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Background and Methods: The objective of this study was to analyze the sensitization to casein and β-lactoglobulin (BLG) in children with cow's milk allergy (CMA) in Japan. To this end, 115 CMA children were selected on the basis of the presence of cow's milk-specific IgE antibodies in serum and compatible clinical history. Specific IgE antibodies against casein and BLG were determined using CAP-RAST (considered positive when score 2 or more). Results: Titer of anti-casein IgE was significantly higher than that of anti-BLG IgE in CMA patients. IgE antibodies specific to casein were positive in 107 patients (97.3%), while those to BLG were positive in 51 patients (46.6%). Forty-eight patients (43.6%) were positive to both casein and BLG. We divided patients to two groups who were sensitized to casein only (C group) and who were sensitized to both casein and BLG (C/B group). No significant difference was seen in sensitization rate to white egg between C/B group and C group. However titer of anti-white egg IgE was significantly higher in C/B group than C group. As for sensitization rate and levels of specific antibodies to mite and Japanese cedar pollen there was no difference between two groups. Rates of resolution of CMA at the 3 years of age were higher in the C group than C/B group. Conclusion: In conclusion we found that casein is a major allergen of cow's milk allergy in Japanese children. Patients who are sensitized to several milk allergens are likely to be more sensitized to other food allergens. Sensitization to several milk allergens tends to have poor prognosis of CMA. f © 2010 Japanese Society of Allergology.

Background: House dust extract is used in conventional immunotherapy for house dust-mite (HDM) allergic rhinitis in Japan. However, an alternative administration route is desired. The aims of the present double blind, placebo-controlled trial were to evaluate the therapeutic efficacy and safety of sublingual immunotherapy (SLIT) with house dust extract in pediatric patients with HDM allergic rhinitis. Methods: The study population comprised 31 subjects (21 males and 10 females) aged from 7 to 15 years old. Twenty patients (the active group) received house dust extract and 11 received placebo via sublingual administration. Extract or placebo (1 ml) was administered at 10-fold dilution once weekly for 40 weeks. During the study period, the subjects recorded their daily nasal symptoms and use (dose and frequency) of other medications in a nasal allergy diary. Results: The symptom scores in the active group began to decrease about 24 weeks after initiation of treatment and significant differences between the active and placebo groups were observed after 30 weeks. The average scores for the last four weeks of the study were significantly lower than those for the first four weeks in the active group but not in the placebo group. The only local adverse effect was a bitter taste reported by one patient. There were no other local or systemic adverse effects associated with SLIT. Conclusions: Our results suggest that SLIT with house dust extract for more than 30 weeks is safe and effective treatment for HDM allergic rhinitis in children.© Japanese Society of Allergology.

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-1 beta 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. (J Allergy Clin Immunol 2009;124:779-85.)

The regulatory IL-10 and TGF-beta 1 cytokine gene polymorphisms have been associated with allergic diseases in different populations, like Caucasian, Chinese and Indians. We investigated the association between the polymorphisms IL-10 A-1082G, C-819T, C-627A and TGF-beta 1 T+869C, G+915C, C-509T and food allergy in Japanese children. One hundred and eleven children with food allergy and 115 atopic control children without food allergy were recruited. DNA samples from these subjects were genotyped by using PCR. The odds ratio of IL-10 -1082 AA genotype was 2.5 (95% CI, 1.0-6.4) for food allergy risk when compared with atopic control subjects (p = 0.03). There were no significative differences in the frequency of TGF-beta 1 gene polymorphisms between both groups. Our results indicate that IL-10 A-1082G gene polymorphism is associated with food allergy susceptibility in atopic Japanese children.

