bluuebunny/crossref_metadata_2025 huggingface dataset:
Sources of Job and Worker Flows: Evidence from a Panel of Regions
Authors: Böckerman Petri , Hämäläinen Kari , Maliranta Mika
Published: March 2004

Abstract.  This study explores the structure and the dynamics of regional job and worker flows. The measures of job and worker flows are related to economic fluctuations, demographic factors and industry structure by employing the data of 85 Finnish regions over the period 1988–97. It is shown that labour market dynamics differ markedly between regions. As in previous analyses of linked employer–employee data, job and worker flows are shown to behave cyclically. In addition, the results indicate that observable differences in regional productivity, in‐migration, demographics and industry structure help to explain the prevailing disparities in regional labour markets.
Is the Swiss Labor Market Segmented? An Analysis Using Alternative Approaches
Authors: Sousa‐Poza Alfonso
Published: March 2004

Abstract.  In this paper, three common empirical methods encountered in the segmentation literature are used in order to establish whether or not the Swiss labor market is segmented: (i) a hierarchical cluster analysis; (ii) a switching model with unknown regime; and (iii) an analysis of low‐wage mobility with a bivariate probit model with endogenous selection. According to method (i), segmentation can hardly be observed. Method (ii) shows that the Swiss labor market is dualistic in nature. Method (iii) reveals that a certain degree of persistence exists in low‐wage jobs. Whether or not the Swiss labor market is segmented thus depends on the choice of method, i.e. on the definition and understanding of segments. In any case, none of the methods used in this study point to the existence of a large and well‐defined secondary segment.
Increased expression of allergen‐induced in vitro interleukin‐10 and interleukin‐18 mRNA in peripheral blood mononuclear cells of allergic rhinitis patients after specific immunotherapy
Authors: Savolainen J. , Laaksonen K. , Rantio‐Lehtimäki A. , Terho E.O.
Published: March 2004

Background
During specific pollen immunotherapy (SIT) there is a local mucosal shift from Th2‐ to Th1‐ type cytokine predominance, with IL‐12 having a major role in this shift. IL‐10‐induced tolerance is supposed to be a key phenomenon in venom immunotherapy (VIT). However, the role of Th1‐promoting cytokines, on the one hand, and the role of regulatory cytokines, on the other hand, have not been studied in parallel during SIT.

Objective
This study was undertaken to analyse the allergen‐induced in vitro mRNA expression of Th1‐type effector cytokine IL‐18 and regulatory cytokines IL‐10 and TGF‐β during SIT in peripheral blood mononuclear cells (PBMC) of allergic rhinitis (AR) patients.

Methods
Thirty patients with AR undergoing pollen SIT and 10 patients with AR who were not treated with SIT were included in the study. The symptoms and medications were registered post‐seasonally before the beginning of SIT and after 1 year of therapy. PBMC samples were collected and stimulated with pollen allergen extract prior to the treatment, at the maintenance phase in 12 patients and after 1 year of the treatment. The cytokine mRNA expression was assessed using kinetic real‐time RT‐PCR (TaqMan®).

Results
There was a clear increase in the treated AR patients, in comparison with untreated AR patients, in the expression of both IL‐10 (mean change from baseline (SEM): 3.1 (0.8) vs. −0.3 (0.3), P<0.002, Mann–Whitney U‐test) and IL‐18 (2.7 (0.9) vs. −0.2 (0.6), P<0.03) mRNA after 1 year. The clearest increase in IL‐10 mRNA expression was seen in patients who did not benefit at all (6.0 (2.3), P<0.001 vs. untreated) and the least increase in patients that had the greatest reduction of symptoms (0.8 (0.6), n.s. vs. untreated) at 1 year. The clearest increase in IL‐18 mRNA expression was seen in patients with moderate outcome (3.4 (1.6), P<0.04 vs. untreated). In intermediate samples, taken when the maintenance dose was reached, the peak expression of allergen‐induced IL‐10 mRNA was associated with the most favourable outcome of SIT (P=0.01, Fisher exact test). A similar trend was seen in IL‐18 mRNA expression.

Conclusions
The results suggest that an early and transient increase in allergen‐specific IL‐10 and IL‐18 mRNA expression in PBMC is essential for the therapeutic outcome after 1 year of SIT.
Glucocorticoids up‐regulate constitutive interleukin‐10 production by human monocytes
Authors: Mozo L. , Suárez A. , Gutiérrez C.
Published: March 2004

Background
IL‐10 plays an immunosuppressive role in inflammatory responses. Increased plasma levels of IL‐10 have been detected in patients under glucocorticoid (GC) therapy, indicating that steroids may exert their suppressive effect, in part, by increasing IL‐10 production.

Objectives
The aim was to define possible mechanisms by which steroids up‐regulate IL‐10 production. To this end, we have analysed ex vivo the effect of GCs on the constitutive production of IL‐10 by lymphocytes and cells of myeloid origin.

Methods
Monocytes and T cells were isolated by a Percoll gradient and B cells were purified by rosetting. Protein and mRNA IL‐10 levels were determined by ELISA and by Northern blot, respectively.

