim of the present study was to evaluate the level of major HDMs, Der p 1 and Der f 1, in various parts of Iran.
METHODS: In 2009, 257 dust samples were obtained from living rooms\' carpets in seven cities throughout Iran with different geoclimatic conditions. The level of Der p 1 and Der f 1 was measured by commercial ELISA.
RESULTS: Detectable level of Der p 1 and Der f 1 levels were only found in Gorgan and Sari, two cities near the Caspian Sea with moderate temperatures and high relative humidity. In both of these cities, Der f 1 was more frequent than Der p 1 (100% vs. 85%) and was found to be at a higher level than Der p 1 (geometric mean 3128 vs. 439ng/g dust, P<0.001).
CONCLUSIONS: The results of this study confirm that house dust mites in Iran are restricted to the Caspian Sea coastal areas, and in other parts, due to seasonal variations of temperature and humidity mites are not able to grow well and therefore are not an important risk factor for sensitisation and respiratory allergies.