bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased numbers of dendritic cells in the bronchial mucosa of atopic asthmatic patients: downregulation by inhaled corticosteroids
Authors: MÖLLER G. M. , OVERBBEK S. E. , VAN HELDEN‐MEEUWSEN C. G. , VAN HAARST J. M. W. , PRENS E. P. , MULDER P. G. , POSTMA D. S. , HOOGSTEDEN H. C.
Published: April 2006

Background
Dendritic cells (DC) are the most potent antigen‐presenting cells (APC) and stimulators of T cells. Dendritic cells are also likely to be essential for the initiation of allergic immune responses in the lung. However, there are not many data on the presence of dendritic cells in the airways of patients with atopic asthma and on the effects of corticosteroid‐treatment on such dendritic cells.

Objective
We investigated the distribution of dendritic cells in the bronchial epithelium and mucosa of 16 non‐smoking atopic asthmatic patients and eight healthy control subjects using detailed immunohistochemistry (CD l a, HLA‐DR, L25 as markers for dendritic cells).

Methods
Eleven asthmatics were treated for 2.5 years with bronchodilators only and five with bronchodilators and inhaled beclomethasone dipropionate (BDP), 800 μg daily. The patients were randomly sampled from a double‐blind multicentre study. Results There were higher numbers of CD la+ DC (P = 0.003), L25+ DC (P = 0.002) and HLA‐DR expression (P = 0.042) in the bronchial mucosa of asthmatic patients compared with healthy controls. After 2.5 years of treatment, we found a significant increase in fiow expiratory volume in I second (FEV1) (P = 0.009) and a significant decrease in hyperresponsiveness (PC20 histamine) (P= 0.013) in the corticosteroid group (n = 5) compared with the bronchodilator group (n= 11). This dinical improvement in the corticosteroid‐treated group was accompanied by significantly lower numbers of CDla+ DC (P = 0.008), and HLA‐DR expression (P‐ 0.028) in the bronchial mucosa than in the bronchodilator‐treated group.

Conclusion
Our data suggest that dendritic cells are involved in asthmatic inflammation and that corticosteroids may downregulute the number of dendritic.
Function and regulation of FcεRI expression on monocytes from non‐atopic donors
Authors: REISCHL I. G. , CORVAÍA N. , EFFENBERGER F. , WOEFF‐WINISKI B. , KRÖMER E. , MUDDE G. C.
Published: April 2006

Background
The high affinity receptor for IgE (FcεRI) has recently been identified on antigen presenting cells, i.e. Langerhans cells and monocytes from atopic donors and it was hypothesized that FcεRI expression levels correlated with allergy.

Objective
The aims of the study was to investigate the function and expression of FcεRI on monocytes frotn non‐atopic donors.

Methods
Purified monocytes or peripheral blood mononuclear cells were used to study FcεRI expression and signal transduction on CD14 positive cells by flow cytometry and/or confocal laser microscopy.

Results
Freshly isolated monocytes from healthy individuals (n = 58) were shown to express FcεRI (median 18%, range 2–66%). No IgE was bound to these receptors in vivo, and in vitro no significant binding of complete IgE molecules could be obtained. IgE positive monocytes from atopic donors were also found to have free FcεRI, incapable of binding IgE in vitro. Oti all CD14 positive cells free FcεRI expression was rapidly and completely lost during culture in conventiotial culture media (IMDM. RPMI) but not in phosphate buffered saline (PBS). Moreover, signal transduction through free FcεRI appeared to be inhibited. However, both IgE binding and calcium mobilization were restored by treatment of fresh non‐atopic monocytes with neuraminidase. Importantly, culturing these monocytes overnight in conventional medium containing 2μg/mL IgE induced a cycloheximide insensitive accumulation of IgE bound to FcεRI and, in addition, led to cell activation.

Conclusion
Monocytes from both atopic donors and healthy individuals express FcεRI, but the previously reported different expression levels between the two groups seem to be directly related to the absence or presence of TgE in the serum. This may be due to the fact that FcεRI is subjected to a constant turnover process which is slowed down but not prevented by ligand binding. In addition, free FcεRI on non‐atopic monocytes are under control of a neuramindase sensitive structure(s), which influences signal transduction and IgE binding.
Dendritic cells in normal and asthmatic airways: expression of the α subunit of the high affinity immunoglobulin E receptor (FcεRI‐α)
Authors: TUNON‐DE‐LARA J. M. , REDINGTON A. E. , BRADDING P. , CHURCH M. K. , HARTLEY J. A. , SEMPER A. E. , HOLGATE S. T.
Published: April 2006

Background
Immunoglobulin E (IgE) plays an important role in asthma, with total serum IgE levels closely related to both clinical expression of the disease and airway hyperresponsiveness. IgE binds to a high affinity cell‐surface receptor (FcεRI) which is present on mast cells and which has also recently been demonstrated on cutaneous dendritic cells. If pulmonary dendritic cells were also able to express this receptor, this would have important implications with regard to their potential role in asthma.

Objectives
The aims of the study were to investigate the expression of the α subunit of the high affinity IgE receptor (FcεRI‐α) in normal and asthmatic airways, and to analyse its cellular provenance with particular emphasis on the dendritic cell.

Methods
Bronchial biopsy specimens were obtained using fibreoptic bronchoscopy from 10 atopic asthmatics and nine non‐atopic non‐asthmatic control subjects. Specimens were processed into glycolmethacrylate resin and analysed by immunohistochemistry using specific monoclonal antibodies against FcεRI‐α. and against tryptase and CDla. markers for mast cells and dendritic cells, respectively.