Results
Monocytes, but not T or B cells, up‐regulated the constitutive production of IL‐10 following pre‐treatment for at least 12 h with physiological doses of dexamethasone (Dex). Up‐regulation of IL‐10 occurred at both protein and mRNA levels, probably indicating that the effect of Dex was by incrementing gene transcription. Other steroids had similar outcomes, their effects being dose‐related, proportional to the steroid potency and totally reversed by the steroid antagonist RU486. Thus, transcript levels of IL‐10 were up‐regulated by GCs probably through binding of the GC receptor to its specific glucocorticoid response element sequence in the IL‐10 promoter. In contrast to monocytes, differentiated immature macrophages and dendritic cells did not vary their constitutive IL‐10 production after pre‐treatment with Dex.

Conclusion
Our results support the fact that steroids up‐regulate constitutive IL‐10 production by selectively triggering activation signals on monocytes.
Association between polymorphisms in serine protease inhibitor, kazal type 5 and asthma phenotypes in a large German population sample
Authors: Kabesch M. , Carr D. , Weiland S. K. , Von Mutius E.
Published: March 2004

Background
Atopic diseases are characterized by immunoglobulin E (IgE)‐mediated immune responses towards common allergens, many of which are proteases. Recently it has been suggested that a proteinase inhibitor gene, SPINK5, which is located on chromosome 5q31, may play a role in the pathogenesis of atopic diseases.

Objective
We investigated the association between the polymorphism G1258A leading to a putative amino acid change (Glu420Lys) in serine protease inhibitor, kazal type 5 (SPINK5) and phenotypes of atopic diseases in a large general population sample of German children.

Methods
Parental questionnaires were used and children underwent skin prick testing, pulmonary function testing and bronchial challenge. Blood was collected for serum IgE measurements and DNA extraction. In total, 1161 children were genotyped for the SPINK5 Glu420Lys polymorphism and association studies were performed.

Results
A significant association between SPINK5 420Lys and the development of asthma was observed (OR 1.77; 95%CI: 1.02–3.06, P=0.041 for 420Lys homocygotes). Atopic carriers of SPINK5 420Lys showed an increased risk for asthma and asthma symptoms (OR 2.06; 95%CI: 1.01–4.20, P=0.047). When children with a combination of asthma and atopic dermatitis were compared with normal controls, the SPINK5 420Lys genotype was more prevalent in the disease group (OR 4.56; 95%CI: 1.370–15.12, P=0.007). No association between SPINK5 420Lys genotypes and total serum IgE levels, skin prick test (SPT) reactivity or atopic dermatitis was observed.

Conclusion
These results suggest that SPINK5 Glu420Lys polymorphism may be associated with certain asthma phenotypes characterized by the concomitant expression of asthma and atopic dermatitis or SPT reactivity.
Bacillus Calmette–Guérin‐induced interleukin‐12 did not additionally improve clinical and immunologic parameters in asthmatic children treated with sublingual immunotherapy
Authors: Arikan C. , Bahceciler N. N. , Deniz G. , Akdis M. , Akkoc T. , Akdis C. A. , Barlan I. B.
Published: March 2004

Objective
To evaluate the effect of bacillus Calmette–Guérin (BCG) as an adjuvant to specific sublingual immunotherapy (SLIT) on the cytokine profile of peripheral blood mononuclear cells (PBMCs) and clinical outcome.

Methods
Thirty‐two children with asthma and rhinitis allergic to house dust mite (HDM) with negative purified protein derivative (PPD) skin test response were enrolled. After a run‐in period of 8 weeks, patients were randomized to receive either SLIT only (n=16) or one dose of BCG immunization before initiation of SLIT (n=16) with a standardized Dermatophagoides pteronyssinus (D. pteronyssinus)+D. farinea 50/50 extract. PPD‐negative asthmatics (n=5) allergic to HDM receiving inhaled therapy only were included for comparison of cytokine levels in PBMC cultures. Efficacy was assessed both at the end of run‐in and 6 months of treatment periods with criteria including symptom, medication and quality‐of‐life (QoL) scores, IgE levels, lung function, provocation concentration (PC20), eosinophil count and skin prick tests. IL‐4, IL‐5, IL‐10, IL‐12, IL‐13 and IFN‐γ levels were determined in antigen specifically and polyclonally stimulated PBMC cultures.

Results
Both treatment groups showed significant improvement at the end of 6 months for asthma and rhinitis scores and QoL, number of asthma attacks, amount of β2‐agonists, inhaled and intranasal steroids, blood eosinophil counts and PC20. Interestingly, phytohaemagglutinin (PHA)‐stimulated IL‐12 and D. pteronyssinus‐stimulated IFN‐γ in PBMC were significantly higher in the treatment groups than controls. In addition, IL‐12 levels in response to D. pteronyssinus and PHA stimulation were significantly higher in the SLIT+BCG group than the SLIT alone group and controls.

Conclusion
The present study demonstrates that successful SLIT is parallel to increased IFN‐γ production by PBMC. Although simultaneous BCG vaccination enhanced IL‐12 production, it did not additionally improve the clinical outcome.
Determinants of endotoxin levels in living environments of farmers' children and their peers from rural areas
Authors: Waser M. , Schierl R. , Von mutius E. , Maisch S. , Carr D. , Riedler J. , Eder W. , Schreuer M. , Nowak D. , Braun‐fahrländer C. ,   the ALEX Study Team
Published: March 2004

Background
Lower frequencies of asthma and hayfever have been observed in children with contact to livestock. At school age, the amount of endotoxin measured in the dust of children's mattresses is inversely related to the occurrence of atopic asthma, hayfever and atopic sensitization both in children from farming and non‐farming households.

