Association of the MMP9 gene with childhood cedar pollen sensitization and pollinosis.

Matrix metalloproteinase 9 (MMP9) gene has been shown to be involved in the pathogenesis of allergic rhinitis (AR) and asthma. Previous studies suggested that single-nucleotide polymorphisms (SNPs) of the MMP9 gene conferred a risk for childhood asthma. However, whether the SNPs confer a risk for AR has not been previously investigated. The objective of this study was to investigate whether SNPs of the MMP9 gene are associated with risk of seasonal AR (pollinosis), perennial AR and allergen sensitization. A total of 670 school children were recruited in Japan and genotyped for functional polymorphism in the promoter (-1590C/T: rs3918242) and three amino-acid substitutions (R297Q: rs17576; P574R: rs2250889; R668Q: rs17577). Serum levels of total and specific IgE were determined. Disease status and other clinical characteristics of the subjects were investigated using a questionnaire. Associations between the MMP9 SNPs and both AR and serum IgE levels were evaluated. -1590C/T showed significant association with cedar pollinosis (corrected P (Pcor)=0.039). R668Q was in strong linkage disequilibrium (LD) with -1590C/T and showed significant association with cedar pollinosis (Pcor=0.023) and serum cedar pollen-specific IgE level (Pcor=0.022). A haplotype associated with -1590T and 668Q showed a significant association with cedar pollinosis, orchard grass pollinosis and cedar pollen-specific IgE (Pcor=0.0012, Pcor=0.0059 and Pcor=0.0041, respectively). R297Q and P574R were in weak LD with the rest of the SNPs and did not show significant association with disease. Compared with wild-type MMP9 protein (279R-574P-668R), a variant enzyme (279R-574P-668Q) that showed association with pollinosis had lower activity. However, lower enzyme activity was not associated with disease risk because another variant (279Q-574R-668R) showed lower enzyme activity but was not associated with pollinosis. The -1590T allele and its corresponding haplotype was associated with higher promoter activity and with pollen-specific IgE levels and pollinosis, suggesting that -1590C/T may have more impact on sensitization and disease development than R668Q. Our results suggest that the MMP9 gene confers susceptibility to cedar pollinosis in Japanese children. The MMP9 gene may be associated with pollinosis through sensitization processes.

Aldose reductase deficiency in mice protects from ragweed pollen extract (RWE)-induced allergic asthma.

BACKGROUND: Childhood hospitalization related to asthma remains at historically high levels, and its incidence is on the rise world-wide. Previously, we have demonstrated that aldose reductase (AR), a regulatory enzyme of polyol pathway, is a major mediator of allergen-induced asthma pathogenesis in mouse models. Here, using AR null (AR-/-) mice we have investigated the effect of AR deficiency on the pathogenesis of ragweed pollen extract (RWE)-induced allergic asthma in mice and also examined the efficacy of enteral administration of highly specific AR inhibitor, fidarestat. METHODS: The wild type (WT) and AR-/- mice were sensitized and challenged with RWE to induce allergic asthma. AR inhibitor, fidarestat was administered orally. Airway hyper-responsiveness was measured in unrestrained animals using whole body plethysmography. Mucin levels and Th2 cytokine in broncho-alveolar lavage (BAL) were determined using mouse anti-Muc5A/C ELISA kit and multiplex cytokine array, respectively. Eosinophils infiltration and goblet cells were assessed by H&E and periodic acid Schiff (PAS)-staining of formalin-fixed, paraffin-embedded lung sections. T regulatory cells were assessed in spleen derived CD4+CD25+ T cells population. RESULTS: Deficiency of AR in mice led to significantly decreased PENH, a marker of airway hyper-responsiveness, metaplasia of airway epithelial cells and mucus hyper-secretion following RWE-challenge. This was accompanied by a dramatic decrease in infiltration of eosinophils into sub-epithelium of lung as well as in BAL and release of Th2 cytokines in response to RWE-challenge of AR-/- mice. Further, enteral administration of fidarestat significantly prevented eosinophils infiltration, airway hyper-responsiveness and also markedly increased population of T regulatory (CD4+CD25+FoxP3+) cells as compared to RWE-sensitized and challenged mice not treated with fidarestat. CONCLUSION: Our results using AR-/- mice strongly suggest the role of AR in allergic asthma pathogenesis and effectiveness of oral administration of AR inhibitor in RWE-induced asthma in mice supports the use of AR inhibitors in the treatment of allergic asthma.