Results
The numbers of dendritic cells were significantly higher in the airways of asthmatics compared with those of control subjects (P<0.02). Analysis of sequential sections revealed that the α subunit of FcεRI was localized to both mast eells and dendritic cells. The proportion of dendritic cells expressing FcεRI‐α was significantly inereased in the asthmatic group (P < 0.003).

Conclusion
These results support the hypothesis that dendritic cells play an important role in allergic asthma although the functional significance of FcεRI‐α expression needs further investigation.
Identification of the reactive subunits of Aspergillus umbrosus involved in the antigenic response in farmer's lung
Authors: KAUKONEN K. , PELLINIEMI L. J. , SAVOLAINEN J. , TERHO E. O.
Published: April 2006

Background
Farmer's lung (FL) is the most common form of extrinsic allergic alveolitis and the fungus Aspergillus umbrosus has frequently been associated with FL among Finnish farmers. IgG and IgA class antibodies against Aspergillus umbrosus are found in the serum of FL patients and healthy exposed farmers. So far the immunologically reactive subunits of Aspergillus umbrosus are unknown.

Objectives
The purpose of the study was to identify and localize the reactive subunits of Aspergillus umbrosus that are involved in the antigenic response in FL.

Methods
The distribution of the primary antibody on the outer wall layers of 4‐ and 11 day‐old fungi Aspergillus umbrosus was studied by pre‐embedding immunolabelling with fluorescein isothiocyanate (FITC)‐ and gold‐conjugated antibodies. The surface carbohydrates of the outer wall layers were studied using FITC‐conjugated lectins.

Results
FITC‐conjugated antibodies from FL patient sera reacted with spores (conidia), phialides and hyphae, whereas the fruiting body, cleistothecia, remained unstainable. Immunofluorescence labelling was confirmed by immunolabelling electron microscopy, and gold‐conjugated IgG antibodies were found on the outer cell walls of spinulose conidia, and in a less degree on the outer cell walls of hyphae and phialides. Only single gold particles were found scattered on the surfaces of cleistothecia. The surface carbohydrates of Aspergillus umbrosus conidia. vesicles, phialides and hyphae had a high affinity to GlcNAcβl‐ and Manαl‐/Glcαl‐ specific lectins and only GlcNAcβl‐specific lectins also reacted with the outer cell walls of cleistothecia. Blocking of the antigenic sites of Aspergillus umbrosus by preincubating antigen‐coated enzyme linked immunosorbent assay (FLISA) plates with lectins caused some inhibition, indicating that sites with the highest affinity for lectins were not always antigenic sites.

Conclusion
Gold‐ and FITC‐conjugated antibodies were distributed mostly on the outer wall layers of conidia, hyphae and phialides of Aspergillus umbrosus and GlcN Acβl ‐linked lectins had the strongest affinity to the outer wall layers.
A clinical evaluation in children of the Pharmacia ImmunoCAP system for inhalant allergens
Authors: GLEESON M. , CRIPPS A. W. , HENSLEY M. J. , WLODARCZYK J. H. , HENRY R. L. , CLANCY R. L.
Published: April 2006

Background
The Pharmacia ImmunoCAP system (CAP) for assaying serum IgE specific antibodies was evaluated in a clinical setting against skin‐prick test (SPT) perfonncd using Dome/Hollister‐Steir allergen extracts. The five cotnmon inhalant allergens D. pteronyssinus. D. farinae, mould mix, grass mix and cat epithelium were tested concurrently by both methods in 167 children aged 7.5–12 years. The specific SPT for D. pteronyssinus and D. farinae were also tested against the CAP house dust mite (HDM) mix.

Objective
The purpose of the study was to determine the sensitivity and specificity of the Pharmacia ImmunoCAP system Tor detecting serum IgE specific antibodies to inhalant allergens in a clinical setting, using the SPT result as the ‘gold standard’.

Methods
The SPTs were performed using Dome/Hollister‐Steir allergen extracts. The serum IgE specific antibodies were quantitated using the radioimmunoassay version of the Pharmacia ImmunoCAP system. A history of allergic disease was assessed using a validated questionnaire.

Results
SPT gave more positive reactions than CAP with the exception of cat epithelium. The concordance between SPT and CAP results was 91% over all the tests. The concordance with SPT was slightly higher for the specific CAP for D. pteronyssinus and D. farinae (93% and 95% respectively) than for the CAP HDM mix (86% and 90% respectively). There was a higher proportion of positive results for both SPT and CAP in the 115 children defined as having a history of allergic disease. Using SPT defined allergy as the gold standard, the sensitivity of the CAP system was 87% for the two specific house dust mites but was lower for cat epithelium (67%), mould mix (59%) and grass mix (46%). The sensitivity of the CAP system improved for D. pteronyssinus (96%) and the HDM mix (91%) when tested in subjects defined as having a history of allergy associated disease. The specificity of the CAP system showed less variation between allergens and ranged from 90–99%.

Conclusion
The results of this study of children aged 7.5–12 years demonstrate that, for the inhalent allergens tested, the Pharmacia ImmunoCAP system perfoms well in the setting of known allergic disease.
Further characterization of histamine releasing chemokines present in fractionated supernatants derived from human mononuclear cells
Authors: KUNA P. , REDDIGARI S. R. , RUCINSKI D. , KAPLAN A. P.
Published: April 2006

Background
Histamine releasing factors (HRF) are members of the β chemokine family of cytokines and have been characterized using recombinant proteins. Mononuelcar cell and/or platelet supernatants have been shown to contain HRF and the initial void peak obtained using Mono Q anion exchange chromatography possesses such activity, as do two later peaks eluted from the column.