Objective
The aim of the present study was to investigate which home and lifestyle characteristics of farm and non‐farm families contribute to endotoxin levels measured in different indoor home environments.

Methods
In the framework of the Allergy and Endotoxin (ALEX) Study, endotoxin was measured in dust samples from the living room floor and the child's mattress of 319 farmers' families and 493 non‐farming families, and in settled dust from stables. Endotoxin content of all dust samples was determined by a kinetic Limulus assay (Limulus‐Amebocyte‐Lysate test). Information about the child's activities on farms, home characteristics and cleaning behaviours was obtained from parental questionnaires.

Results
Endotoxin levels in stables did not predict the amount of endotoxin measured in floors or mattresses. However, a dose‐dependent association between the child's activity on the farm and indoor home endotoxin levels was observed, both in farm and non‐farm children. In non‐farm children pet keeping and the frequency of floor cleaning were additionally associated with endotoxin levels, whereas in farm children parental farm activities, study area, time since last cleaning, the mattress type as well as younger age of the children contributed to increased microbial exposure.

Conclusion
These results demonstrate that regular contact to farm animals increases indoor home endotoxin concentrations, both in farm and non‐farm children, and might thus explain the protective effect of contact to livestock on atopic outcomes. To assess children's individual exposure to a microbial environment, measures of mattress dust exposure are needed as stable endotoxin concentrations were not associated with indoor home levels.
Pruritus‐associated response mediated by cutaneous histamine H3 receptors
Authors: Sugimoto Y. , Iba Y. , Nakamura Y. , Kayasuga R. , Kamei C.
Published: March 2004

Background  Histamine is one of the most common chemical mediators causing pruritus, and H1 receptor antagonists have been used as a first choice in its treatment. On the other hand, although the presence of H3 receptors has been identified in the skin, few studies have investigated the involvement of H3 receptors on pruritus.

Objective  The purpose of this study was to examine whether H3 receptor agonist or antagonist influences the incidence of scratching behaviour in ICR or mast cell‐deficient WBB6F1‐W/WV mice.

Methods  The mice were given an intradermal injection of H3 receptor agonist or antagonist into the rostral part of the back, and the occurrence of scratching behaviour at the injected site by the hind paws was counted over 60 min.

Results  H3 receptor antagonists, thioperamide and AQ0145 significantly increased the incidence of scratching behaviour in ICR mice. H3 receptor agonist, (R)‐alpha‐methylhistamine, had no effect. On the other hand, (R)‐alpha‐methylhistamine significantly inhibited thioperamide or AQ0145‐induced scratching behaviour. In addition, both thioperamide and AQ0145 elicited scratching behaviour in mast cell‐deficient WBB6F1‐W/WV mice.

Conclusion  From these results, it may be concluded that H3 receptors are involved in the modulation of pruritus in the skin, and mast cells are not essential in this response. In addition, H3 receptor agonists can be useful as a novel therapeutic approach against pruritus.
Protein sequence analysis of a novel 103‐kDa Dermatophagoides pteronyssinus mite allergen and prevalence of serum immunoglobulin E reactivity to rDer p 11 in allergic adult patients
Authors: Lee C.‐S. , Tsai L.‐C. , Chao P.‐L. , Lin C.‐Y. , Hung M.‐W. , Chien A.‐I. , Chiang Y.‐T. , Han S.‐H.
Published: March 2004

Background
House dust mites are regarded as important indoor allergens. While the most studies mite allergens are low molecular weight (mw), a high mw Dermatophagoides farinae mite paramyosin (Der f 11) has recently been cloned. We have also cloned a novel high mw Dermatophagoides pteronyssinus (Dp) mite allergen, Der p 11.

Objective
The aim of this study was to isolate and express a cDNA gene coding for a Der p 11 allergen, to compare the sequence of Der p 11 with other antigens and to evaluate the presence of IgE reactivity to the recombinant protein (rDer p 11) in the sera of allergic adult patients.

Methods
The full‐length Der p 11 gene was isolated by cDNA library screening, 5′‐3′ rapid amplification of cDNA ends and PCR. The cDNA gene was expressed as a glutathione‐S‐transferase fusion protein in Escherichia coli. The allergenicity of rDer p 11 was tested by human IgE immunodot or immunoblot assay in a large panel of 100 allergic patients with bronchial asthma, allergic rhinitis or eczema.

Results
Der p 11 is a 2965 bp cDNA gene with a 2625 bp open reading frame coding for a 875 amino acid protein. The deduced amino acid sequence of the Der p 11 showed significant homology with various invertebrate paramyosins. The prevalence of serum IgE reactivity to rDer p 11 on immunodot assay ranged from 41.7% to 66.7% in different allergic patient groups, whereas it was rare in non‐atopic patients with urticaria (18.8%) and in normal individuals (8%). A high frequency (five out of eight) of MAST(Dp)− allergic serum samples had specific IgE‐binding activity to rDer p 11 or its fragments on immunoblot assay, even though their IgE‐binding activity to Dp extract was either weak or negative.

