Allergic patients mount a Th2 response to common allergens, like house dust mite (HDM), pollens, molds and animal dander. Most inhaled antigens are immunologically inert, however if these antigens are accompanied by microbial or endogenous danger patterns (alarmins), they can be recognized by inflammatory cells. Dendritic cells are the most potent antigen presenting cells, which express a wide variety of receptors on their cell surface, recognizing these microbial patterns, damage induced molecules and cytokines. Dendritic cells become reporters of the microenvironment if exposed to the allergen, subsequently migrating to the draining lymph nodes where they activate naïve T lymphocytes. Dendritic cells could also be indirectly activated by epithelial cells, which express various receptors and secrete a variety of cytokines early after allergen exposure. Upon HDM exposure these cells secrete chemokines to attract monocytes and immature dendritic cells, and GM-CSF, TSLP and IL-33 to activate dendritic cells, mast cells and basophils. Danger signals which alert dendritic cells and epithelial cells comprise many proteins and molecules, contributing to an enhanced immune response to inhaled allergens. This review focuses on the role of dendritic cells and alarmins in the sensitization to inhaled allergens in allergic asthma.

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Urticarial rash, one of the clinical manifestations characteristic of cryopyrin-associated periodic syndrome (CAPS), is caused by a mutation in the gene encoding for NLRP3 (nucleotide-binding oligomerization domain, leucine-rich repeats containing family, pyrin domain containing 3). This intracellular pattern recognition receptor and its adaptor protein, called apoptosis associated speck-like protein containing a caspase-recruitment and activating domain (ASC), participate in the formation of a multi-protein complex termed the inflammasome. The inflammasome is responsible for activating caspase-1 in response to microbial and endogenous stimuli. From the analysis of cellular mechanisms of urticarial rash in CAPS, we have traced caspase-1 activated IL-1β in CAPS to a surprising source: mast cells. Recently, two groups have generated gene-targeted mice that harbored Nlrp3 mutations. These mice had very severe phenotypes, with delayed growth and the development of dermatitis, but not urticaria. The reason for the differences in the skin manifestations observed with CAPS and these knock-in mice relates to the findings that the inflammasome also plays a role in contact hypersensitivity, and that IL-18, another cytokine involved with inflammasome-activation of caspase-1, may be a major player in dermatitis development.

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Hemozoin, a bio-crystalline substance, is a hemin detoxification by-product of malaria parasites. The role of hemozoin crystals in host immune system modulation by malaria parasites, and how they interact with the immune system has been enigmatic. Here, we summarize recent progress in our understanding of how hemozoin might be interacting with the host immune system. In particular, the potential application of hemozoin crystals as an adjuvant may provide insights into the molecular mechanisms involved in immune responses to malarial infection and provide a rationale for the design of vaccines against malaria as well as other immunological disorders such as allergies.

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IL-1 is a well-characterized proinflammatory cytokine that is involved in host defense and autoimmune diseases. IL-1 can promote activation of T cells, including Th1 cells, Th2 cells and Th17 cells, and B cells, suggesting that IL-1 may contribute to the development of various types of T-cell-mediated diseases. This report reviews and discusses the role of IL-1 in the pathogenesis of allergic diseases based on studies using IL-1-related gene-deficient mice.

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Th1 cells, which express IL-18R, produce IFN-γ in response to Ag and IL-2 and increase further production of IFN-γ upon additional IL-18 stimulation. They simultaneously produce Th2 cytokines (IL-9 and IL-13), GM-CSF and chemokines (RANTES, MIP-1α). Human Th1 cells also produce IFN-γ and IL-13 in response to anti-CD3 and IL-18. Recently, we demonstrated Th1 cells induce intrinsic type atopic asthma and dermatitis by production of Th1- and Th2-cytokines and chemokines. Here, we review the pathological roles of Th1 cells, stimulated with Ag and IL-18 in vivo, in the pathogenesis of allergic disorders by production of Th1 and Th2 cytokines and chemokines. Based on this unique function of Ag- plus IL-18-stimulated Th1 cells, we proposed to designate them as "super Th1 cells".

