Morimitsu Tomikawa

Associations of functional NLRP3 polymorphisms with susceptibility to food-induced anaphylaxis and aspirin-induced asthma

BACKGROUND NLR family, pyrin domain containing 3 (NLRP3), controls the activity of inflammatory caspase-1 by forming inflammasomes, which leads to cleavage of the procytokines IL-1beta and IL-18. Recent studies have shown associations of human NLRP3 polymorphisms with susceptibility to various inflammatory diseases; however, the association with allergic diseases remains unclear. OBJECTIVE We sought to examine whether NLRP3 polymorphisms are associated with susceptibility to food allergy, food-induced anaphylaxis, and aspirin-induced asthma (AIA). METHODS We selected 15 tag single nucleotide polymorphisms (SNPs) of NLRP3 and conducted association analyses of NLRP3 using 574 and 1279 samples for food allergy and AIA, respectively. We further performed functional analyses of the susceptible SNPs. RESULTS Two NLRP3 SNPs (rs4612666 and rs10754558) were significantly associated with susceptibility to food-induced anaphylaxis (P = .00086 and P = .00068, respectively). The NLRP3 haplotype of the 2 SNPs also showed a significant association (P = .000098). We could confirm the association with susceptibility to another hypersensitivity phenotype, AIA (rs4612666, P = .0096). Functional analysis revealed that the risk alleles of rs4612666 and rs10754558 increased the enhancer activity of NLRP3 expression and NLRP3 mRNA stability, respectively. CONCLUSION Our results indicate that the NLRP3 SNPs might play an important role in the development of food-induced anaphylaxis and AIA in a gain-of-function manner. Further research on the NLRP3 inflammasome will contribute to the development of novel diagnostic and therapeutic methods for food-induced anaphylaxis and AIA.

Utility of the Peripheral Blood Basophil Histamine Release Test in the Diagnosis of Hen’s Egg, Cow’s Milk, and Wheat Allergy in Children

Background: The diagnosis of food allergy (FA) is made by oral food challenge tests (OFCs) that occasionally produce serious symptoms in patients; therefore, whether to perform OFCs should be carefully considered. The utility of the histamine release test (HRT) in the diagnosis of childhood FA has not been fully examined. Methods: Sixty-four subjects with suspected hen’s egg allergy, cow’s milk allergy (CMA), and wheat allergy (WA) were enrolled. The diagnosis of FA was made based on the outcomes of OFCs or a convincing history of symptoms after food ingestion within 6 months before or after sample collection. HRT was performed using an HRT Shionogi kit. The threshold of histamine release (HRT threshold), which was defined as the minimum concentration of food antigen to induce a 10% net histamine release, was analyzed in association with FA diagnosis. Results: Receiver operating characteristic analysis showed that the HRT threshold was useful in the diagnosis of heated egg allergy (HEA), raw egg allergy (REA), CMA, and WA. We were able to determine the cutoff value for the HRT threshold in relation to outcomes of OFCs. The cutoff value was 6 ng/ml of egg white antigen in HEA and REA (p < 0.01), 40 ng/ml of milk antigen in CMA (p < 0.01), and 500 ng/ml of wheat antigen in WA (p < 0.05). The efficiency was 70.3% for HEA, 78.0% for REA, 77.6% for CMA, and 70.7% for WA. Conclusions: We conclude that the HRT threshold measurement for egg white, milk, and wheat antigen is related to outcomes of OFCs and is useful in determining when OFCs should be performed.