Spring brings breezes, wheezes, and pollen oxidases.

While the release of pollen into the air is essential for the reproduction of plants, the accidental yet inevitable uptake of pollen into human airways can cause symptoms of seasonal allergies and asthma. The symptomatic response to pollen is caused by granulocytes that produce inflammation, which is due in part to oxidative stress through the action of NADPH oxidases. The recruitment of these inflammatory granulocytes was previously thought to depend entirely on the activation of an adaptive immune response. In this issue of the JCI, Boldogh et al. demonstrate that pollens contain endogenous NADPH oxidase activity, which functions to generate local "danger signals" in nearby airway epithelium. These signals in turn trigger the early recruitment of granulocytes, even in the absence of the adaptive immune response. These findings suggest that inhibition of the pollen oxidase may provide a way to antagonize allergic inflammation at a very early step.

The urokinase system in patients with intermittent and persistent allergic rhinitis.

Local and systemic changes in hemostasis are associated with allergic diseases. Apart from their well documented role in regulation of plasminogen activation, components of the urokinase system may be involved in modulation of cellular activities during immune inflammatory responses. So far, little has been known about the function of the system in allergic inflammation. In the present study, we assessed circulating levels of the urokinase system components such as urokinase-type plasminogen activator (uPA), soluble form of uPA receptor (CD87), and the inhibitor--plasminogen activator inhibitor type 1. The study comprised of patients suffering from allergic rhinitis, however, without any asthmatic symptoms. Plasma levels of uPA and soluble form of uPA receptor antigens, and plasminogen activator inhibitor type 1 activity were measured in 17 patients with grass pollens-induced intermittent allergic rhinitis and 15 patients with persistent allergic rhinitis due to house dust mite allergy, as well as in 20 sex-matched and age-matched healthy nonatopic participants. We did not observe any statistically significant differences in the levels of the urokinase system components between patients with intermittent allergic rhinitis and persistent allergic rhinitis, and the controls. The circulating levels of uPA, its soluble receptor, and its inhibitor did not differ between allergic rhinitis patients and healthy participants, therefore it seems that the systemic release and activity of the urokinase system molecules may be not significantly changed in the course of nasal allergic inflammation induced by periodic or continuous exposure to a natural allergen.

Monitoring antioxidant enzymes in red cells during allergen immunotherapy.

The aim of this report was to answer the question how specific immunotherapy influences the antioxidant enzyme system in patients with respiratory allergy and in longer perspective to find markers suitable to assess the efficacy of treatment. In open prospective randomised study 28 patients (18 females and 10 males, age 14-48 years) with seasonal respiratory allergy were treated with allergen immunotherapy. Subjects received subcutaneous therapy with allergens absorbed on calcium phoshate or aluminium hydroxide and were analyzed by the established protocol at the beginning, after three and 12 month of the treatment. In all treatment group red cell superoxide dismutase and glutathione peroxidase activities were in the normal range in allergic patients both before and during the treatment. Catalase activity in the allergic patients was lower as compared with controls and a significant increase of the enzyme activity occurred during and at the end of the treatment. In patients treated with calcium phosphate adsorbed allergen there was a continous increase of catalase activity from beginning up to the end of observation. In the case of the aluminium hydroxide treatment there was an increase from the baseline values up in the third month of the treatment and a decrease on the 12th month. In summary, the present results open the question that allergen immunotherapy may cause imbalance of oxidants and antioxidants. To support our findings larger controlled field studies are needed.

Metachromatic, IgE-bearing and tryptase-containing cells on the nasal mucosal surface in provoked allergic rhinitis.