Objective
We wished to further characterize the activity present in the void peak and determine which of the chemokines present are responsible for the activity measured.

Methods
We fractionated the void peak obtained from Mono Q chromatography on Mono S. The elution profile of individual chemokities was determined and the fractions were assayed for histamine releasing capability. We used monospecific antisera to inhibit the activity and quantitate the contribution of each protein.

Results
The fractions contained MCP‐1/MCAF, CTAPIII/NAP‐2, IL8, and a small quantity of RANTES. About 90‐95% of the total histamine containing capability was attributable to MCP‐1/MCAF. There was a small contribution by CTAPIII/NAP‐2, and RANTES, and no activity associated with IL8.

Conclusion
MCP‐1/MCAF is the critical HRF present in the initial void peak obtained by anion exchange chromatography of supernatants derived from human mononuclear cells and platelets. The α chemokine CTAPIII/NAP‐2 has relatively weak activity and IL8 has none although they are prominent in this fraction and overlap with MCP‐1/MCAF. RANTES makes a minor contribution but most of it is eluted in a later peak.
Comparative degree and type of sensitization to common indoor and outdoor allergens in subjects with allergic rhinitis and/or asthma
Authors: BOULET L.‐P. , TURCOTTE H. , LAPRISE C. , LAVERTU C. , BEDARD P.‐M. , LAVOIE A. , HÉBERT J.
Published: April 2006

Background and objectives The determinants of variability in the clinical expression of atopy are still to be documented. The goals of this study were to determine, in subjects with a clinical diagnosis of symptomatic asthma or rhinitis, what is the possible contribution of different types of indoor and outdoor allergens to the development of their disease, by looking at the prevalence and degree of sensitization to these allergens according to age and gender.

Subjects and methods We analysed allergy skin prick tests to common airborne indoor and outdoor allergens in 3371 consecutive patients, grouped according to diagnosis of allergic asthma, rhinitis, or both. For each of these three groups, we calculated the prevalence of sensitization to indoor/outdoor allergens, the atopic index (Al), the number of positive responses to allergy skin prick test and the mean wheal diameter (MWD) of these responses.

Results The prevalence of atopy and the values of Al and MWD peaked in subjects aged 16 to 25 years, declining afterwards; in subjects ± 18 years old, atopic indices were slightly higher in men than in women. In atopic subjects, the prevalence of sensitization was, in decreasing order: housedust (84.2%), cat hair‐epithelium (76.5%), dog hair‐dander (63.0%), house dust mite (54.2%), grasses (51.9%), trees (47.2%) and ragweed pollens (44.9%) and finally, moulds (25.4%). Among subjects sensitized only to outdoor allergens (n= 195), 73.8% had a rhinitis, 11.8% had asthma and 14.4% had both diagnoses; for those sensitized only to indoor allergens (n= 710), these values were respeetively 48.6, 24.5 and 26.9%, and for those sensitized to both indoor and outdoor allergens (n= 1793), the comparable values were 55.5, 14.6 and 29.9%.

Conclusion These data show that in our population of subjects with respiratory allergic symptoms, indoor allergen sensitization is strongly associated with asthma, while exclusive sensitization to pollens is associated primarily with rhinitis. Sensitization was more prevalent for indoor allergens than for outdoor allergens in all groups determined according to diagnosis or age. Indices of atopy were higher in men in the group ± 18 years old. Prevalence and degree of sensitization were shown to peak in young adults, regardless of the allergen, and to diminish with age. This study stresses the role of indoor allergens in the development of asthma and shows the variability of allergic manifestations according to the type of sensitization.
Measurement of IgE antibodies against purified grass pollen allergens (Lol p 1, 2, 3 and 5) during immunotherapy
Authors: VAN REE R. , VAN LEEUWEN W. A. , DIEGES P. H. , VAN WUK R. GERTH , DE JONG N. , BREWCZYSKI P. Z. , KROON A. M. , SCHILTE P. P. M. , TAN K. Y. , SIMON‐LICHT I. F. , ROBERTS A. M. , STAPEL S. O. , AALBERSE R. C.
Published: April 2006

Background IgE titres tend to rise early after the start of immunotherapy, followed by a decline to pre‐immunotherapy levels or lower.

Objectives We were interested to ktiow whether the early increase in IgE antibodies includes new specificities of IgE, and whether these responses persist.

Methods Sera of 64 patients undergoing grass pollen immunotherapy were tested for IgE against four purified grass pollen allergens: Lol p 1. 2, 3, and 5. At least two serum samples were taken, one before the start of therapy and one between 5 and 18 months after the first immunization (mean: 10 months).

Results The mean IgE responses to Lol p 1, 2 and 3 showed a moderate but not significant increase. In contrast, the mean IgE response to Lol p 5 showed a significant decrease of >30%. IgE against total Lolium perenne pollen extract moderately increased (>20%), showing that a RAST for total pollen is not always indicative for the development of IgE against its major allergens. For >40% of the patients it was found that IgE against one or more of the four allergens increased, while IgE against the remaining allergen(s) decreased. Eor 10 sera the ratio of IgE titres against at least two allergens changed by at least a factor of 5. The changes in specific IgE also included conversions from negative (< 0.1 RU) to positive (0.6 to 5.0 RU) for five patients. For two patients, the induction of these ‘new’ IgE antibodies against major allergens was shown to result in a response that was persistent over several years.