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Interleukin-33 (IL-33) is a member of the IL-1 cytokine family, which includes IL-1 and IL-18. IL-33 is considered to be crucial for induction of Th2-type cytokine-associated immune responses such as host defense against nematodes and allergic diseases by inducing production of such Th2-type cytokines as IL-5 and IL-13 by Th2 cells, mast cells, basophils and eosinophils. In addition, IL-33 is involved in the induction of non-Th2-type acute and chronic inflammation as a proinflammatory cytokine, similar to IL-1 and IL-18. In this review, we summarize and discuss the current knowledge regarding the roles of IL-33 and IL-33 receptors in host defense and disease development.

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Background: Th17-inducing activity is carried by certain polysaccharides such as β-glucan derived from Candia albicans. Our previous studies have shown that Th1- and Th2-inducing activities can be qualitatively evaluated by the expression patterns of Notch ligand isoforms, using human monocyte-derived dendritic cells (Mo-DCs) and some leukemic cell lines such as THP-1. The association of Th17-inducing activities with Notch ligand expression patterns has been unclear.
Methods: Mo-DCs from healthy volunteers were co-cultured with HLA-DR-nonshared allogeneic CD4⁺ naïve T cells to induce a mixed lymphocyte reaction, in the presence of adjuvants, such as curdlan. Culture supernatants were assayed for IFNγ, IL-5 and IL-17 by an enzyme-linked immunosorbent assay (ELISA). Notch ligand expression on Mo-DCs and THP-1 cells was evaluated by using RT-PCR.
Results: The present study shows that curdlan, one of the β-glucans, has the ability to induce DC-mediated Th17 differentiation. It is also interesting to note that Jagged1 mRNA in Mo-DCs and THP-1 cells is up-regulated by curdlan. Furthermore, polyclonal anti-Jagged1 antibody inhibited such DC-mediated Th17 differentiation.
Conclusions: This study suggests that curdlan induces human DC-mediated Th17 polarization via Jagged1 activation in DCs.

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Background: We have previously demonstrated that addition of omalizumab to standard therapy improved asthma control by significantly improving lung function and reducing asthma exacerbations in Japanese patients with moderate-to-severe asthma. The aim of this study was to evaluate the effects of omalizumab on long-term disease control in Japanese patients with moderate-to-severe persistent asthma.
Methods: An open-label, 48-week study was conducted in 133 Japanese patients with moderate-to-severe persistent asthma. Omalizumab was administered subcutaneously every 2 or 4 weeks based on serum IgE level and body weight in each patient.
Results: Treatment with omalizumab significantly improved lung function. A subgroup of patients with inadequately controlled severe persistent asthma, despite high dose inhaled corticosteroids and other multiple controller therapies, which corresponds to the Japanese label (label population), showed greater improvements in morning PEF and FEV₁ than the whole study population (full Analysis Set). Serum free IgE levels decreased to below the target and were maintained during the treatment period in almost all patients. The majority of adverse events were mild-to-moderate in severity and there was no trend toward an increase in incidence of adverse events with increase in duration of omalizumab. In addition, the profile of adverse events in this study was similar to that in a 16-week, placebo-controlled study which the present authors had conducted previously in Japan. There were no anaphylactic reactions and no anti-omalizumab antibodies were detected.
Conclusions: Long-term treatment with omalizumab is effective and well tolerated in Japanese patients with moderate-to-severe persistent asthma.

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Background: Ovalbumin, ovomucoid, ovotransferrin, lysozyme, and ovomucin are known to be major allergens found in egg white. Egg white protein is composed of over 30 proteins; many of which have neither been identified nor their allergenicities characterized. This study set out to analyze whether unknown proteins that bind to IgE antibodies in serum from patients with egg allergy exist in egg white.
Methods: Diluted egg white proteins were separated by 2-dimensional (2-D) gel electrophoresis. Immunolabeling was performed on individual patient sera from 19 child patients with egg white allergy and 11 negative control subjects. Spots of egg white proteins that bound to the patients' IgE were identified by mass spectrometry-based proteomics.
Results: Egg white proteins were separated into 63 spots. Twenty-five of the 63 reacted with egg allergy patients' sera, and 10 of the 25 reactive spots showed IgE-reactivity to controls as well. Specific bindings to the IgE from egg allergy patients were found in 15 spots; one of which was confirmed as ovotransferrin. Among the other 14 protein spots, egg white cystatin and lipocalin-type prostaglandin D synthase (L-PGDS) were newly identified proteins that reacted with IgE in patients with egg allergy.
Conclusions: We demonstrated that L-PGDS and cystatin reacted with serum IgE in patients with egg allergy. Our proteomics-based analysis in egg white gives a comprehensive map of proteins bound with IgE and should assist in enabling more accurate diagnoses and recommendations of desensitizing treatments for individual patients.