Conclusion
The 103‐kDa Der p 11 appears to be major Dp mite allergen with a high frequency of IgE reactivity in sera of patients allergic to mites.
Bronchoalveolar lavage fluid concentrations of transforming growth factor (TGF)‐β1, TGF‐β2, interleukin (IL)‐4 and IL‐13 after segmental allergen challenge and their effects on α‐smooth muscle actin and collagen III synthesis by primary human lung fibroblasts
Authors: Batra V. , Musani A. I. , Hastie A. T. , Khurana S. , Carpenter K. A. , Zangrilli J. G. , Peters S. P.
Published: March 2004

Rationale
Asthmatic airway remodelling is characterized by myofibroblast hyperplasia and subbasement membrane collagen deposition. We hypothesized that cytokines and growth factors implicated in asthmatic airway remodelling are increased in bronchoalveolar lavage (BAL) fluid of asthmatics after segmental allergen challenge (SAC), and that these growth factors and cytokines increase α‐smooth muscle actin (α‐SMA) and collagen III synthesis by human lung fibroblasts (HLFs).

Methods
Transforming growth factor (TGF)‐β1, TGF‐β2, IL‐4 and IL‐13 levels were measured in BAL fluid from 10 asthmatics and 9 non‐asthmatic controls at baseline and then 1 day, 1 week and 2 weeks after SAC. Confluent cultures of HLFs were stimulated by exogenous addition of TGF‐β1, TGF‐β2, IL‐4 or IL‐13 (concentration range 0.01–10 ng/mL) over 48 h. Collagen III was measured in culture supernates and α‐SMA in cell lysates by Western blot.

Results
At baseline, there was no difference in BAL fluid concentrations of TGF‐β1, IL‐4 and IL‐13 between asthmatics and controls; however, non‐asthmatics had higher concentrations of total TGF‐β2. In asthmatics, BAL fluid concentrations of all four factors increased significantly 1 day after SAC. TGF‐β1, TGF‐β2 and IL‐13 concentrations returned to baseline by 1 week after SAC, but BAL fluid IL‐4 concentration remained elevated for at least 2 weeks. TGF‐β1, TGF‐β2 and IL‐4 significantly increased α‐SMA in fibroblasts, but only IL‐4 caused corresponding increases in collagen III synthesis. IL‐13 had no direct effects on collagen III synthesis and α‐SMA expression.

Conclusions
Because IL‐4 caused a dose‐dependent increase in α‐SMA and collagen III synthesis, it may be an important cytokine mediating asthmatic airway remodelling. TGF‐β1 and TGF‐β2 may also play a role in airway remodelling by stimulating phenotypic change of fibroblasts to myofibroblasts. Additionally, collagen III synthesis appears to be independent of myofibroblast phenotype and is apparently regulated by different growth factors and cytokines.
Increased airway responsiveness, allergy‐type‐I skin responses and systemic anaphylaxis in a humanized‐severe combined immuno‐deficiency mouse model
Authors: Herz U. , Botchkarev V. A. , Paus R. , Renz H.
Published: March 2004

Background
In patients with allergic bronchial asthma, a strong relationship between elevated serum IgE antibody titres and the development of increased airway responsiveness (AR) has been demonstrated. To further elucidate the relationship between human (hu) IgE and development of increased AR, we developed an in vivo model utilizing immuno‐compromised severe combined immuno‐deficiency (SCID) mice.

Methods
SCID mice were either reconstituted with peripheral blood mononuclear cells (PBMC) from non‐atopic, healthy or atopic individuals sensitized against house dust mite allergen (Der p), or passively sensitized with plasma from non‐atopic, healthy or atopic individuals.

Results
In both systems, atopic hu‐SCID mice developed increased AR. The following results suggest that these responses were mediated via IgE antibodies: increased AR did not occur after transfer of either PBMC or IgE‐negative plasma from non‐atopic individuals; increased AR occurred simultaneous with increased serotonin release detected 15 min after allergen–aerosol challenge in bronchoalveolar lavage fluid; and increased AR required at least two allergen–aerosol challenges. SCID mice reconstituted with serum containing anti‐Der p IgE antibodies developed positive immediate‐type skin test responses to intradermal injection of Der p as well as anti‐hu‐IgE antibody. In addition, IgE binding to skin mast cells was demonstrated by immunohistochemistry. Furthermore, intravenous challenge of hu anti‐Der p positive SCID mice with Der p resulted in systemic anaphylaxis.

Conclusion
These data provide evidence that passive immunization of SCID mice with hu IgE alters AR and that T cells and eosinophils were not a requirement for the development of increased AR in this model.
Association of polymorphisms in Th1, Th2 cytokine genes with hayfever and atopy in a subsample of EPIC‐Heidelberg
Authors: Nieters A. , Linseisen J. , Becker N.
Published: March 2004

Background
Hayfever is determined by an interaction of environmental and genetic factors and biologically characterized by an imbalanced T helper cell 1 (Th1) and Th2 immune response and elevated IgE levels against inhalant allergens. Indications exist that polymorphisms in cytokine genes involved in the regulation of the Th1/Th2 balance may contribute to the allergic phenotype.

Objective
We investigate whether polymorphisms in genes directly or indirectly involved in Th1, Th2 immune response are associated with hayfever and elevated IgE levels against inhalant allergens.