We studied the relationship between mast cells or basophils and symptoms in provoked allergic rhinitis. Nasal brush and lavage samples were obtained before nasal allergen challenge and every 2 h for 12 h after the challenge in 10 allergics and 3 controls. The cells were identified by their metachromatic staining properties (brush and lavage samples) or with immunohistochemical methods using specific antibodies to IgE and tryptase, a selective mast-cell marker (brush samples). Histamine was determined in the brush samples using liquid chromatography. After an initial decrease, the numbers of metachromatic, IgE-bearing and tryptase-containing cells, as well as the histamine content of the cells in the brush samples, increased during the subsequent observation hours. The prechallenge cell and histamine values correlated with the symptom score 15 min after the challenge. The prechallenge lavage samples lacked metachromatic cells, but these cells were found in increasing numbers after the provocation. Three of the patients differed from the remaining seven in that their prechallenge brush samples contained many positively stained cells. All patients showed a positive cellular response to the allergen challenge, but these three individuals showed the most vivid response. The morphology of the metachromatic cells in the prechallenge brush samples agreed with that of mast cells, but the morphology of metachromatic cells which outnumbered tryptase-containing cells in samples at 8 to 12 h rather agreed with their being basophils.

Acetylcholinesterase levels in nasal turbinate congestion.

In a previous communication, one of the authors discussed prolonged congestion of the turbinates following nasal surgery. The clinical factors responsible were allergy or the traumatic effects of nasal packing on the turbinates. A study of turbinate function was done to find the factor responsible for this congestion. Biopsies of an inferior turbinate were obtained preoperatively and two weeks after surgery. The specimens were examined for the level of acetylcholinesterase by histochemical assay and were also studied by examining sections histologically. In the majority of cases, the level of acetylcholinesterase fell with the appearance of congestion and rose when the turbinates returned to normal. These results suggest a connection between turbinate congestion and levels of tissue acetylcholinesterase in the presence of inflammation or allergy.

Evaluation of a bedtime dose of a combination antihistamine/analgesic/decongestant product on antigen challenge the next morning.

The effects of CAAD (diphenhydramine hydrochloride, 50 mg; pseudoephedrine hydrochloride, 60 mg; acetaminophen, 1 g in 10% ethanol) were evaluated in a double-blind, three-way, placebo-controlled, cross-over study on 18 volunteers with allergic rhinitis. The number of sneezes following nasal challenge with antigen was significantly reduced after a bed-time dose of CAAD (P less than .005) and a single dose of diphenhydramine (P less than .001) given 2 hours before the challenge. The levels of N-alpha-tosyl-L-arginine methyl ester (TAME) activity decreased after diphenhydramine treatment, while histamine levels following challenge were not different. The drowsiness reported after CAAD was equal to placebo, but significantly less than diphenhydramine (P less than .002 for both). The active treatments reduced the actions of histamine without suppressing its release from mast cells. The effect of CAAD persists 10 hours after administration without inducing drowsiness.

[The pathogenesis of nasal mucosa polyps, studied with histochemical detection of phenol oxidase (E. C. 1.14.18.1)]. / Zur Pathogenese von Nasenschleimhautpolypen, untersucht mit dem histochemischen Nachweis für Phenoloxidase (EC 1.14.18.1).

There were investigated polyps of the nasal mucous membrane with the histological diagnosis: allergic-hyperergic, lympho-plasma cellular, and angioma-like. Using the phenol oxidase reaction, it was possible to integrate the allergic-hyperergic and the lympho-plasma cellular polyps into the pathologic course of cell mediated immunological defence reactions in which the eosinophilic granulocytes have an essential role. The eosinophils are typical for the immunological character of the defense reaction.

The effect of antihistamines on the immediate allergic response: a comparative review.