Hypoproteinemia in severe childhood atopic dermatitis: A serious complication

As a complication of atopic dermatitis (AD), the incidence of hypoproteinemia is increasing among infants with severe AD in Japan. It can be a life‐threatening condition owing to hypovolemic shock as a result of hypoproteinemia and vascular infarction as a result of thrombocythemia. However, the pathophysiology of this condition remains unclear. The objectives of the present study were two‐fold. The first objective was to determine the main route of protein loss, i.e. through the damaged skin or the gastrointestinal tract, or as a result of insufficient food intake. The second objective was to identify whether allergy or infection was the cause of severe skin inflammation. Fifteen patients with AD were enrolled who had serum protein levels of 3.2–5.8 g/dl. Specific immunoglobulin E (IgE) and skin test to allergens, stool eosinophils, α1‐antitrypsin clearance, skin Staphylococcus aureus colonization and superantigens (SAgs) produced by the organism, serum SAg‐specific IgE antibodies, serum interleukin (IL)‐5, IL‐6, IL‐12, and interferon‐γ (IFN‐γ) were evaluated. Prominent serous skin discharge was seen in all of the patients and was found to have almost the same protein concentration as serum. Marked thrombocytosis, with a maximum of 1,060 × 103/ml, was seen. Skin culture revealed S. aureus colonization in all patients. SAg‐producing S. aureus were found in 84.6% of the patients. The concentration of serum IL‐5 was significantly increased and correlated well with the blood eosinophil count. Hence, the main route of protein loss was believed to be through damaged skin. The cause of severe inflammation was thought to be a combination of allergic inflammation and skin colonization by SAg‐producing S. aureus. Serum cytokines showed a T helper 2 (Th2) T‐cell‐mediated pattern. To prevent hypovolemic shock, vascular occlusion, and growth retardation, it is of vital importance to diagnose hypoproteinemia at an early stage and start appropriate therapy.

Dolphin-Assisted Seawater Therapy for Severe Atopic Dermatitis: An Immunological and Psychological Study

We performed seawater therapy on atopic dermatitis patients for 6 days with dolphins. The presence of the dolphins seemed to alleviate the pain patients experienced while bathing in seawater. The skin condition improved dramatically, and immunologically, serum IL-8 levels decreased but MIP-1α and eosinophil cationic protein did not change.

Human mast cell progenitors in peripheral blood from atopic subjects with high IgE levels.

Background It remains unclear whether the number of circulating mast cell progenitors is increased in patients with atopic diseases. Distinct genotypes are reported to affect mast cell/basophil activation.

Human Mast Cell Colony-Forming Cells in Culture

Human mast cells require an extra long period for their development from their progenitors as compared with other cell types. We have recently established the method of forming human mast cell colonies by culturing purified cord blood cells and peripheral blood cells without in vivo priming of cytokines in serum-free methylcellulose supplemented with stem cell factor (SCF) and cytokines, and by laying the fresh methylcellulose medium every 10–14 days [1]. The majority of mast cell progenitors were multipotent, i.e. capable of producing other cell types such as erythorid cells. However, they became colonies consisting of pure mast cells after 6 weeks of culture. The methylcellulose system was necessary to obtain a large enough number of mast cells especially when semipurified hemopoietic cells were cultured. We could retrieve almost pure mast cells after 6 weeks by dissolving methylcellulose and could culture the cells for at least several months in liquid medium. Here, we discuss the colony formation of human mast cells in methylcellulose mainly by describing the methodology in detail. Methodology

The Skin Prick Test is Not Useful in the Diagnosis of the Immediate Type Food Allergy Tolerance Acquisition

BACKGROUND Some studies have been reported about positioning of SPT in the diagnosis of food allergy. On the other hand, it is not yet clear about the positioning of SPT in the diagnosis of tolerance acquisition of the immediate type food allergy. METHODS The retrospective study had been conducted for 236 egg allergic children (51.3 months in mean), 127 milk allergic children (53.4 months), and 96 wheat allergic children (42.6 months). The retrospective analysis of serum nonspecific and antigen-specific IgE levels, SPT, and OFC had been conducted for each allergic patient. All OFC had been conducted to verify the acquisition of tolerance against eliminated food. RESULTS The OFC was positive in 61 (25.8%) hens egg allergies, 34 (26.8%) milk allergies and 33 (34.4%) wheat allergies. The greatest AUC for each allergen is as following; hens egg for egg white specific IgE at 0.745, milk/histamine (wheal) index at 0.718, and wheat for wheal size at 0.597. For the predictive decision points, the highest accuracy rate was at 25.8% for an egg white wheal of 9.5mm, at 26.8% for a milk wheal of 9.5mm, and at 34.4% for a wheal of 6.5mm. CONCLUSIONS As a result of this analysis, the diagnostic accuracy of SPT had not been satisfactory to judge the acquisition of tolerance in allergic children for eggs, milk and wheat. Therefore, this is not a strong evidence to testify the tolerance of the immediate type food allergy.