Conclusion Although active induction of new IgE specificities by immunotherapy was not really proven, the observations in this study indicate that monitoring of IgE against purified (major) allergens is necessary to evaluate changes in specific IgE in a reliable way.
Oilseed rape pollen is a potentially relevant allergen
Authors: HEMMER W. , FOCKE M. , WANTKE E. , JÅGER S. , GÖTZ M. , JARISCH R.
Published: April 2006

Oilseed rape (Brassica napus) (OSR) is a partly wind‐pollinaled crop which has been increasingly cultivated both in Europe and overseas. Allergological data about OSR is scarce and controversial. We evaluated the frequency of sensitizalion to OSR pollen by skin prick test and RAST over a period of 1 yr. Airborne OSR pollen load and the agricullural role of this crop were analysed. Furthermore, six patients were investigated by immunoblot. In 4468 patients with suspect inhalant allergy investigated between June 1994 and May 1995, routine skin prick testing revealed OSR sensitivity in 7.1% ot pollen‐allergic patients. In all, monovalent sensitization was detected in nine patients. Routine pollen counts showed daily maxima not exceeding 50 grains/m3/24 h, but airborne OSR pollen has continuously increased during the last decade correlating with the increasing acreage. Characterization of OSR allergens by immunoblol revealed major allergens of 6/8 kD, 12/14 kD and in the high molecular weight range at 33, 42, 51, 58/61 and 70kD. Some OSR proteins may cross‐react with birch pollen allergens. In summary, the results suggest that OSR pollen is a moderate but true source of allergy and may sensitize despite low pollen exposure.
Characterization of allergens from deer: cross‐reactivity with allergens from cow dander
Authors: SPITZAUER S. , VALENTA R. , MÜHL S. , RUMPOLD H. , EBNER H. , EBNER C.
Published: April 2006

Background Animal hair/dander proteins frequently cause Type I hypersensitivities. Species‐specific and broadly cross‐reacting allergens have been characterized in the past.

Methods Sera from eight individuals suffering from symptoms due to exposure to deer and deer‐derived products were investigated by immunoblotting. Extracts from deer, dog, cat, horse, rabbit and cow, respectively, were tested for IgE‐binding. To reveal cross‐reactivities patients' sera were preadsorbed with these extracts prior to testing with deer extract.

Results Deer allergens with the molecular mass of 22 and 25 kD (major allergens), as well as 60 kD were identified. The 22 and 25 kD allergens are cross‐reactive with the corresponding cow allergens.

Conclusions Deer allergy is a rare sensitization mainly affecting persons exposed to deer, who displayed an atopie disposition. From our results it can be assumed that this hypersensitivity is partly associated with allergy to cow dander.
The measurement of reticular basement membrane and submucosal collagen in the asthmatic airway
Authors: WILSON J. W. , LI X.
Published: April 2006

Background An increase in types III and V collagen below the bronchial epilhelium of the airway is known to be responsible for apparent thickening of the reticular basement membrane (RBM) and is recognized as a characteristic feature of asthma. Wide variations exist in reported ranges of the thickness of the RBM in both asthmatic and control airways. The description of collagen deposition has been confined to the RBM with no previous reports of increased collagen deeper within the bronchial wall (i.e. submucosa), although inflammation is known to extend to deeper levels.

Objective The objective of this study was to compare different methods used to measure RBM thickness and to examine collagen deposition in deeper asthtnatic bronchial submucosa.

Methods We examined bronchial mucosal biopsies from 15 asthmatic and 13 normal healthy controls to measure RBM thickness by light and electron microscopic methods on resin sections and to estimate the extent of collagen deposition in the submucosa by immunochemical staining with monoclonal antibodies to collagen subtypes on paraffin sections.

Resutts Simple staining with toluidine blue on resin sections coupled with light microscopic evaluation was as accurate and less expensive than electron microscopic methods in the measurement of the thickness of RBM (mean ± sd, light tnicroscopy: asthma vs control. 6.01 ± 2.09 vs 3.19 ± 0.55 μm,P < 0.001; electron microscopy: asthma vs control. 6.09 ± 2.19 vs 2.85 ± 0.86 μm, P < 0.01). Using monoclonal antibodies to collagen subtypes and image analysis methods, we identified increased collagen in the submucosa in asthma (type III collagen, asthma vs control: 62 ± 7 vs 51 ± 9%,P < 0.05, type V collagen, asthma vs control: 60 ± 6 vs 52 ± 9%, P < 0.05).

Conctusions This study has demonstrated that the RBM may be reliably measured by light microscopic methods. This study shows for the first time that the submucosa of the asthmatic airway contains significantly more collagen than nonnal controls, giving rise to the possibility that airway scar formation may have greater functional implications than has been previously believed from consideration of the RBM alone.
Purification and characterization of a soybean hull allergen responsible for the Barcelona asthma outbreaks. II. Purification and sequencing of the Gly m 2 allergen
Authors: CODINA R. , LOCKEY R. F. , EERNÁNDEZ‐CALDAS E. , RAMA R.
Published: April 2006

Background A low MW allergen from soybean hull, Gly m 1, with two isoallergens, Gly m 1 A and Gly m 1 B, was associated with the asthma outbreaks that occurred in Cartagena, Spain. Using sera of asthmatic epidemic patients (AEP) from Barcelona, three main soybean hull allergens, two of them with MWs atid pIs identical to those reported for Gly m 1 A and Gly m 1 B, were identified.

Objective The purpose of this study was to purify and to study the N‐terminal amino acid sequence of the third allergen, which has a MW of 8 kDa.