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Background: Chronic cough is the only symptom of cough variant asthma (CVA) and atopic cough (AC). Cysteinyl leukotriene receptor antagonists have been shown to be effective in CVA, but there are no reports on their effectiveness in AC. To evaluate the antitussive effect of montelukast, a leukotriene receptor antagonist, in CVA and AC.
Methods: Seventy-five patients with chronic cough received diagnostic bronchodilator therapy with oral clenbuterol hydrochloride for 6 days. Of the 75 patients, 48 and 27 met the simplified diagnostic criteria for CVA and AC, respectively. Patients with CVA were randomly divided into 3 groups: montelukast, clenbuterol, and montelukast plus clenbuterol. Patients with AC were randomly divided into 2 groups: montelukast and placebo. The efficacy of cough treatment was assessed with a subjective cough symptom scale (0 meant "no cough" and 10 denoted "cough as bad as at first visit"). The cough scale, pulmonary function test, and peak expiratory flow rate (PEF) were evaluated before and after 2 weeks of treatment.
Results: In patients with CVA, 2-week treatment with montelukast, clenbuterol, and montelukast plus clenbuterol all significantly decreased cough scores and treatment with montelukast plus clenbuterol was superior to treatment with montelukast alone. In the montelukast plus clenbuterol group, PEF values in the morning and evening significantly increased after 2 weeks compared with values before treatment. In patients with AC, scores on the cough scale did not differ significantly between the montelukast group and the placebo group.
Conclusions: Montelukast was confirmed to suppress chronic non-productive cough in CVA, whereas it was not effective in non-productive cough in AC.

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Background: The ability to predict the development of allergic diseases in infants is important. Predictive biomarkers are wanted to improve the risk evaluation in addition to known heredity of allergy. Biomarkers taken during infancy need to be evaluated through longitudinal studies into adulthood. The objective of this study was to analyse the occurrence of metachromatic cells in the nasal mucosa during infancy (MC_infancy) and evaluate the cells as predictive biomarkers of allergy development.
Methods: Previously, MC_infancy occurrences were analysed in 64 infants with and without allergy heredity, and related to allergy development at 18 months and 6 years of age. In this third follow-up at 18 years of age, current allergy symptoms were analysed. MC_infancy findings were related to the cumulative number of allergic subjects. The predictive values of MC_infancy and known heredity were compared.
Results: The cumulative number of subjects with allergy was 46, probable allergy 5, and no allergy 13. Detected MC_infancy predicted allergy with high accuracy (31/33), but negative MC_infancy findings did not exclude the risk (15/31). In the group of allergic subjects positive MC_infancy were found in 31/46 (67%), positive heredity in 37/46 (80%) and one/both factors positive in 43/46 (93%). Detection of MC_infancy could precede the debut of allergy symptoms by many years.
Conclusions: Detected MC_infancy predicted allergy development, but absence of MC_infancy did not exclude the risk, and therefore this biomarker was not found to be adequate. There is a further need to find biomarkers with high ability to both predict and exclude the risk.

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Background: To clarify the mechanism of stress-induced modification of allergic diseases, we studied the effect of restraint stress on plasma levels of cytokines and the symptoms of pollinosis in mice.
Methods: The effects of restraint stress and the role of the hypothalamo-pituitary-adrenal axis (HPA-axis) in the development of pollen antigen-induced pollinosis were studied in control, hypophysectomized, adrenalectomized or ACTH-administered mice. Twenty days after sensitization, animals were subjected to mild restraint stress for 3 hours, and plasma levels of IFN-γ, IL-10, and IgE were measured. We analyzed the incidence of sneezing and nasal rubbing in the sensitized animals.
Results: Plasma levels of IL-10 and IgE increased in the sensitized animals with a concomitant increase in the incidence of sneezing and nasal rubbing. The increases in plasma IgE, IL-10 and the incidence of sneezing and nasal rubbing were suppressed by restraint stress. Adrenalectomy increased IFN-γ, inhibited the increase in plasma IL-10 and IgE, and suppressed the incidence of sneezing. In contrast, hypophysectomy increased plasma levels of IL-10, IFN-γ, and IgE and the incidence of sneezing. Intraperitoneal administration of ACTH decreased IL-10 in plasma but increased IFN-γ and suppressed the incidence of nasal rubbing.
Conclusions: The present findings show that the HPA-axis and ACTH play important roles in the regulation of plasma cytokines and IgE thereby modulating symptoms of pollinosis. The results also suggest that a mild restraint stress suppresses the increase in Th2-dependent cytokines and IgE to reduce the symptoms of pollinosis.