Our objective was to investigate the efficacy of intravenous alendronate for the treatment of glucocorticoid-induced osteoporosis (GIOP) in children with autoimmune diseases. Five children with autoimmune disease and GIOP were treated with 5 mg intravenous alendronate once every 3 months. After 1 and 2 years, we evaluated the changes in the Z score of the femoral neck bone mineral density (BMD), serum bone alkaline phosphatase, and urinary deoxypyridinoline. Six patients with GIOP, whose BMD could be observed over a 1-year period without alendronate treatment, were defined as controls. After 1 and 2 years of treatment, intravenous treatment significantly inhibited bone loss. The efficacy of alendronate demonstrated a significant correlation with a high level of bone turnover markers before alendronate treatment. Intravenous alendronate is considered to be a good choice for the treatment of GIOP because of its excellent efficacy. In addition, our study suggests that the efficacy of alendronate depends on the bone turnover of patients before treatment. Intervention with bisphosphonates during periods of high bone turnover may be recommended.

Recent studies have suggested that intestinal microbiota play a substantial role in the development of allergic diseases during infancy. We analyzed fecal microbiota in 18 Japanese infants with or without allergy at 6 months and 2 years of age using a cell culture technique. Allergy determination was based on doctor-diagnosed allergic diseases and skin prick tests. There were no differences between 9 allergic and 9 non-allergic infants at 6 months of age in the frequencies or counts of 13 genera and yeast-like organisms. Bifidobacterium was dominant in all infants irrespective of allergy status. At 2 years of age, 8 infants were non-allergic and 10 infants were allergic. Allergic infants at 2 years of age had higher counts of Bacteroides and higher ratios of Bacteroides to Bifidobacterium than non-allergic infants. Despite the small population size used in this study, the results support a significant role of Bacteroides in the pathogenesis of allergy during infancy.

We previously showed that immunization of mice with murine fibroblasts transfected with the thyrotropin receptor (TSHR) and a murine major histocompatibitity complex (MHC) class II molecule induces immune thyroid disease with the humoral and histological features of human Graves' disease in about 20% of mice. In this model, based on the proliferative response of Tcells from hyperthyroid mice to a panel of overlapping TSHR peptides, we now demonstrate that TSHR 121-140 peptide contains an immunodominant Tcell epitope. Supporting this conclusion, spleen cells from mice immunized with TSHR 121-140 peptide showed a strong proliferative response to fibroblasts transfected with the TSHR and a murine I-A(k) molecule, but not either atone. Also, intranasat administration of 100 mu g of TSHR 121-140 peptide led to suppressed proliferative response of Lymph node cells to the peptide. Interestingly, however, administration of this peptide enhanced, rather than suppressed, the frequency and severity of Graves' disease induced by the immunization of the fibroblasts transfected with the TSHR and a murine I-A(k) molecule. Spleen cells from hyperthyroid mice that were pretreated with intranasal peptide tended to produce lesser amounts of IL-4, IL-10 and IFN-gamma than those from normothyroid control mice. Although precise mechanisms of this enhancement remain to be determined, the results suggest that attempts to treat Graves' disease by intranasat administration of an immunodominant TSHR Tcell epitope may aggravate, not prevent, the disease. (C) 2007 Elsevier Inc. ALL rights reserved.

Background: Respiratory syncytial virus (RSV) G protein is involved in Th2-shifted immune response, while F protein has a reverse effect on RSV infection in Th2-prone BALB/c mice. Studies on the human T cell response to F or G protein are few, and the relationship between the immune response to G protein and atopy is not known. Methods: We established CD4+ RSV-specific T cell lines (TCLs) from adult patients with respiratory allergic diseases (allergics) or nonatopic controls (controls), and examined proliferative responses and gamma-interferon (IFN-gamma)and interleukin 4 (IL-4) production in these TCLs upon stimulation with RSV, F or G proteins. Results: 32 and 29 RSV-specific oligoclonal TCLs were established from allergics and controls, respectively. IL-4/IFN-gamma in the culture supernatant of antigen-stimulated TCLs was significantly higher in allergics than in controls (p = 0.042). IL-4/IFN-gamma ratios in the culture supernatants of G-protein-reactive TCLs were significantly higher in allergics than in controls (p = 0.016), while no differences in IL-4/IFN-gamma in culture supernatants of F-protein-reactive TCLs were found between allergics and controls (p = 0.787). IL-4/IFN-gamma in the culture supernatants of G-protein-reactive TCLs was significantly higher than those of F-protein-reactive TCLs in allergics (p = 0.023) but not in controls (p = 0.768). Conclusion: The results suggest that the T cell response to RSV is influenced by the atopic diathesis as well as by individual RSV antigens, and that G protein may be an important antigen involved in allergy in humans. Copyright (c) 2008 S. Karger AG, Basel.