Method The purification procedure combined the double dialysis method and preparative isoelectofocusing (IEF). Specific IgE determination to the fractions obtained demonstrated three peaks, one of them corresponding to the 8 kDa allergen. The pooled fractions containing this allergen were studied by sodium dodecylsulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE), SDS‐PAGEAVestern blot and IEF/Westem blot. Only a band with a MW of 8 kDa and a pI of 6 was obtained. Its allergenic activity was measured and it was demonstrated that the allergenicity of soybean hull correlates with the presence of the 8 kDa allergen. The N‐terminal amino acid sequence of the first 20 amino acids, which was registered at the PIR Data Submission as the N‐terminal partial sequence of Gly m 2, was determined according the Edman degradation method.

Results Gly m 2 N‐terminal amino acid sequence lacks homology with that reported for the allergen Gly m 1 but has a homology of 71% with a storage protein from cotyledon of Vigna radiata (cow pea) and 64% with a‘disease response protein’ from Pisum sativum (green pea). These results suggest that Gly m 2 in soybeans could protect against diseases which affect soybean plants.

Conclusion This study demonstrates the existence of another soybean hull allergen, Gly m 2, partially responsible for the soybean asthma outbreaks that occurred in Barcelona, Spain.
Hay fever, asthma and number of older siblings — a twin study
Authors: RäSäNEN M. , LAITINEN T. , KAPRIO J. , KOSKENVUO M. , LAITINEN L. A.
Published: April 2006

Background It has been suggested that allergic sensitization is inversely related to the number of siblings in the family.

Objectives To study whether a similar relation can be observed for hay fever and asthma among Finnish adolescents in a population with relatively low prevalence of atopic diseases.

Methods A questionnaire mailed to a nationwide sample of 1849 families with 16‐year‐ old twins assessing the cumulative incidence of doctor‐diagnosed hay fever and asthma among the adolescents and the number of older siblings in the family by parental report.

Results The cumulative incidence of hay fever was significantly lower among the adolescents with three or more older siblings (3.9%, 95% CI= 1.2–6.5%) compared with adolescents with fewer older siblings (12.7%, 95% CI=11.4–14.0%). There was no difference in the cumulative incidence of asthma among the adolescents according to the number of older siblings in the family.

Conclusions Large number of older siblings appears to be protective against the development of hay fever.
Asthma, bronchial hyperreactivity and mediator release in children with birch pollinosis. ECP and EPX levels are not related to bronchial hyperreactivity
Authors: FERDOUSI H. A. , DREBORG S.
Published: April 2006

Background Symptoms of allergic asthma are triggered by allergen exposure inducing allergic inflammation and hyperreactivity of the bronchi.

Objectives To investigate the possible relationship between clinical symptoms and signs of asthma, i.e. bronchial variability as measured by peak expiatory flow rate (PEFR). bronchial hyperreactivity (BHR) and mediators of allergic inflammation.

Methods Twenty‐eight children with pollinosis. but no obvious history of asthma, were studied at three occasions, i.e. before, during and after (autumn) the birch pollen season. Twelve children sensitive to birch pollen were considered as the case group. Sixteen children, who were only clinically sensitive to grass pollen, served as controls. Subjective symptoms of asthma were recorded by visual analogue scale, BHR was estimated by methacholine bronchial provocation tests, bronchial variability PEFR and circulating mediators of inflammation, i.e. eosinophil cationic protein, eosinophil protein X, myeloperoxidase and tryptase in serum.

Results Bronchial hyperreactivity and by PEFR was more pronounced after than during the season (P < 0.01), whereas eosinophil mediators and the peak expiratory flow rate increased during the season (P < 0.05). Except for between PEFR variability and BHR in the autumn (r= 0.45; P= 0.014). no correlations were found. However, in the autumn, the majority of children were still hyperreactive in the bronchi and showed PEFR variability but the levels of eosinophil mediators in serum had returned to normal levels.

Conclusion Signs and symptoms of asthma did not correlate with serum levels of mediators of allergic inflammation. Bronchial hyperreactivity and PEFR variability persisted after the pollen season when signs of bronchial inflammation had disappeared. We hypothesize that eosinophil mediators and other markers of allergic inflammation disappear after the late‐phase reaction, whereas BHR persists. This would explain the lack of correlation between the levels of eosinophil mediators in serum and symptoms of asthma and BHR.
The interaction of tumour necrosis factor alpha and endothelin‐1 in pathogenetic models of asthma
Authors: XU J. , ZHONG* N. S.
Published: April 2006

Background There is evidence that tumour necrosis factor alpha (TNFα) may be an important mediator in initiating asthmatic airway inflammation. It has been proposed that endothelin‐1 (ET‐1) is involved in bronchoconstriction and airway remodelling in asthma. It is not known, however, if there is any interaction between TNFα and ET in perpetuating airway inflammation in asthma.

Objective The present study aimed to determine the activities of ET‐1 and TNFα in ovalbumin‐sensitized guinea pigs and their roles in the development of airway inflammation.

Method Twelve guinea pigs were sensitized by ovalbumin injection and aerosol inhalation. ET‐1 levels were measured in both bronchoalveolar lavage fluid (BALF) and plasma by 125‐labelled endothelin‐1 (ET‐1) radioimmunoassay. The TNFα activity released from alveolar macrophage (AM) in BALF was estimated by ELISA. Cultured bovine airway smooth muscle cells (BASMCs) were treated with TNFα (1000 units/5 ± 104 cells) for different times. ET‐1 levels in harvested medium from these cells were measured by radioimmunoassay. Cultured human fetal lung fibroblasts (HFLFs) were incubated with ET‐1(10–8–10–6M), then 3HTdR incorporation to these cells and cell counting were performed. The effects of ET‐1 stimulation on the granulocyte macrophage colony stimulating factor (GM CSF) gene expression in HFLFs were estimated by using RT‐ PCR method.

