bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of eosinophils and detection of eotaxin in skin chamber fluid after challenge with relevant allergen in patients with mild asthma
Authors: Fernvik  , Grönneberg  , Lundahl  , Raud  , Zetterström  , Van Hage‐Hamsten  , Halldén  
Published: March 2003

BackgroundA selective recruitment of eosinophils to sites of allergic inflammation is suggested to be controlled by regulation of cytokines, chemokines and adhesion molecules.ObjectiveThe aim of this study was to examine whether allergen challenge in skin chambers, applied on patients with allergic rhinitis and mild asthma, results in a selective influx of activated eosinophils and detectable levels of cytokines/chemokines related to eosinophil recruitment, such as interleukin (IL)‐5 and eotaxin.MethodsA skin blister was induced on the volar aspect of each forearm; one contained PBS‐heparin buffer (control) and the other was challenged with relevant allergen. Peripheral blood was drawn before the allergen was applied to the skin chamber, and the expression of CD9, CD11b and EG2‐epitope on intracellular eosinophil cationic protein (ECP) was analysed in eosinophils. Chamber fluid was collected 8 h after allergen application and analysed for differential cell counts, expression of eosinophil activity markers, the presence of ECP, eotaxin, and IL‐5.ResultsThe number of recruited leucocytes was equal in the allergen‐challenged chambers and in controls. However, the number of eosinophils was significantly increased in the allergen‐challenged chambers, and elevated levels of released ECP were measured. Moreover, the eosinophils recruited were activated, as shown by increased expression of EG2 and CD11b, and decreased expression of CD9, in comparison with blood eosinophils. In the skin chamber fluids, higher levels of eotaxin were detected in the allergen‐challenged chambers than in controls, but there were no detectable levels of IL‐5.ConclusionWe have demonstrated a selective recruitment of eosinophils, and higher levels of released ECP and eotaxin, in skin chambers stimulated with allergen, as compared with control chambers. Allergen challenge in skin chambers is a useful tool for studies of eosinophil recruitment, their state of activation, and their involvement in the allergic inflammatory response.
Nasal response to a single antigen challenge in patients with allergic rhinitis — inflammatory cell recruitment persists up to 48 hours
Authors: MIADONNA  , MILAZZO  , GIBELLI  , SALMASO  , LORINI  , TEDESCHI  
Published: March 2003

BackgroundAllergen challenge in some patients with respiratory allergy is followed by an early and a late reaction.ObjectiveTo evaluate the duration of mediator release and inflammatory cell recruitment during the late antigen‐induced nasal response.MethodsEight patients with seasonal allergic rhinitis due to grass pollen underwent local challenge with the relevant allergen, a non‐relevant allergen (Parietaria judaica), and nebulized saline solution. Nasal lavages were performed at baseline and 6, 24, 48, 72 h after challenge. Eosinophil cationic protein (ECP), leukotriene C4 (LTC4), leukotriene B4 (LTB4) myeloperoxidase (MPO) and prostaglandin D2 (PGD2) levels were radioimmunoassayed and histamine concentration was measured by an automated fluorometric method.ResultsNasal challenge with the relevant antigen induced a response 6 h after stimulation, which subsided within 24 h. Eosinophilia, observed in the nasal lavages collected from 6 to 24 h after this challenge, was accompanied by ECP release. Neutrophilia were found in the nasal lavages collected from 6 to 24 h after challenge. The increase in neutrophil number correlated with MPO levels and LTB4 concentrations, but not with the intensity of nasal obstruction. Antigen challenge also induced significant recruitment of mononuclear cells 48 h after provocation. The challenge significantly raised histamine, but not PGD2, levels in the nasal lavages collected 6 h after provocation. A trend towards an increase in LTC4 levels in the nasal lavages collected 6 h after specific antigen challenge was also found. Nasal challenge with a non‐relevant allergen or with saline solution did not cause either inflammatory cell recruitment or mediator release.ConclusionNasal challenge with the relevant antigen can induce a late response characterized by local accumulation of eosinophils, neutrophils and mononuclear cells persisting for 48 h and accompanied by release of ECP, MPO, LTB4 and histamine. These results indicate that a single antigen challenge in patients with allergic rhinitis causes prolonged inflammatory alterations which may contribute to the development of airway hyperreactivity.
Identification and characterization of the major allergen of the Humulus japonicus pollen
Authors: Park  , Ko  , Kim  , Jeoung  , Hong  
Published: March 2003