Methods
From a subsample of the European Prospective Investigation into Cancer and Nutrition‐Heidelberg, 322 subjects with hayfever and 322 age‐ and sex‐matched non‐allergic controls were selected and genotypes determined for 15 polymorphisms in 13 genes. Plasma IgE against inhalant allergens was measured via CAPSX1 (Phadiatop) test. We computed odds ratios (ORs) by logistic regression, tests on group differences of IgE‐levels in dependence upon genotype and tests for trend by means of non‐parametric methods.

Results
We found decreased OR for hayfever (OR=0.60, 95%CI=0.4–0.9) and sensitization to inhalant antigens (OR=0.5, 95%CI=0.4–0.8) in heterozygotes of the IL‐6 −174 G/C polymorphism. Homozygotes of the G‐allele in IL‐2 −330 T/G were at increased risk of hayfever (OR=2.6, 95%CI=1.3–5.2). Moreover, we found indications for differences in IgE levels against inhalant allergens in dependence upon genotypes of polymorphisms in IL‐4R, IL‐6, IL‐13, and IL‐18.

Conclusion
Our data suggest an association of genetic variants in IL‐6 and IL‐2 with hayfever, confirm a role of polymorphisms in IL‐4R, IL‐13, and IL‐18 for the elevated IgE phenotype, and add IL‐6 to the list of candidate genes.
The relation of markers of fetal growth with asthma, allergies and serum immunoglobulin E levels in children at age 5–7 years
Authors: Bolte G. , Schmidt M. , Maziak W. , Keil U. , Nasca P. , Von Mutius E. , Weiland S. K.
Published: March 2004

Background
It has been suggested that fetal growth and maturation have an impact on the development of allergic diseases later in life.

Objective
To examine the association between measures of fetal growth and allergic disease in children at age 5–7 years.

Methods
As part of the German International Study of Asthma and Allergies in Childhood phase II surveys, a random sample of school beginners (n=1138) was examined in 1995. Data on anthropometric measures at birth and gestational age were obtained from maternal copies of birth records. Data on symptoms and doctor‐diagnosed asthma, atopic dermatitis and hayfever were gathered by parental questionnaires. Atopic sensitization was assessed by serum IgE and skin prick tests to common aeroallergens. Children (741) had complete data for the explanatory variables of interest and were thus eligible for this analysis. Confounder‐adjusted prevalence odds ratios (PORs) and means ratios with 95% confidence intervals (CI) were calculated using multiple logistic and linear regression.

Results
Birth weight and gestational age were positively associated with atopic sensitization (Ptrend=0.025 and 0.035, respectively). Children with a low birth weight relative to head circumference had a decreased risk of sensitization (POR 0.44, 95% CI 0.21–0.91; Ptrend=0.020). Moreover, total serum IgE increased with increasing birth weight (Ptrend=0.042). No consistent relationship was observed between markers of fetal growth and wheezing, doctor‐diagnosed asthma, atopic dermatitis and hayfever.

Conclusion
These data suggest that fetal growth and maturity are associated with atopic sensitization and total serum IgE levels in childhood.
In vitro allergy diagnosis: should we follow the flow?
Authors: Ebo D. G. , Hagendorens M. M. , Bridts C. H. , Schuerwegh A. J. , De clerck L. S. , Stevens W. J.
Published: March 2004

During the last 5 years, an increasing number of studies have demonstrated that flow cytometric quantification of in vitro basophil activation can be a quite performant and reliable tool to measure IgE‐dependent allergen‐specific responses in allergic patients. So far, most assays have used CD63 as a basophil activation marker and native allergen extracts for stimulation. However, other basophil markers and recombinant allergens have recently been introduced. The technique has been applied for the diagnosis of allergy to pollen, house dust mite, food, natural rubber latex, hymenoptera venom and drugs. In addition, the technique has proven to be useful in non‐IgE‐mediated reactions such as hypersensitivity to drugs as well as detection of auto‐antibodies in chronic urticaria. This review will focus on some specific issues: (1) principles of flow cytometric analysis of in vitro‐activated basophils, (2) general technical aspects of the technique (including passive sensitization), (3) clinical applications and (4) recommendations for further development and evaluation of the technique.
The recombinant major allergen of Parietaria judaica and its hypoallergenic variant: in vivo evaluation in a murine model of allergic sensitization
Authors: Orlandi A. , Grasso F. , Corinti S. , Marinaro M. , Bonura A. , Boirivant M. , Colombo P. , Di Felice G.
Published: March 2004

Background  Par j 1 represents the major allergenic component of Parietaria judaica pollen. Its three‐dimensional structure is stabilized by four disulphide bridges. A family of three‐dimensional mutants of the recombinant Par j 1 (rPar j 1) allergen, showing reduced allergenicity and retained T cell recognition has been recently developed by site‐directed mutagenesis.

Objective  To develop and characterize a murine model of IgE sensitization to rPar j 1. To evaluate similarities between the murine model and the human IgE response. To investigate in this model the recognition of a hypoallergenic mutant of Par j 1, and to study the immune responses elicited in mice by the mutant itself.

Methods  BALB/c mice were sensitized by two intraperitoneal immunizations with rPar j 1 in alum on days 0 and 21. Allergen‐specific serum IgE and IgG responses were studied by direct ELISA and immunoblotting, ELISA inhibition and competitive ELISA. Cell proliferation was evaluated in splenocyte cultures.