Asthma diagnosis and treatment – 1005. Optimization for the withdrawal of inhaled corticosteroid treatment by monitoring fractional exhaled nitric oxide (feno) and lung functions

Results Subjects in recurrent asthma symptom group were 28 cases and those of non-recurrent asthma symptom group 27 cases (relapse rate: 50.9%). Any significant factors in background patients’ profiles, such as FeNO and pulmonary functions, were not associated with the recurrence of asthma. In recurrent asthma symptom group, FeNO was significantly increased by 3 months after withdrawal of ICS (from 31.8 ppb to 49.2 ppb). Among recurrent asthma symptom group, pulmonary functions were significantly decreased within 1 months (FVC: from 2.11L to 2.02L, FEV1.0: from 1.93L to 1.85L and %FEV1.0: from 98.1% of to 93.8%). Conclusions Although these factors at the time of ICS withdrawal could not predict asthmatic revival, it is highly recommended to follow asthmatic patients who quit ICS therapy by measuring pulmonary function and FeNO periodically.

Budesonide inhalation suspension versus methylprednisolone for treatment of moderate bronchial asthma attacks

BackgroundOwing to their side effects, administration of steroids for bronchial asthma attacks should be minimized. We investigated whether budesonide inhalation suspension (BIS) could replace intravenous steroid administration for the treatment of moderate bronchial asthma attacks.Subjects and MethodsThe subjects were children aged 5 years and younger hospitalized for moderate bronchial asthma attacks. Patients were randomly assigned to one of two groups: 20 patients received methylprednisolone (mPSL) and 20 were treated with BIS. The mPSL group began treatment with inhalation of procaterol hydrochloride (0.3 mL) and disodium cromoglycate (2 mL) three times a day and systemic administration of mPSL (1 mg/kg) three times a day. The BIS group began treatment with inhalation of procaterol hydrochloride (0.3 mL) and BIS (0.5 mg) three times a day. The frequency of inhalations and steroid administration was adjusted according to the severity of symptoms. The cortisol level at discharge was measured.ResultsThere were no significant differences between the two groups in terms of the severity of attacks and duration of management, or in terms of therapeutic efficacy, duration of wheezing, or period of hospitalization. The frequency of inhalations on days 3 to 6 of hospitalization was lower in the BIS group than in the mPSL group, and the cortisol level at discharge was significantly higher in the BIS group (13.9 ± 6.1 μg/dL) than in the mPSL group (8.0 ± 2.1 μg/dL) (p = 0.008).ConclusionIn patients with recurrent wheezing or bronchial asthma of <5 years, the efficacy of BIS is equivalent or better than mPSL for moderate bronchial asthma attacks, and in contrast to steroid treatment, BIS treatment do not suppress adrenocortical function.

Food allergy and anaphylaxis – 2054. Easy-to-use severity grading system for treatment of symptoms induced by oral food challenge

Methods From June 2008 to June 2012, the severity of SR was assessed at double-blind placebo-controlled food challenge test (DBPCFC) to evaluate if they were candidates for rush oral immunotherapy or not. The medical records of 342 patients who showed positive reaction at DBPCFC were analyzed. A hundred and forty-one were allergic to hen’s egg, 156 to milk and 45 to wheat. We modified the grading system proposed by Sampson HA in 2003 to enhance the convenience at clinical practice. It was proposed to indicate “severity of SR for each organ system, i.e., skin, mucosa, gastrointestinal tract, respiratory tract, cardiovascular, and neurological system. Systemic reactions for each organ were classified as Grade (G) 1 (mild), G2 (moderate), and G3 (severe). The severity score was based on the organ system mostly affected. We examined relationship between the severity score and its treatment during DBPCFC.