Antihistamines are believed to reduce the sneezing and rhinorrhea associated with allergic rhinitis, primarily by competitive antagonism of histamine for H1 cellular receptors, but additional mechanisms of action may contribute to their clinical efficacy. To improve our understanding of H1 antihistamine action, we studied the effects of pretreatment with terfenadine, cetirizine, ketotifen, azatadine, diphenhydramine, and azelastine on increases in vascular permeability, mast cell activation, and sneezing induced by nasal challenge with antigen. All studied antihistamines reduced sneezing, indicating that they all effectively antagonize histamine after its release. In addition, terfenadine and topically administered azatadine blocked the release of histamine. Studies with cetirizine and azelastine revealed that these antihistamines significantly reduced sulfidopeptide leukotriene levels. Terfenadine and azelastine also reduced kinin production. These results confirm that antihistamines are effective in reducing sneezing and, in some cases, vascular permeability. The findings of these studies also illustrate that the various antihistamines have multiple and different mechanisms of action that may have implications for their clinical uses.

Changes of serum histaminase activity in pollinosis.

In order to study changes in serum histaminolytic enzyme activity in allergic individuals following allergen challenge, serum histaminase activity was measured using the 0-dianisidine peroxidase method in patients with Japanese cedar pollinosis. Significantly higher serum histaminase activity was detected in pollen season than out of season and a significant relationship was recognized between serum histaminase activity and the severity of nasal symptoms.

[Levels of interleukin-4, interleukin-5, tryptase and eosinophil cationic protein of nasal lavage fluid in pollen allergic rhinitis]. / Interleukin-4, interleukin-5, triptáz és eosinophil kationos proteinszintek meghatározása az orrmosó folyadékban rhinitis allergicában.

IL-4, IL-5, tryptase and eosinophil cationic protein levels were measured in nasal lavage fluid from 15 pollen allergic rhinitis beyond pollen season. Allergy was proved by prick test. There were 15 non allergic children in the control group. Specific nasal allergen provocation was performed on the rhinitic group. Nasal lavage were done before, 1 and 12 hours after the provocation. Before the nasal provocation the ECP and IL-4 levels were significantly higher in the allergic group compared to the non allergic group. The levels of tryptase, ECP and IL-4 rose significantly after the provocation. The results reflect to the possibility of an activated immune status in allergic rhinitis even without the presence of the triggering pollens. After the specific provocation elevated tryptase levels were measured, referring to the activity of the early phase of the I. type hypersensitivity reaction, while the ECP and IL-5 elevation to its late phase. According to our examinations it can be said, that tryptase, ECP and IL-5 might be used to detect the activation of the early and late phases of the IgE mediated hypersensitive reaction.

[Activity of glucose-6-phosphate dehydrogenase in erythrocytes in patients with atopic asthma and allergic rhinitis]. / Aktywnosc dehydrogenazy glukozo-6-fosforanowej krwinek czerwonych u chorych na atopowa astme oskrzelowa i katar sienny.

A significant decrease of erythrocyte G-6-P dehydrogenase activity in patients with atopic asthma and allergic rhinitis was observed. In order to define the cause of this phenomenon the enzyme was isolated from the cells. The kinetic and molecular properties of the erythrocyte G-6-P dehydrogenase were studied. A change in the Michaelis constant for G-6-P dehydrogenase was seen in erythrocytes of both patient groups. The thermoresistance at 45 C was reduced significantly. These modified properties can imply a change in the dehydrogenase molecule, this is seen more significantly in the group with allergic rhinitis.

[Myeloperoxidase (MPO) as a marker of neutrophil influx into nasal mucosa after recombinant IL-8 challenge]. / Mieloperoksydaza (MPO) jako marker naplywu neutrofilów do blony sluzowej nosa po prowokacji rekombinowana Il-8.