Cogan's syndrome is a rare inflammatory disease characterized by nonsyphilitic ocular interstitial keratitis associated with hearing loss and vestibular impairment. Although systemic corticosteroids are usually used as the standard therapy, hearing ability in most cases gradually deteriorates. We, herein, describe a patient with childhood Cogan's syndrome who was treated with low-dose oral methotrexate, which successfully helped to taper the doses of the systemic corticosteroids. The serum levels of the complements were good markers for the disease activity in this patient.

Background. The contribution that genetic polymorphisms of Toll- like receptor ( TLR) 4 and of CD14 - both of which recognize respiratory syncytial virus ( RSV) in the innate immune response - make to RSV bronchiolitis in the Japanese population has not yet been clarified.Methods. This study genotyped 2 TLR4 mutations, Asp299Gly and Thr399Ile, and 2 single- nucleotide polymorphisms (SNPs) of CD14, -550 C/T and -159 C/T, in 54 children with RSV bronchiolitis and in 203 control subjects. CD14 SNPs and the serum level of soluble CD14 (sCD14) also were examined in 67 cord- blood specimens and in serum samples from 73 6-year-old children.Results. No TLR4 mutations were found. The frequencies of both the CC genotype and the C allele of CD14 -550 C/T were significantly higher in children with RSV bronchiolitis than in the control subjects. The serum level of sCD14 was significantly higher in children with the CC genotype of CD14 - 550 C/ T than in those with the CT and TT genotypes.Conclusions. CD14 -550 C/T, which is related to the serum level of sCD14, is associated with the development of RSV bronchiolitis in the Japanese population. This study's results indicate that, in different ethnic groups, different genetic factors contribute to the development of RSV bronchiolitis.

Background: Clarification of the mechanisms underlying the development of food-sensitive intestinal inflammation will provide an important clue to combating food allergies. Objective: To establish a model of intestinal inflammation caused by oral administration of antigen without additional treatments, we focused on the ovalbumin (OVA) 23-3 T-cell receptor transgenic mouse, which had been reported to have high serum antigen-specific IgE responses to the feeding of an egg white diet. Methods: Changes in body weight of mice fed an egg white diet were monitored throughout tire 28-day experimental period. After the 28-day feeding, intestinal tissues were harvested for histologic examination. Endogenous production of cytokines and histamine in the jejunum, and production of cytokines secreted by OVA-specific CD4(+) T cells purified from mesenteric lymph nodes, were analyzed. Results: Egg white diet-fed OVA23-3 mice developed weight loss and inflammation with villous atrophy and goblet cell hyperplasia, especially in the jejunum. A further characteristic feature was evidence of weight recovery and tissue repair. Jejunal inflammation was also observed in egg white diet-fed recombination activating gene (RAG)-2-deficient OVA23-3 mice. In addition, tissue sections revealed significant infiltration of specific IgE-positive cells and IgE-positive degranulating mast cells. Higher levels of IL-4 and significant levels of histamine were detected in the tissues. In the supernatant of OVA-stimulated T cells, IL-10 levels were also markedly elevated. Conclusion: We report that high-dose and continuous intake of primitive OVA alone induces enteropathy containing regions under repair in OVA23-3 mice. Antigen-specific T cells and inflammatory cells primed by T(H)2- responses play important roles in regulation of development and improvement of the disease. Clinical implications: Long-term antigen intake causes T(H)2-dependent and food-sensitive enteropathy followed by tissue repair.