BackgroundPollen of Humulus japonicus has been known as one of the important causes of pollinosis in Korea and China. To date, the major allergen of H. japonicus has not been determined.ObjectiveTo identify the major allergen of H. japonicus pollen and characterize its biochemical properties.MethodsWith the sera of 29 patients reactive to H. japonicus, the major allergen of H. japonicus was determined from the results of IgE immunoblotting and ELISA inhibition. The biochemical properties of the major allergen of H. japonicus were evaluated by lectin blotting assay and 2‐dimensional PAGE blot. N‐terminal amino acid sequences were determined by the Edman degradation method. The suggested major allergen was purified by DEAE anion exchange and gel filtration chromatography.ResultsTwenty‐nine sera contained IgE bound to the 10, 16, 20, 29 and 42 kDa proteins of H. japonicus in immunoblot analysis. A protein of 10 kDa was the most prevalent allergen in the sera of H. japonicus‐reactive patients (72%). The ELISA optical density of H. japonicus‐specific IgE was not inhibited by pollen extracts of birch, oak, rye grass and mugwort. The 10‐kDa allergen was neither stained with PAS nor bound with ConA and five other lectins. The isoelectric point of the 10‐kDa allergen was approximately pH 5.1. We sequenced the N‐terminal amino acids of the 10‐kDa allergen, which was not homologous with any previously characterized allergen. The 10‐kDa allergen could be purified with DEAE anion exchange and gel filtration chromatography. Maximum inhibitions of H. japonicus‐specific IgE ELISA by whole extract of H. japonicus and purified 10‐kDa allergen were more than 97 and 88%, respectively, while the 50% inhibitory concentration of the whole extract of H. japonicus and purified 10 kDa were 38 and 20 ng/mL, respectively.ConclusionThe 10‐kDa peptide could be a major allergen of H. japonicus. Its isoelectric point was 5.1 and it did not bind with lectins. The N‐terminal amino acid sequence of the 10‐kDa major allergen was also determined.
Sensitization to grass, ragweed, mugwort pollen allergens in Japanese monkeys (Macaca fuscata): preliminary report
Authors: Sakaguchi M. , Kobayashi C. , Inouye S. , Konaka A. , Yamada T. , Nigi H. 
Published: March 2003

BackgroundThe natural occurrence of Japanese cedar (CJ, Cryptomeria japonica) pollinosis has been reported in Japanese monkeys (Macaca fuscata). Furthermore, most of these monkeys with CJ pollinosis have immunoglobulin (Ig) E sensitization to Japanese cypress (Chamaecyparis obtusa) pollen. However, specific IgE to other pollens has not yet been reported.ObjectivesThe present study was designed to investigate IgE sensitization of Japanese monkeys to grass, ragweed, and mugwort pollen.MethodsSerum samples from 47 monkeys as a general population in one troop were collected at random. We measured specific IgE to grass, ragweed and mugwort pollen. Next, 10 monkeys with CJ pollinosis from the same troop were also examined for their IgE sensitization to grass, ragweed, and mugwort pollen.ResultsOf 47 monkeys, 13 (28%) had specific IgE to CJ pollen, 15 (32%) to grass pollen, five (11%) to ragweed pollen, and three (6%) to mugwort pollen. Furthermore, CJ pollinosis monkeys seemed to be sensitized to these pollen allergens with higher frequency; of 10 monkeys, 10 (100%) had specific IgE to CJ pollen, six (60%) to grass pollen, four (40%) to ragweed pollen, and two (20%) to mugwort pollen.ConclusionJapanese monkeys had specific IgE to grass, ragweed, and mugwort pollen in addition to CJ pollen.
Altered in vitro apoptosis of cultured mast cells prepared from an inbred strain of mice, NC/Kuj
Authors: Hiragun  , Morita  , Shindo  , Tanaka  , Kameyoshi  , Okabe  , Kanno  , Yamamoto  
Published: March 2003