Results ET‐1 levels in both plasma and BALF were significantly higher in ovalbumin‐ sensitized guinea‐pigs compared with those in controls (422.27 × 175.0pg/mL vs 277.311 × 88.0pg/mL, P < 0.05, 81.22 × 16.15 vs 49.81 × 12.64pg/mL, P < 0.05) while TNFα activity was also significantly increased in the OVA‐sensitized group compared with that in the control group (6010 ± 1900pg/mL vs 2810 × 450 pg/mL, P < 0.05). The ET‐1 level in harvested medium of BASMCs rose significantly in 12 h in the TNF‐α treated group (from < 5pg/mL to53.72 × 14.3pg/mL, P < 0.001), and remained at a similar level for 24 h in the TNFα treated group. It was shown that ET‐1 not only stimulated cell proliferation but also induced GM‐CSF mRNA expression in HFLFs.

Conclusion ET‐1 levels in both plasma and BALF and TNFα release from macrophage are increased significantly in ovalbumin‐sensitized guinea‐pigs. TNFα stimulates ET‐1 secretion from cultured BASMCsw; ET‐1 accelerates cell proliferation and induces GM‐ CSF mRNA expression in the human fetal lung fibroblast.
Sensitivity to bee and wasp venoms: association with specific IgE responses to the bee and wasp venom and HLA DRB1 and DPB1
Authors: FAUX J. A. , MOFFATT M. F. , LALVANI A. , DEKKER J. , WARRELL D. A. , COOKSON W. O. C.
Published: April 2006

Background Stings from bees and wasps can cause systemic reactions which can be fatal in some individuals. In these venom‐sensitive patients, specific IgE to the venom is produced and is considered to participate in the adverse reactions. This immune response requires antigen presentation by human leucocyte antigens (HLA) class II molecules, which includes DR and DP, which are present on antigen presenting cells.

Objective To test for associations between HLA class II DRB1 and DPB1 alleles and life‐ threatening sensitivity to both bee and wasp venoms. To establish further whether any associations are independent of the atopy phenotype.

Methods A total of 33 bee‐ and 44 wasp‐venom‐sensitive patients was studied. DRB1 genotypes were determined by single stranded oligonucleotide (SSO) probing of PCR products, and DPB1 genotypes by amplified fragment length polymorphism (AFLP) analysis. Total and specific IgE were measured using the Pharmacia Immunocap, FEIA. Patients with specific IgE to the venom antigens only were termed monosensitive and those with additional specific IgE to HDM and/or GP were termed polysensitive.

Results Allele frequencies were compared to an unrelated control population. The 33 bee‐ sensitive patients had a greater prevalence of DRB 1*07 alleles than the control subjects, 26% vs 14%, with an odds ratio (OR) of 2.1 (95%CI, 1.2–3.7, P= 0.015, corrected for multiple comparisons, pc > = ns). This association was confined to the 15 monosensitive bee patients, who had a 43% DRB 1*07 allele frequency when compared with 11% in the 18 polysensitive bee patients, OR 6.1 (95%CI, 1.73–22, P= 0.004, pc= 0.05), and when compared with a control group of non‐venom subjects, 43% vs 16%, OR 3.9 (95% CI, 1.72–9.0, P= 0.002, pc= 0.02). The 44 wasp‐sensitive patients had an increase in the DRB1*11 allele when compared with the control subjects, 13% vs 6%, with an OR 2.2 (95%CI, 1.0–4.6, P= 0.04, pc= NS), and a decreased prevalence of DRB 1*04 alleles, 10% vs 19%, with an OR 0.33 (95%CI, 0.24–0.99, P= 0.04, pc= NS), but these were not significant when multiple comparisons were taken into account. The DPB1 alleles were not significantly different between the venom sensitive patients and the controls.

Conclusion Patients monosensitive to bee venom had a significantly greater prevalence of DRB 1 *07 alleles than the non‐venom, control population suggesting that IgE responses in these patients may, in part be controlled by immune response HLA class II genes. These results are also suggestive of wasp‐sensitive patients having a higher prevalence of DRB1*11 and a lower prevalence of DRB 1*04 than the control population.
Cell surface expression of two major yeast allergens in the Pityrosporum genus
Authors: ZARGARI A. , EMILSON A. , HALLDÉN G. , JOHANSSON S. , SCHEYNIUS A.
Published: April 2006

Background We have previously identified two major allergens of Pityrosporum orbiculare and characterized these as 37 kDa and 67 kDa proteins.

Objective In the present study we have investigated the presence and subcellular location of the 37 kDa and 67 kDa allergen components in various members of the genus Pityrosporum as well as in Candida albicans. Candida parapsilosis and Saccharomyces cerevisiae. Methods To detect both cell surface and intracellular expression of the allergens, flow cytometry and confocal laser scanning microscopy (CLSM) were used. The cells were stained with indirect immunofluorescent (IIP) or alkaline phosphatase antialkaline phosphatase (APAAP) methods using mouse monoclonal antibodies (MoAbs)

Results Ninety‐five per cent of the P. orbiculare (P. ovale) cells cultured for 4 days showed cell surface‐binding of the anti‐37 kDa MoAb and 88% of the cells bound the anti‐ 67 kDa MoAb when analysed with IIF and flow cytometry. It was found that the members of the genus Pityrosporum (Malassezia). P. pachydermatis and M. sympodialis, expressed the 37 kDa and 67 kDa allergens to a similar extent as did P. orbiculare. Less than 5% of the cells of the genus Candida and S. cerevisiae showed positive staining with the MoAbs, The CLSM revealed that the 37 kDa and the 67 kDa components were located to the cell wall and could not be detected inside the acetone fixed and APAAP stained yeast cells of the genus Pityrosporum. When the yeast cells were cultured for more than 4 days the expression of both allergens decreased significantly.