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Background: The number of amphiregulin (AR)-positive mast cells in the bronchial mucosa and the levels of AR in sputum from asthmatic patients have been reported to be increased. In addition, AR can promote mucin gene expression in human epithelial cells, suggesting that AR contributes to the pathogenesis of allergic asthma.
Methods: To elucidate the role of AR in the pathogenesis of asthma, we immunized AR-deficient mice with ovalbumin (OVA) and then induced airway inflammation in them after OVA inhalation. The OVA-induced airway inflammation was assessed on the basis of the lung histology, number of leukocytes in the bronchoalveolar lavage (BAL) fluid, Th2 cytokine levels in the BAL fluid and OVA-specific IgG1 and IgE levels in the serum and compared between AR-sufficient and -deficient mice.
Results: The OVA-induced airway inflammation was comparable in the AR-sufficient and -deficient mice.
Conclusions: Amphiregulin is not essential for induction of acute airway inflammation by OVA in mice.

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Background: The inhalation of Salsola kali pollen is an important cause of pollinosis during summer and early fall throughout desert and semi-desert areas. Sal k 1 has been previously reported as a major allergen of S. kali pollen. In this study, we produced the recombinant Sal k 1 and also its low IgE-binding mutant form. We further compared the IgE binding ability of these two recombinant molecules.
Methods: The recombinant Sal k 1 and its low IgE-binding variant, obtained by three amino acid exchanges (R₁₄₂→S, P₁₄₃→A, D₁₄₄→V), were cloned and expressed in E. coli, as proteins fused with thioredoxin and His-tags, and then purified by Ni²⁺ affinity chromatography. The IgE-binding capacity of the wild-type and mutated rSal k 1 was compared using immunoblotting, ELISA and inhibition assays by ten sera from S. kali allergic patients. Moreover, in vivo IgE-reactivity was investigated by the skin prick test.
Results: Both the recombinant and the mutated form of Sal k 1 were expressed in E. coli at a relatively high amount and soluble form. All sera recognized rSal k 1 via immunoassay analysis. In addition, inhibition assays demonstrated that the purified rSal k 1 was similar to its counterpart in the crude extract. The mutated rSal k 1 exhibited a reduced IgE-binding capacity against wild-type rSal k 1.
Conclusions: This study demonstrates that purified rSal k 1 is comprised of IgE-epitopes similar to that of its natural counterpart and that the mutated variant showed a reduced IgE-binding capacity based on in vitro assays and in vivo provocation testing.

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Background: In the management of hypersensitivity pneumonitis (HP), antigen avoidance is crucial to prevent the progression of disease. Indirect and unrecognized exposure to the antigen may continue for a long time if persistence of the causative antigen is not recognized. To make a correct assessment of the patients' environment, we tried to establish the methods to detect indoor and outdoor avian antigens.
Methods: Sixteen patients with bird-related HP, 4 asymptomatic breeders, and 6 healthy controls were examined. We prepared anti-pigeon dropping extracts (PDE) polyclonal antibody from rabbits. Air samples and house dust samples were analyzed by an antigen-capture ELISA with signal amplification using catalyzed reporter deposition.
Results: In air samples, avian antigen could be detected in patients with HP (0.73 ± 0.53ng/m³) and asymptomatic breeders (0.63 ± 0.23ng/m³). In house dust samples, the amount of avian antigen was higher in patients with HP (2.4 ± 1.8μg/g) and asymptomatic breeders (4.1 ± 2.3μg/g) than in the controls (0.1 ± 0.2μg/g).
Conclusions: Detection of indoor and outdoor avian antigen might contribute to the correct diagnosis and appropriate managements of bird-related HP.

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