Myeloperoxidase (MPO) has been proposed to mirror the degree of neutrophil activation, however its role as a marker of the participation of neutrophils in allergic inflammation is still unclear as the literature is controversial. The aim of this study was to evaluate the MPO levels and neutrophil influx in nasal lavage after recombinant Il-8 challenge. Eight patients suffering from seasonal allergic rhinitis, hypersensitive to grass pollens, with average age 30.1 +/- 2.67 were challenged both with Il-8 and diluent for Il-8 on a subsequent day, in two phases: before the pollen season (unprimed mucosa) and during the season (primed mucosa). Nasal lavage with saline were collected before, during Il-8 or placebo challenge and 30 minutes, 2 hours and 3 hours after the challenge. The number of neutrophils and MPO levels in the nasal fluid were determined. After the challenge with Il-8 of primed mucosa the number of neutrophils increased from 28250 cells/ml at the baseline to 28778, 251020 and 333660 at 0, 5, 2 and 3 hours respectively. There was a statistically significant difference between cells number after diluent and Il-8 challenge (p < 0.05, Wilcoxon rank-sum test). The number of neutrophils in the nasal lavage of primed mucosa after Il-8 challenge was significantly higher (p < 0.005) at all time points in comparison with the diluent and Il-8 challenges in the unprimed mucosa. There was no difference (p < 0.05) in MPO levels in the nasal lavage between Il-8 (mean 17.43 ng/ml +/- 10.98) and diluent challenge (20.98 ng/ml +/- 11.89) of unprimed mucosa. In the primed mucosa fluid we observed a peak of MPO level at 2 hours time point, however that was not significant as compared to diluent challenge (p = 0.465). We did not find the relationship between MPO levels and the neutrophils number in the lavage (rank Spearman factor, rs = 0.258, p = 0.42). Due to the lack of statistically confirmed relationship between MPO level and the number of neutrophils, MPO seems to be of little value as a marker of neutrophil influx into nasal mucosa.

[Alpha 1-protease inhibitor: characteristics of its biochemical and biological properties and its level in various diseases (review of the literature and personal observations)]. / Alpha 1-ingibitor proteaz: kharakteristika biokhimicheskikh i biologicheskikh svoistv i opredelenie urovnia pri razlichnykh zabolevaniiakh (obzor literatury, sobstvennye nabliudeniia).

The authors review biological and biochemical properties and the clinical importance of the serum proteases alpha 1-inhibitor with broad-range antiproteolytic activity. Congenital deficiency of this protein is a frequent enough condition linked with predisposition to some diseases of the lungs and liver. Early determination of the deficiency of alpha 1-Pi is fairly urgent, since it permits the early administration of the preventive measures and substitution therapy. The immunochemical technique makes it possible to determine all the varieties of alpha 1-Pi, as they are antigenically similar. Sera from 267 patients with different diseases were examined. The content of alpha 1-Pi was found to be elevated in Waldenström's macroglobulinemia, chronic active hepatitis and liver cirrhosis and to be lowered in bronchial asthma. In multiple myeloma and pollinoses, no alterations in the alpha 1-Pi content were recorded.

Prevention of allergic rhinitis by aldose reductase inhibition in a murine model.

BACKGROUND: Allergic rhinitis, one of the most common atopic diseases, is known to be elicited by Th2 cytokine-mediated inflammatory response. We have shown earlier that a polyol pathway enzyme aldose reductase (AR) regulates airway inflammation; however its role in allergic rhinitis is not known. We have investigated the role of AR in mediating pathological symptoms associated with allergic rhinitis in mice. METHODS: The wild-type (WT) mice treated without or with AR inhibitor and AR knock out (AR(-/-)) mice were sensitized by two intraperitoneal injections of ragweed pollen extract (RWE) with adjuvant alum on days 0 and 4 followed by challenge on day 11 and/or 18 and 25. The allergic rhinitis symptoms were assessed by monitoring the nasal scratch, mast cell degranulation and release of tryptase in nasal lavage, infiltration of inflammatory cells, production of inflammatory cytokines and nasal epithelium remodeling. RESULTS: Sensitization and challenge of mice with RWE produced robust and reproducible pathological symptoms of allergic rhinitis as compared to control mice. AR inhibitor, fidarestat administered mice showed markedly reduced early phase response to allergen exposure such as nasal scratches, mast cells degranulation and release of tryptase in the nasal passage as well as late phase response such as inflammatory cell infiltration and release of Th2 type cytokines and nasal epithelial remodeling. Further, prevention of these events in AR(-/-)) mice suggests the role of AR in the mediation of allergic rhinitis. CONCLUSION: These results indicate an important role of AR in the mediation of RWE-induced allergic rhinitis in mice and prevention by AR inhibitor, fidarestat offers a novel therapeutic approach to ameliorate allergic rhinitis.