BackgroundAn inbred strain of mice, NC, develops dermatitis associated with highly elevated serum IgE and dermal mast cell hyperplasia.Objectives and methodsTo clarify the mechanisms for the dermal mast cell hyperplasia in NC, we prepared bone marrow‐derived mast cells (BMMCs) from three strains of mice, NC/Kuj, C57BL/6 and BALB/c, and compare histamine contents, histamine release abilities, adhesive properties and apoptosis of the BMMCs.ResultsCompared with BMMCs obtained from C57BL/6 and BALB/c, NC/Kuj BMMC possessed higher histamine content and higher adhesive ability to plastic plates, although histamine release from BMMCs was found to be similar in the three strains. The most intriguing finding is the lack of apoptosis in the BMMCs from NC/Kuj upon growth factor deprivation as determined by DNA ladder formation and loss of membrane phospholipid asymmetry.ConclusionThe altered in vitro properties of mast cells in NC/Kuj partially account for an increase of dermal mast cells, which might be involved in the development of skin lesions in NC.
Association between HLA genes and susceptibility to toluene diisocyanate‐induced asthma
Authors: Mapp  , Beghè  , Balboni  , Zamorani  , Padoan  , Jovine  , Baricordi  , Fabbri  
Published: March 2003

BackgroundOnly a small proportion of subjects exposed to isocyanates develop occupational asthma, suggesting individual predisposition. The human leucocyte antigen (HLA) class II molecules may play a crucial role in the development of the immune response to isocyanates.ObjectivesTo investigate the role of HLA class II molecules in the development of toluene diisocyanate (TDI)‐induced asthma.SubjectsSixty‐seven asthmatic subjects and 27 asymptomatic exposed subjects (controls) were typed at the HLA class II DQA1, DQB1 and DRB1 loci by polymerase chain reaction (PCR)‐based techniques.ResultsThe frequencies of DQA1*0104 and DQB1*0503 were significantly increased in asthmatics compared with asymptomatic exposed subjects, while DQA1*0101 and DQB1*0501 were significantly increased in asymptomatic exposed subjects. No significant difference was found in the distribution of DRB1 alleles between asthmatics and controls.ConclusionsThe results of this study indicate that HLA‐regulated immune mechanisms are involved in TDI‐induced asthma and that, in exposed subjects, specific factors may increase or decrease the risk of developing disease.
Occupational asthma caused by soybean flour in bakers—differences with soybean‐induced epidemic asthma
Authors: Quirce  , Polo  , Figueredo  , González  , Sastre  
Published: March 2003

BackgroundSoybean dust has been identified as the causative agent of occupational asthma and asthma epidemics. Two main soybean hull allergens responsible for asthma outbreaks, Gly m 1 and Gly m 2, have been identified and purified.ObjectiveThe soybean allergens causing occupational asthma in exposed bakers were investigated and compared with those involved in epidemic asthma.MethodsWe report four bakers or confectioners with work‐related respiratory symptoms who were exposed to soybean flour used as a baking additive. The causative role of soybean flour was investigated by immunological tests and specific inhalation challenge tests. Soybean flour allergens causing occupational asthma were characterized by immunoblotting. Immunoglobulin (Ig) E‐reactivity to Gly m 1 and Gly m 2 was assessed using enzyme‐linked immunosorbent assay.ResultsSensitization to soybean flour was demonstrated by skin and serological tests and was confirmed by positive inhalation tests. Bronchial challenge test to soybean flour extract elicited immediate or dual asthmatic responses. Immunoblotting with soybean flour and soybean hull extracts showed IgE‐binding mainly to high molecular weight (MW) allergens. There was an important individually different allergic response to inhalant soybean components. None of the patients showed IgE‐reactivity against Gly m 1 and only one patient showed IgE‐reactivity to the soybean hull allergen Gly m 2.ConclusionThese bakery workers had developed IgE‐mediated occupational asthma to soybean flour. The allergens involved in occupational asthma caused by soybean flour are predominantly high MW proteins that are present both in soybean hull and flour, and they are different from the allergens causing asthma outbreaks, which are mainly low MW proteins concentrated in the hull.
Temporal patterns of mediator release during developing cutaneous late‐phase reactions
Authors: Zweiman  , Von Allmen  
Published: March 2003

BackgroundSeveral inflammatory mediators have been found released in sites of cutaneous late phase reactions (LPR). However, the temporal pattern of their release during LPR development has not been characterized.ObjectiveDetermine hourly accumulation of mediator release in comparison with gross and inflammatory cell responses during developing LPR.MethodsSkin chamber appended to sites of allergen and diluent control challenge with hourly collections. Then, study of exuding leucocytes in chamber bases.ResultsIn the allergen‐challenged sites, histamine release peaked in the first hour, then low level release over the next 5 h. Lactoferrin release from neutrophils started by the second hour, likely associated with released IL‐8. Eosinophil cationic protein levels started increasing slightly later. The percentage of exuding leucocytes which were activated was significantly higher in the allergen challenge sites than in the control challenge sitesConclusionsBoth gross LPR and local inflammatory cell responses in the skin start soon after the immediate mast cell activation in IgE‐mediated responses. Such inflammatory responses include leucocyte activation and mediator release.
Endogenous interleukin‐10 suppresses allergen‐induced airway inflammation and nonspecific airway responsiveness
Authors: Tournoy  , Kips  , Pauwels  
Published: March 2003

