Hiroko Watanabe

Circadian Variation in Nasal Reactivity in Children with Allergic Rhinitis: Correlation with the Activity of Eosinophils and Basophilic Cells

Background: In allergic rhinitis, the major symptoms of runny nose, sneezing, and stuffy nose tend to become worse upon waking up in the morning, and yet the mechanisms underlying this phenomenon are poorly understood. We investigated whether the worsening of allergic rhinitis in the morning is associated with changes in the activity of inflammatory cells. Methods: Nasal reactivity to methacholine was assessed twice in 8 children with allergic rhinitis and 8 healthy control subjects at 6.00 a.m. and 3.00 p.m. The amounts of eosinophil cationic protein (ECP), histamine and tryptase in induced nasal secretions and peripheral blood were also measured. Results: Nasal reactivity to methacholine was higher at 6.00 a.m. not only in patients but also in healthy controls. Serum ECP and plasma histamine levels showed no circadian patterns. On the other hand, significantly higher levels of inflammatory activation products were found in nasal secretions at 6.00 a.m., thus showing a direct association with nasal reactivity. Conclusion: These results suggest that the circadian variation in nasal reactivity is associated with changes in the activity of eosinophils and basophilic cells in the nasal mucosa.

Heat treatment of egg white controls allergic symptoms and induces oral tolerance to ovalbumin in a murine model of food allergy

SCOPE Heated foods often present low allergenicity, and have recently been used in specific oral immunotherapy for food allergies. However, the influence of heating on tolerogenicity of food allergens is not well elucidated. Here, we investigated biochemical, allergenic, and tolerogenic properties of heated egg white (EW) using a murine model of food allergy. METHODS AND RESULTS Raw EWs were treated at 80°C for 15 min (80EW, mild heating condition), 100°C for 5 min (100EW, cooking condition), or 121°C for 40 min (121EW, retort pouch condition), and freeze-dried. A transgenic OVA23-3 mice model expressing T-cell receptor specific for ovalbumin (OVA, a major EW allergen) induced Th2 cells and IgE production, and presented intestinal inflammation when fed untreated EW diet. 80EW-fed mice presented only moderate inflammation but high Th2 responses. 100EW-fed mice did not present inflammation but induced tolerance as seen in reduced T-cell responses and IgE levels. 100EW demonstrated higher digestive stability and slower absorption in intestine, compared with untreated EW and 80EW. 121EW was strongly aggregated, was not absorbed well, and developed Th1 responses without tolerance induction. CONCLUSION OVA in EW treated only under a particular heat condition (e.g. 100°C for 5 min) lost allergenicity, but possessed tolerogenicity.

Functional variants in the thromboxane A2 receptor gene are associated with lung function in childhood‐onset asthma

The thromboxane A2 receptor (TBXA2R) gene is associated with asthma, but no functional genetic variations are known to associate with the disease or its related phenotypes.

Induction of the Matrix Metalloproteinase 13 Gene in Bronchial Epithelial Cells by Interferon and Identification of its Novel Functional Polymorphism.

Matrix metalloproteinases (MMPs) are a class of extra-cellular and membrane-bound proteases involved in a wide array of physiological and pathological processes including tissue remodeling, inflammation, and cytokine secretion and activation. MMP-13 has been shown to be involved in lung diseases such as acute lung injury, viral infections, and chronic obstructive pulmonary disease; however, the molecular pathogenesis of MMP-13 in these conditions is not well understood. In this study, we investigated the mechanisms and roles of MMP-13 secretion in human small airway epithelial cells (SAECs) and functional polymorphisms of the MMP13 gene. Polyinosinic–polycytidylic acid (poly(I:C)) and interferon β (IFN-β) stimulated the secretion of MMP-13 from SAECs by more than several hundred-fold. Stimulation of the secretion by poly(I:C) was abolished by SB304680 (p38 inhibitor), LY294002 (PI3K inhibitor), Janus kinase (JAK) inhibitor I, RNA-activated protein kinase (PKR) inhibitor, and Bay 11-7082 (NF-κB inhibitor), while stimulation by IFN-β was inhibited by all except Bay 11-7082. These data suggested that the secretion of MMP-13 was mediated through IFN receptor pathways independently of nuclear factor kappa B (NF-κB) and that poly(I:C) stimulated IFN secretion in an NF-κB-dependent manner from SAECs, leading to IFN-stimulated MMP-13 secretion. Chemical MMP-13 inhibitors and MMP-13 small interfering RNA (siRNA) inhibited IFN-stimulated secretion of interferon gamma-inducible protein 10 (IP-10) and regulated on activation, normal T-cell expressed and secreted (RANTES), suggesting that MMP-13 is involved in the secretion of these virus-induced proinflammatory chemokines. We identified a novel functional polymorphism in the promoter region of the MMP13 gene. The MMP13 gene may play important roles in defense mechanisms of airway epithelial cells.