The combination of the local disinfection therapy against Staphylococcus aureus with the conventional therapy for atopic dermatitis has been widely used, and the improvement in skin lesions has been reported to be associated with a remarkable decrease in IgE levels and reagin antibody titers. We have already reported that affected organs were not only the skin but also the gastrointestinal tract in a case with atopic dermatitis. In the present study, the duodenal tissues were examined by biopsy in 32 patients with atopic dermatitis, and mild or chronic duodenitis was observed in all samples. Toxins we re examined by PCR from 180 Staphylococcus aureus strains obtained from our patients. The detection rate of toxins was 82.8%. In many patients, antitoxin IgE antibody titers corresponding to their types of toxin and IgE levels were decreased in a parallel manner as time passed. We found 1 patient who complained of paresthesia in all four limbs, and her neurological and radiological examinations showed moderate cervical spondylosis. Neurological examinations revealed some abnormalities in 43 out of 50 patients with atopic dermatitis, such as hyperreflexia of the legs. Cervical MRI was carried out randomly and showed abnormal findings in 21 of 25 patients, in whom 18 duodenal tissues were examined by biopsies. Copyright (C) 2002 S. Karger AG, Basel.

Mice immunized with fibroblasts expressing an MHC class II molecule and human thyrotropin receptor (TSHR), but not either alone, develop major features characteristic of Graves' disease (GD), such as thyroid-stimulating autoantibodies directed against TSHR, increased serum thyroid hormone levels, and enlarged thyroid glands. The results indicate the need for the simultaneous expression of a class II molecule and the TSHR on the surface of the fibroblasts to develop stimulating anti-TSHR antibodies and full-blown GD in our model. A T cell line established from a mouse with hyperthyroidism proliferates in response to fibroblasts expressing a class II molecule and TSHR, but not to the fibroblasts expressing only TSHR, indicating that the class II molecules on the fibroblasts present TSHR-derived peptide(s) to T cells. These results strongly suggest that the acquisition of antigen-presenting ability by thyrocytes can lead to the induction or progression of GD. We identified a T cell epitope of TSHR by the proliferative response of spleen cells from mice immunized with fibroblasts expressing a class II molecule and TSHR to 80 overlapping peptides spanning the extracellular domain of human TSHR. The identification of a major T cell epitope provides an important clue to a novel therapy of GD.

Sjogren's syndrome (SS) is thought to be uncommon in children. We studied the clinical manifestations and laboratory findings of 12 pediatric patients with SS, all of the children did not have sicca symptoms but have lymphocytic infiltration of salivary glands, abnormal sialograms or abnormal results of scintigraphy compatible with typical SS. Seven cases had primary SS and five were secondary SS and had other autoimmune disorders (three cases with systemic lupus erythematosus, one case with dermatomyositis, and the other with mixed connective tissue disease). All patients were female. The mean age at onset of symptoms, including other autoimmune manifestations, was 12.2 years (range 9-15 years). The initial symptoms were some systemic manifestations (fever, exanthema, arthralgia, etc.) and various autoimmune phenomena (butterfly rash, Raynaud's phenomenon, proteinuria, weakness of muscles, etc.). On the other hand, no patients complained of sicca symptoms. Laboratory studies in our patients revealed elevated levels of IgG (92%), antinuclear antibody (92%), rheumatoid factor (58%), anti-SS-A antibody (75%). These findings were similar to those found in adult patients with sicca symptoms previously reported in literature. From these studies, we suggest that lip biopsy, sialography and/or salivary gland's scintigraphy should be carried out in patients who had abnormal laboratory findings as mentioned above, irrespective of absence of sicca symptoms, in order to diagnose SS at early period.