BackgroundThe airway inflammation observed in asthma is orchestrated by activated Th‐2 lymphocytes relevant for the induction of altered airway responsiveness. An increasing body of evidence is accumulating that not only the pro‐inflammatory cytokines interleukin (IL)‐4 and IL‐5 but also the immunomodulating cytokines IL‐12 and possibly IL‐10 are crucial for regulating the allergic airway inflammation.ObjectiveSince IL‐10 is capable of downregulating a broad spectrum of pro‐inflammatory cytokines, we wanted to address the role of endogenously produced IL‐10 in vivo in allergic asthma.MethodsKnockout (IL‐10−/−) mice (C57BL/6‐IL10tm1Cgn) and wild‐type (WT) counterparts were immunized (day 0) and exposed (day 14–21) to ovalbumin (OVA). Airway inflammation and reactivity (AR), serum allergen‐specific IgE responses and cytokine profiles in the bronchoalveolar lavage fluid (BALF) were studied.ResultsThe IL‐10−/− mice had more eosinophilic airway inflammation but comparable levels of allergen‐specific serum IgE compared to the WT mice after allergen challenge. The AR was comparably increased in the OVA challenged WT and IL‐10−/− mice vs sham‐exposed WT, but not vs sham‐exposed IL‐10−/− mice since these showed a higher baseline AR. IFN γ, IL‐4 and IL‐13 were comparable and IL‐5 was even lower in the BALF of the in IL‐10−/− mice compared to the similarly exposed WT mice.ConclusionThese results indicate that IL‐10 plays an important and possibly direct role in the control of airway inflammation and responsiveness in an in vivo mouse model of allergy.
Complementary DNA cloning and expression of a newly recognized high molecular mass allergen Phl p 13 from timothy grass pollen (Phleum pratense)
Authors: Suck  , Petersen  , Hagen  , Cromwell  , Becker  , Fiebig  
Published: March 2003

BackgroundGrass pollen extracts contain a range of different allergenic components that can be classified as having low, middle or high molecular mass. Almost 75% of patients allergic to grass pollen display immunoglobulin (Ig) E‐reactivity to allergens in the high molecular mass range of 55–60 kDa. These proteins have not yet been fully characterized on the protein and DNA level.ObjectiveThe aim of this study was to identify and characterize an allergen of the high molecular mass fraction of Phleum pratense pollen by N‐terminal protein sequencing and molecular cloning.MethodsA previously uncharacterized allergen which migrates as a double band with a molecular mass of 55–60 kDa was biochemically purified and investigated by N‐terminal sequencing. Subsequently, a DNA primer was designed to amplify the corresponding cDNA using PCR. The cloned cDNA and deduced amino acid sequence were compared with sequence data bases. Immunoblots carrying the recombinant expression product were developed with monoclonal antibodies and sera derived from allergic subjects. The IgE‐binding capacity of natural and recombinant allergen was determined using EAST.ResultsThe nucleic acid sequence as well as the deduced amino acid sequence consisting of 394 amino acids indicated homology with pollen specific polygalacturonases. Four potential sites for glycosylation and 16 cysteine residues were found. The recombinant expression product exhibited the same molecular size as the natural allergen and was clearly IgE‐reactive.ConclusionThe newly characterized allergen Phl p 13, which shows homology with polygalacturonases, is clearly different from the allergen designated as Phl p 4 and therefore the high molecular mass fraction is composed of at least two different allergens. A possible reason why this important allergen has not been detected until now is that Phl p 13 and Phl p 4 are hardly separable by one dimensional SDS‐PAGE.
Experimental rhinovirus 16 infection increases intercellular adhesion molecule‐1 expression in bronchial epithelium of asthmatics regardless of inhaled steroid treatment
Authors: Grünberg  , Sharon  , Hiltermann  , Brahim  , Dick  , Sterk  , Van Krieken J. Han J. M. 
Published: March 2003