Conclusion All three members of the genus Pityrosporum express the 37 kDa and 67 kDa major allergens on the cell surface, whereas these proteins could virtually not be detected in the Candida genus and S. cerevisiae.
Wheat flour peroxidase is a prominent allergen associated with baker's asthma
Authors: SÁNCHEZ‐MONGE R. , GARCÍA‐CASADO G. , LÓPEZ‐OTÍN† C. , ARMENTIA A. , SALCEDO G.
Published: April 2006

Background
Putative wheat allergens with molecular sizes around 35kDa have been repeatedly detected in immunoblots using sera from patients with baker's asthma. However, none of these allergens had been previously isolated.

Objective
To purify and characterize a major component of the 35 kDa aliergenic band which seems to be present in flour from diploid. tetraploid (pasta) and hexaploid (bread) wheats.

Methods
Gel‐filtration chromatography followed by RP‐HPLC have allowed the purification of a new wheat allergen. Immunodetection with sera from allergic patients and with a specific serum for asparagine‐linked complex glycans, as well as amino acid sequencing, were used to identify the isolated protein.

Results
A 36 kDa allergen has been purified from wheat flour. Its N‐terminal and internal sequences, N‐linked glycans, and enzymatic activity indicated that this allergen is a seed‐specific peroxidase. Sera from six out of 10 patients hypersensitive to wheat flour displayed positive reactions when assayed against the purified dot‐blotted allergen.

Conclusion
Seed‐specific peroxidases from cereals (flours) seem to be important allergens in hypersensitive diseases associated with the manipulation of these plant materials. A new enzyme has thus been involved in IgE‐mediated diseases.
Induced sputum to assess airway inflammation: a study of reproducibility
Authors: SPANEVELLO A. , MIGLIORI G. B. , SHARARA A. , BALLARDlNI L. , BRIDGE P. , PISATT P. , NERI M. , IND P. W.
Published: April 2006

Background
Infiltration of the airways mucosa with activated inflammatory cells appears to be a major factor in the pathogenesis of asthma and other airway diseases. Examination of sputum provides a direct method to investigate airway inflammation non‐invasively.

Objectives
The aim of the present study was to evaluate the reproducibility of cell counts on cytospins and fluid phase (eosinophil cationic protein, ECP) measurements in a selected portion of induced sputum. We aimed to confirm the validity of the tecnique by comparing measurements between stable asthmatics, allergic rhinithis and healthy subjects.

Methods
Sputum was induced with hypertonic saline (4.5%) twice within one week in 53 stable asthmatics, 16 subjects with seasonal rhinitis (out of the pollen season), and 19 healthy subjects. Reproducibility was examined within sample (two different plugs of the same sample) between sample (two specimens of induced sputum obtained within one week) and between examiners on stable subjects taking into account sample size, number of examinations per patients and Confidence Interval (CI) of the estimates.

Results
We have found that the method is highly reproducible within sample and between examiners for all types of cells and fluid phase measurements of ECP. It is reproducible between sample for eosinophils, macrophages, neutrophils and ECP, but not for lymphocytes and weakly for epithelial cells. Sputum from asthmatics, in comparison with the sputum of healthy subjects and subjects with rhinitis had higher eosinophils (asthmatics: 12.2%± 12.9, rhinitis: 0.4 ± 0.8, normals: 0.4 ± 0.7(%) and ECP (asthmatics: 827 ± 491 μg/L, rhinitis: 127 ± 82 normals: 157 ± 203). No significant differences were found between healthy subjects and subjects with rhinitis. Eosinophil counts were inversely correlated with FEV1 (r=−0.37) expressed as percentage of predicted, but not significantly correlated with PC20 methacholine (r =−0.28) or blood eosinophils (r = 0.26).

Conclusions
The importance of this study is the confirmation, within important statistical guidelines for a study of reproducibility, that the methods examined are reproducible and valid.
Loratadine treatment of rhinitis due to pollen allergy reduces epithelial ICAM‐1 expression
Authors: CIPRANDI G. , PRONZATO C. , RICCA V. , PASSALACQUA G. , DANZIG M. , CANONICA G. W.
Published: April 2006

Background
Loratadine and cetirizine are new generation antihistamines, which are clinically effective in the treatment of allergic rhinitis.

Objective
The aim of the study was to evaluate antiallergic activity of loratadine compared with cetirizine, over a 2 week period under natural allergen exposure, in a double‐blind parallel groups, randomized, controlled trial.

Methods
Twenty patients, sensitized to grass and/or Parietaria pollen, were subdivided into two groups, one receiving loratadine the other cetirizine respectively. Both were dosed at 10 mg/day. Evaluated parameters were: clinical symptoms, nasal inflammatory cell (such as neutrophil, eosinophil and metachromatic cells) counts, ICAM‐1 expression on nasal epithelial cells, and nasal mediators (e.g. histamine, ECP, EPO and MPO).

Results
Loratadine and cetirizine significantly improved symptoms (P < 0.002), significantly reduced eosinophil (P < 0.016) and metachromatic cell (P < 0.01) infiltration, levels of ECP (F < 0.002), EPO (P < 0.006) and histamine (P < 0.01) and ICAM‐1 expression on nasal epithelial cells (P < 0.02). No difference was demonstrated between the two drugs.