Results  Sensitization with rPar j 1 induced high levels of IgE and IgG1 vs. low levels of IgG2a. Mouse antibodies specific to rPar j 1 were able to compete with human IgE for recognition of rPar j 1. IgE from mice immunized with rPar j 1 showed a significantly reduced binding activity towards the hypoallergenic variant rPjC, which lacks three disulphide bridges. On the contrary, rPjC was recognized by IgG1 and IgG2a antibodies as well as rPar j 1. The proliferative response to rPjC by splenocytes from mice immunized with rPar j 1 was comparable to that stimulated by rPar j 1. Immunization with rPjC induced low levels of IgE antibodies to the rPjC itself, while IgG and proliferative responses were similar to those induced by rPar j 1.

Conclusion  Conformational variants of allergens, displaying reduced allergenicity accompanied by retained IgG and T cell recognition, offer a safe, specific and flexible approach to immunotherapy of type I allergy. Our mouse model of IgE sensitization to a recombinant allergen, mimicking the human response to its native counterpart, could provide valuable information for pre‐clinical testing of such hypoallergenic molecules.
Prolonged allergen challenge in mice leads to persistent airway remodelling
Authors: Mcmillan S. J. , Lloyd C. M.
Published: March 2004

Background
Inflammatory infiltrates, airway hyper‐responsiveness, goblet cell hyperplasia and subepithelial thickening are characteristic of chronic asthma. Current animal models of allergen‐induced airway inflammation generally concentrate on the acute inflammation following allergen exposure and fail to mimic all of these features.

Objective
The aim of this study was to use a murine model of prolonged allergen‐induced airway inflammation in order to characterize the cells and molecules involved in the ensuing airway remodelling. Moreover, we investigated whether remodelling persists in the absence of continued allergen challenge.

Methods
Acute pulmonary eosinophilia and airways hyper‐reactivity were induced after six serial allergen challenges in sensitized mice (acute phase). Mice were subsequently challenged three times a week with ovalbumin (OVA) (chronic phase) up to day 55. To investigate the persistence of pathology, one group of mice were left for another 4 weeks without further allergen challenge (day 80).

Results
The extended OVA challenge protocol caused significant airway remodelling, which was absent in the acute phase. Specifically, remodelling was characterized by deposition of collagen as well as airway smooth muscle and goblet cell hyperplasia. Importantly, these airway changes, together with tissue eosinophilia were sustained in the absence of further allergen challenge. Examination of cytokines revealed a dramatic up‐regulation of IL‐4 and tumour growth factor‐β1 during the chronic phase. Interestingly, while IL‐4 levels were significantly increased during the chronic phase, levels of IL‐13 fell. Levels of the Th1‐associated cytokine IFN‐γ also increased during the chronic phase.

Conclusion
In conclusion, we have demonstrated that prolonged allergen challenge results in persistent airway wall remodelling.
Low numbers of interleukin‐12‐producing cord blood mononuclear cells and immunoglobulin E sensitization in early childhood
Authors: Nilsson C. , Larsson A.‐K. , Höglind A. , Gabrielsson S. , Blomberg M. Troye , Lilja G.
Published: March 2004

Background
Successful pregnancies are associated with skewing towards a Th2 cytokine profile. Cytokine responses to allergens can be detected in cord blood mononuclear cells (CBMC), suggesting allergen priming already in utero.

Objective
To investigate the cytokine profile in CBMC after in vitro stimulation with allergens and to relate the responses to the outcome in terms of allergic disease at 2 years of age.

Methods
CBMC were isolated from 82 children. The responses to ovalbumin (OVA), birch, cat and phytohaemagglutinin (PHA) were investigated by the ELISpot technique. The numbers of IFN‐γ‐, IL‐4‐ and IL‐12‐producing CBMC were counted for each stimulation. The children were followed prospectively; skin prick test (SPT) and RAST towards food and inhalant allergens were assessed at 24 months of age.

Results
Sixteen (19.5%) children were classified as IgE sensitized (positive SPT; 3 mm and/or RAST; 0.35 kUA/L). The numbers of IL‐12‐producing CBMC after stimulation with birch, OVA and cat were lower among IgE‐sensitized children, statistically significant for cat. IFN‐γ‐producing cells, did not differ in numbers between the sensitized and non‐sensitized children. Children who had atopic eczema/dermatitis syndrome (AEDS) during the observation (n=53) had significantly lower numbers of IFN‐γ‐producing CBMC after stimulation with OVA and cat than their non‐AEDS counterparts.

Conclusions
Although the numbers of infants in our study are limited our data suggest that a low number of IL‐12‐producing CBMC is associated with IgE sensitization during early childhood and that a reduced number of IFN‐γ‐producing CBMC promotes the development of AEDS during the first 2 years of life.
Identification by immunoblot of venom glycoproteins displaying immunoglobulin E‐binding N‐glycans as cross‐reactive allergens in honeybee and yellow jacket venom
Authors: Hemmer W. , Focke M. , Kolarich D. , Dalik I. , Götz M. , Jarisch R.
Published: March 2004

Background
IgE antibodies against carbohydrate epitopes have been identified recently as a major cause of in vitro double positivity to honeybee (HB) and vespid venom in patients with stinging‐insect allergy. As these antibodies possibly have low clinical relevance they may be misleading in the diagnosis of venom allergy.