BackgroundRhinovirus infections in airway epithelial cells in vitro have been shown to upregulate intercellular adhesion molecule‐1 (ICAM‐1) expression. Epithelial ICAM‐1, in its dual role as the major rhinovirus receptor and as adhesion molecule for inflammatory cells may be involved in the pathogenesis of rhinovirus‐induced exacerbations of asthma.ObjectiveWe aimed to investigate the effect of experimental rhinovirus 16 (RV16) infection on ICAM‐1 expression in bronchial mucosal biopsies in asthma. In addition, the effect of 2 weeks pretreatment with inhaled budesonide (800 μg b.d.) on RV16‐associated changes in ICAM‐1 expression was studied.MethodsThe study had a parallel, placebo‐controlled design in 25 steroid‐naive nonsmoking atopic asthmatic subjects. After 2 weeks budesonide (BUD) or placebo (PLAC) pretreatment bronchoscopy was performed 2 days before (day −2) and 6 days after (day 6) RV16 inoculation (on days 0 and 1). Immunohistochemical staining for ICAM‐1 was performed on snap‐frozen bronchial biopsies. ICAM‐1 staining intensity on the basal epithelial cells was scored semiquantitatively from 1 (weak) to 3 (intense). Similarly, epithelial intactness was noted (1 = basal cells only, 2 = basal and parabasal cells, 3 = intact epithelium).ResultsICAM‐1 scores were not significantly different between the groups at day −2 (P ≥ 0.08). Subsequent RV16 infection was associated with a trend towards an increase in ICAM‐1 expression in the BUD‐group (P = 0.07), whereas the increase was significant in the PLAC‐group (P = 0.03). However, the increase was not significantly different between the groups (P = 0.74). Epithelial intactness score was not different between the groups before RV16 infection (P ≥ 0.07), and no significant changes were observed in either group (P ≥ 0.59). Moreover, ICAM‐1 score did not correlate significantly with epithelium score in either group, at any time‐point (P ≥ 0.27).ConclusionWe conclude that an RV16 common cold in atopic asthmatic subjects is associated with increased ICAM‐1 expression in the bronchial epithelium, which is not related to epithelial intactness. Glucocorticoid treatment does not appear to prevent the RV16‐associated increased ICAM‐1 expression. This suggests that other treatment modalities are required to protect against the spreading of infection during rhinovirus‐induced exacerbations in asthma.
Nasal hyperosmolar challenge with a dry powder of mannitol in patients with allergic rhinitis. Evidence for epithelial cell involvement
Authors: Koskela H. , Di Sciascio M. B. , Anderson S. D. , Andersson M. , Chan H.‐K. , Gadalla S. , Katelaris C. 
Published: March 2003

BackgroundThe responses to airway hyperosmolar challenges probably involve various inflammatory mediators. However, it is not fully understood which cell type/types are the source of these mediators. Potential cell types include mast cell, epithelial cell and the sensory c‐fibre nerve cell.ObjectiveTo clarify which cell types are involved with the mediator response to hyperosmolarity in the human airway.MethodsTen healthy subjects, 11 patients with nonactive allergic rhinitis, and nine with active allergic rhinitis were challenged intranasally with mannitol powder, and with sham provocation, on separate days. Symptoms were assessed by visual analogue scales and nasal patency by measuring the nasal peak inspiratory flow (nPIF). Nasal lavage fluid levels of α2‐macroglobulin (an index of plasma extravasation), substance P (an index of sensory nerve cell activation), tryptase (an index of mast cell activation) and 15‐hydroxyeicosatetraenoic acid (15–HETE, an index of epithelial cell activation) were analysed.ResultsImmediate, although transient burning was the most prominent symptom in all groups whereas only the patients with active rhinitis experienced a fall in nPIF. Mannitol significantly increased the nasal lavage fluid 15‐HETE levels in the allergic patients (P < 0.01 vs the sham challenge), but not in the healthy subjects. The increase in 15‐HETE correlated with nasal symptoms for itching (rs = 0.65, P = 0.019) and burning (rs = 0.72, P = 0.009). Detectable levels of tryptase was found only in five allergic subjects. Lavage levels of substance P and α2‐macroglobulin did not not change.ConclusionEpithelial cell seems to be involved with the mediator response to airway hyperosmolar challenge. The roles of sensory c‐fibre nerve cell and mast cell remained less clear.