Conclusion
The antiallergic activity of loratadine and cetirizine is documented by their actions on the inflammatory and clinical parameters, especially ICAM‐1 modulation.
Malnutrition and experimental lung allergy
Authors: CUNHA M. G. , NASPITZ C. K. , MACEDO‐SOARES F. , AMÂNCIO O. M. , JANCAR S.
Published: April 2006

Background
Although it is commonly accepted that the immune response is affected by malnutrition there are very few data about its effect in allergie diseases.

Objective
The aim of the present study was to investigate the effect of malnutrition in allergic lung inflammation.

Methods
An anaphylactic reaction was induced in rat lungs and the increased vascular permeability was measured in the trachea, internal and external bronchi and parenchyma by the Evans blue extravasation method. These studies were conducted in two dietary groups: one fed a normoproteic diet (18%) and the other a hypoproteie diet (4.5%). When the animals were 60 days old the group fed the hypoproteie diet presented a reduction of 77.86% in bodyweight, 63.3% in food intake and 36% in plasma protein concentration characterizing a severe protein‐calorie malnutrition.

Results
The anaphylactic reaction in the lungs induced a significant increase in vascular permeability in the trachea and bronchi of both dietary groups. However, the intensity of this effect was significantly lower in the malnourished group. Analysis of immunoglobulin isotypes in the serum by ELISA showed that whereas IgG1 and IgG2a levels were similar in both groups, the levels of IgE were significantly lower in the malnourished animals. Moreover, the levels of antigen‐specific IgG1, IgG2a and IgE were all significantly inhibited by the protein‐calorie malnutrition. When antibodies were passively transfered to the malnourished rats, they developed a reaction as intense as the normoproteic group.

Conclusions
These results suggest that the capacity to release infiammatory mediators and the vascular response to these mediators is not affected by this type of malnutrition and, therefore, the diminished response of the airways reported here is probably due to the lower levels of anaphylactie antibodies produeed by the malnourished rats.
Wasp venom immunotherapy changes IgG antibody specificity
Authors: MICHILS A. , LEDENT C. , MAIRESSE M. , GOSSART B. , DUCHATEAU J.
Published: April 2006

Background
The evolution of the IgG response during venom immunotherapy (VIT) has been previously investigated in terms of antibody titres and subclasses.

Objectives
The present work studied the evolution of IgG antibody fine specificity in wasp allergic patients treated with rush VIT.

Methods
Antibody specificity was evaluated in 51 wasp allergic patients in competitive ELISA using streptavidin biotin technology. Patients were tested before and during specific rush immunotherapy (at 15 days, 6 months, 12 months) and compared with 44 patients treated by venom injections for at least 2 years.

Results
The capacity of sera to prevent the antigen binding of pooled IgG from allergic patients changed rapidly with mean percentage inhibitions (± sd) falling from 70 ± 11–51 ± 18% after 15 days of treatment (P < 0.001 by one way anova). Similarly, the antigen binding capacity of pooled IgG from VIT patients was differently prevented by sera with mean percentage inhibitions increasing from 37 ± 12–65 ± 8 after 15 days of treatment (P < 0.0001 by one‐way anova).

Conclusions
The immunodominance pattern of IgG epitopes recognized on wasp venom antigens by sera from wasp allergic patients changes soon after initiating rush VIT. Further studies will indicate whether, instead of measuring IgG titres, this marked change could be used as the basis of a new test for monitoring the outcome of VIT.
Bronchial and cutaneous responses in atopic dermatitis patients after allergen inhalation challenge
Authors: BRINKMAN L. , ASLANDER M. M. , RAAIJMAKERS J. A. M. , LAMMERS J.‐W. J. , KOENDERMAN L. , BRUIJNZEEL‐KOOMEN C. A. F. M.
Published: April 2006

Background
Atopic dermatitis (AD) is often associated with allergic asthma (AA). Inhalation of allergens influences the activity of AA but the effect on the skin in AD is unclear.

Objectives
We evaluated the degree of bronchial hyperresponsiveness to methacholine in eight AD patients with AA (AD+) and eight AD patients without AA (AD–) and studied bronchial and cutaneous responses after allergen inhalation challenge.

Methods
All patients were treated in hospital for their eczema with tar ointment (pix liquida) and orally administered antihistamines (mean hospital stay 37 days). After clearing of the skin lesions allergen inhalation challenge was performed. Cutaneous responses were studied by measuring the‘Costa’ score before and 24 h after allergen inhalation challenge.

Results
The median value of the provocative concentration of methacholine causing a 20% fall (PC20 Mch) in forced expiratory volume in 1 second (FEV1) was significantly higher in the AD– group compared to the AD+ group with median values of 10.70 and 0.60mg/mL, respectively. These values did not change significantly in both groups during hospital stay. After challenge all AD+ patients showed early and late asthmatic responses whereas only four AD– patients showed early asthmatic responses (mean values of the maximal fall in FEVl during the EAR 37%/16% and in PEF during the LAR 27%/4% for AD+ and AD–patients, respectively). The‘Costa’ score increased in both groups (mean score before 19.1/ 24.4 and after challenge 26.8/26.9 for AD+ and AD+ patients, respectively). The increase in the AD– group was significantly higher compared with the AD– group (P= 0.016).

Conclusion
We conclude that allergen inhalation challenge causes a flare up of the skin lesions in atopic dermatitis patients. This was more prominent in atopic dermatitis patients who already suffered from an IgE‐mediated allergic inflammation in the lung.