Association Study of Matrix Metalloproteinase-12 Gene Polymorphisms and Asthma in a Japanese Population

Background: Matrix metalloproteinase 12 gene (MMP12) has been shown to be associated with asthma in a Caucasian population. In this study, we investigate whether single-nucleotide polymorphisms (SNPs) of MMP12 are associated with a risk for asthma in a Japanese population. Methods: We tested for an association between SNPs in MMP12 and asthma, including its severity, in a Japanese population (630 pediatric and 417 adult patients with atopic asthma and 336 children and 632 adults as controls). The rs652438 A and G variants (N357S) were generated by site-directed mutagenesis and an assay with artificial peptide substrates was used to compare two types of MMP12 activity. The effect of MMP12 inhibition with MMP12-specific small interfering RNA (siRNA) on chemokine secretion from airway epithelial cells was also tested in vitro. Results: N357S showed a p value <0.05 for childhood and combined (adult plus childhood) asthma in the dominant model [odds ratio (OR) 1.60, 95% confidence interval (CI) 1.00–2.56, p = 0.047; OR 1.40, 95% CI 1.04–1.89, p = 0.028, respectively]. This risk variant is associated with asthma severity in adult patients. In the functional assay, the minor-allele enzyme showed significantly lower activity than the major-allele enzyme. MMP12-specific siRNA suppressed IP-10 secretion from airway epithelial cells upon stimulation with IFN-β. Conclusions: Our results suggest that MMP12 confers susceptibility to asthma and is associated with asthma severity in a Japanese population. MMP12 may be associated with asthma through inappropriate attraction of leukocytes to the inflamed tissue.

Effects of Chemicals on the Immune Response

We have tested the effects of various environmental chemicals on proliferation of murine antigen-primed lymphocytes and their secretion of cytokines. Lymph node (LN) cells from mice immunized with bovine αsl -casein were incubated with this antigen in the presence of β-benzene hexachloride (β-BHC), p,p′-dichlorodiphenyldichloroethylene (p,p,p′-DDE), p-nitrophenyl 2, 4, 6-trichlorophenyl ether (CNP), D-α-aminobenzylpenicillin (Amp), and o-acetylcitric acid tributyl ester (ATBC), and tested for proliferation and secretion of interleukin (IL-) 2, IL-4, IL-10 and interferon γ. The results of the proliferation assays indicated that more than 20 ppm of CNP and DDE strongly inhibited the proliferation of LN T cells. All of the chemicals tested affected cytokine secretion by LN cells in response to antigen. The effects of these chemicals on cytokine secretion varied depending upon the cytokine examined and were independent of the inhibitory effect on cell proliferation. These results suggest that costimulation of T lymphocytes with certain environmental chemicals can modulate the antigen-specific cytokine responses of these cells, even at concentrations which are considered to have no toxicity and no inhibitory effect on the proliferative response of T cells.

Interleukin-10 and interleukin-5 balance in patients with active asthma, those in remission, and healthy controls

Background The immunological mechanisms of asthma remission remain unclear although several reports have suggested that balance between T helper (Th) 2 cytokines and regulatory cytokines is related. Objective To study the balance between interleukin (IL) 10 and IL-5 in asthma clinical remission. Methods We measured the numbers of IL-5 and IL-10 producing cells in peripheral blood mononuclear cells stimulated with mite antigen obtained from patients with active asthma (group A, n = 18), patients in clinical remission (group R, n = 15) and nonatopic healthy controls (group H, n = 14). Results The numbers of IL-5 producing cells in groups A and R were significantly higher than in group H. The number of IL-5 producing cells was lower in group R than in group A, although the difference was not statistically significant. The number of IL-10 producing cells was higher in group R than in group A, although again the difference was not statistically significant. There was a significant difference in the number of IL-10 producing cells between groups A and H but not between groups R and H. The ratio of the number of IL-10 to IL-5 producing cells was highest in group H followed by groups R and A, and the differences were statistically significant for each pair of groups. Conclusion Our study suggests that the IL-10/IL-5 balance is related to clinical asthma. The balance differs between patients in clinical remission and healthy controls, suggesting that allergic inflammation may continue even after clinical asthma remission.

Environmental tobacco exposure is associated with vaccine modified measles in junior high school students

Vaccine modified measles (VMM) affects individuals with attenuated vaccine induced immunity. An outbreak of measles occurred in a junior high school, starting from an unvaccinated eighth‐grade student who developed natural measles and affected a majority of students who were immunized with a low potent strain of measles vaccine (TD97). To determine whether environmental tobacco smoke (ETS) exposure was associated with the development of VMM in this population, a questionnaire was used asking whether students had VMM symptoms during the outbreak and the smoking status of family members. VMM was defined in the study population as occurrence of fever and/or erythema, along with documented history of measles vaccination. A total of 513 students (85.9%) responded. Overall, the presence of in‐house smokers did not differ between VMM students (49.3%) and non‐VMM students (50.2%). However, in the ninth grade, presence of an in‐house smoker was significantly higher in the family of VMM students (54.0%) than in non‐VMM students (36.6%) (P = 0.044). Urinary cotinine levels were also measured in selected students (n = 37). Among families with at least one smoker, urinary cotinine levels were significantly higher in VMM students than in non‐VMM students (P = 0.032). Furthermore, a multivariable logistic regression analysis showed that a high urinary cotinine level (>10 ng/mg creatinine; 13.5 percentile) was associated with the development of VMM. Our findings suggest that a high level of ETS exposure may be associated with an increased risk of VMM in a population with attenuated vaccine induced immunity against measles. J. Med. Virol. 87:1853–1859, 2015.