Objective
To confirm the role of carbohydrate epitopes in double positivity and to locate the responsible glycoallergens in HB and yellow jacket (YJ) venom by western blot.

Methods
Immunoblot inhibition using HB venom, YJ venom and two glycoprotein sources displaying 1‐3‐fucosylated N‐glycans (i.e. oilseed rape (OSR) pollen, and the synthetic neo‐glycoprotein fucosylated/xylosylated N‐glycans from bromelain coupled to bovine serum albumin (MUXF‐BSA)) as inhibitors were performed with sera from 15 double‐positive patients with stinging‐insect allergy. Additionally, reactivity with blotted hymenoptera venoms of a carbohydrate‐specific rabbit antiserum against OSR pollen was investigated.

Results
Major venom glycoallergens binding with carbohydrate‐specific human IgE and rabbit IgG were detected in HB venom at 42 (hyaluronidase (HYA)), 46, 65 and 95 kDa, and in YJ venom at 38 and 43 kDa (HYA). Antibody binding to these allergens was completely lost after periodate treatment. Glycans of HB phospholipase were bound by patients' IgE only after protein denaturation. In 10 of the 15 patients the reactivity was with the second venom because of carbohydrates alone. The high‐molecular‐weight glycoallergens identified in HB venom probably correspond to similar proteins described earlier, including allergens B and C. The 38‐kDa YJ allergen might represent a homologue of V mac 3.

Conclusions
The data confirm the proposed role of carbohydrate‐specific IgE in double positivity to HB and YJ venom and shed new light on some previously described minor hymenoptera allergens of uncertain clinical significance. The consideration of carbohydrate‐specific IgE may allow to discriminate between patients with potentially relevant and patients with non‐relevant double sensitization.
Investigation into the mechanisms by which nedocromil sodium, frusemide and bumetanide inhibit the histamine‐induced itch and flare response in human skin in vivo
Authors: Willis E. F. , Clough G. F. , Church M. K.
Published: March 2004

Background
In a previous study, iontophoresis of nedocromil sodium into human skin in vivo was shown to reduce histamine‐induced itch and flare. In asthma, the Na+/K+/2Cl− cotransporter inhibitors, frusemide and bumetanide, have been reported to have many similar actions to nedocromil sodium.

Objective
To compare the effects of these drugs in the histamine‐induced itch, flare and weal response in human skin in vivo and elucidate their site of action.

Methods
Nedocromil sodium, frusemide bumetanide and reversed osmosis water (control), were introduced by iontophoresis into the forearm skin of 10 volunteers in each of two single‐blind studies. In study 1, histamine (20 μL of 100 μm) or vehicle was injected into the area of iontophoresis 10 min later. In study 2, histamine or vehicle was injected 5 mm outside the area of iontophoresis so the flare developed over the area of iontophoresis. Itch was scored on a visual analogue scale every 20 s for 5 min, flare areas were assessed using scanning laser Doppler imaging up to 10 min and weal was assessed by planimetry at 10 min.

Results
In study 1, nedocromil sodium, frusemide and bumetanide reduced itch scores by 36%, 48% and 34%, respectively, and flare areas by 17%, 26% and 15% respectively (all P<0.05). Weal areas and blood flux in the flare were unaffected. In study 2, itch scores, flare areas and weal areas were not inhibited. Also, blood flux values in areas of drug and water iontophoresis were not different.

Conclusion
This study has provided evidence to support the hypothesis that nedocromil sodium, frusemide and bumetanide inhibit sensory nerve activation to reduce the itch and flare responses induced by histamine in human skin in vivo. It is likely that inhibition of a Na+/K+/2Cl− cotransporter in the sensory nerve membrane is a possible mechanism of action.
A double‐blind, placebo‐controlled birch allergy vaccination study: inhibition of CD23‐mediated serum‐immunoglobulin E‐facilitated allergen presentation
Authors: Van Neerven R. J. J. , Arvidsson M. , Ipsen H. , Sparholt S. H. , Rak S. , Würtzen P.A.
Published: March 2004

Background

The clinical efficacy of specific allergy vaccination (SAV), previously called specific immunotherapy is well documented. The working mechanism of this treatment is not completely known at present. Allergen‐specific CD4+ T lymphocytes are activated at extremely low allergen concentrations in vivo possibly as a result of serum IgE‐facilitated allergen presentation (S‐FAP). Previously, we have shown that this process can be inhibited by long‐term birch SAV sera.

Methods

In the present study, we have analysed sera from birch‐allergic patients in a randomized double‐blind, placebo‐controlled clinical trial for their ability to mediate S‐FAP. Birch‐specific IgE levels were not changed after SAV. Bet v 1‐specific IgE levels, however, were significantly decreased (P<0.05) and Bet v 1‐specific IgG4 levels were strongly increased after SAV (P<0.001). None of these changes were observed in the placebo group. When the sera were tested for their ability to induce S‐FAP, a complete abrogation of this effect was noted in the sera from patients receiving active treatment (P<0.001), but not in the control group. This inhibition of S‐FAP seemed to be associated with the reduction in the ratio between Bet v 1‐specific IgE and IgG4 antibodies in serum, but a clear correlation could not be demonstrated.

Conclusion

In conclusion, the present study clearly shows that SAV leads to an inhibition of the S‐FAP needed to obtain optimal T cell activation at the low allergen concentrations present in vivo. This novel mechanism may explain the increased allergen threshold levels found in allergen provocation tests and the reduction of late‐phase reactions observed after SAV.
VH gene usage in immunoglobulin E responses of seasonal rhinitis patients allergic to grass pollen is oligoclonal and antigen driven
Authors: Davies J. M. , O'Hehir R. E.
Published: March 2004

Background
IgE is the pivotal‐specific effector molecule of allergic reactions yet it remains unclear whether the elevated production of IgE in atopic individuals is due to superantigen activation of B cell populations, increased antibody class switching to IgE or oligoclonal allergen‐driven IgE responses.

Objectives
To increase our understanding of the mechanisms driving IgE responses in allergic disease we examined immunoglobulin variable regions of IgE heavy chain transcripts from three patients with seasonal rhinitis due to grass pollen allergy.

Methods
Variable domain of heavy chain‐epsilon constant domain 1 cDNAs were amplified from peripheral blood using a two‐step semi‐nested PCR, cloned and sequenced.

Results
The VH gene family usage in subject A was broadly based, but there were two clusters of sequences using genes VH 3‐9 and 3‐11 with unusually low levels of somatic mutations, 0–3%. Subject B repeatedly used VH 1‐69 and subject C repeatedly used VH 1‐02, 1‐46 and 5a genes. Most clones were highly mutated being only 86–95% homologous to their germline VH gene counterparts and somatic mutations were more abundant at the complementarity determining rather than framework regions. Multiple sequence alignment revealed both repeated use of particular VH genes as well as clonal relatedness among clusters of IgE transcripts.

Conclusion
In contrast to previous studies we observed no preferred VH gene common to IgE transcripts of the three subjects allergic to grass pollen. Moreover, most of the VH gene characteristics of the IgE transcripts were consistent with oligoclonal antigen‐driven IgE responses.
RNA editing of interleukin‐12 receptor β2, 2451 C‐to‐U (Ala 604 Val) conversion, associated with atopy
Authors: Kondo N. , Matsui E. , Kaneko H. , Aoki M. , Kato Z. , Fukao T. , Kasahara K. , Morimoto N.
Published: March 2004

Background
The production of IgE in B lymphocytes is down‐regulated by IFN‐γ. IL‐12 induces IFN‐γ production by T lymphocytes and natural killer cells by binding to its specific receptor. RNA editing is a post‐transcriptional modification.

Objective
Here we show that the RNA editing of IL‐12 receptor (R) β2 is associated with atopy.

Methods
Atopic patients and non‐atopic healthy controls were studied. Fragments of IL‐12R β2 cDNA and genomic DNA were amplified and sequenced. Furthermore, the function of the IL‐12R β2 chain was investigated.

Results
Sequence analysis of the cDNA clones representing IL‐12R β2 mRNA transcripts revealed a C‐to‐U conversion at nucleotide 2451 (Ala 604 Val) on exon 13 in some atopic patients. Surprisingly, sequence analysis of their genomic DNA showed no 2451 C‐to‐T (Ala 604 Val) mutation. We concluded that the observed C‐to‐U mismatch in the cDNA clone is due to a post‐transcriptional modification, RNA editing. The C‐to‐U conversion was observed in 21 (20.6%) of 102 atopic patients, whereas this conversion was observed in only 4 (3.8%) of 104 non‐atopic subjects (P<0.001). IFN‐γ production by peripheral blood mononuclear cells (PBMCs) stimulated with IL‐12 in the subjects with the C‐to‐U conversion was significantly lower than that in the subjects without the C‐to‐U conversion. In atopic patients with the C‐to‐U conversion, PBMCs faintly showed the tyrosine phosphorylation of Stat4, and the IgE production by PBMCs was not suppressed by IL‐12 whereas it was suppressed by IFN‐γ.

Conclusions
The RNA editing of IL‐12R β2, 2451 C‐to‐U (Ala 604 Val) conversion causes impairment of the IL‐12 signal cascade and the subsequent reduction in IFN‐γ production, resulting in the impaired down‐regulation of IgE production. This is the first report indicating that atopy is associated with RNA editing.
Interleukin‐4 increases the permeability of human endothelial cells in culture
Authors: Kotowicz K. , Callard R. E. , Klein N. J. , Jacobs M. G.
Published: March 2004

Background
IL‐4 plays a key role in the induction of allergic inflammation, but its role as an effector molecule is less well‐established. Although some observations suggest that IL‐4 may mediate increased vascular permeability, which is a characteristic feature of allergic inflammation, evidence for a direct effect on endothelial cell permeability is lacking.

Objective
To determine the effect of human IL‐4 on the albumin permeability of cultured human endothelial cells.

Methods
Human umbilical vein endothelial cells were cultured on permeable membranes and the albumin permeability of endothelial monolayers was measured with and without exposure to recombinant human IL‐4. Endothelial cells were exposed to various concentrations of IL‐4 (0.001–100 U/mL), for various durations (6–24 h), either in the presence or absence of anti‐IL‐4 antibody. Recovery of endothelial barrier function following exposure to IL‐4 was also examined.

Results
IL‐4 induced a dose‐dependent, reversible increase in endothelial permeability to albumin. Low concentrations of IL‐4 (1 U/mL) induced a significant increase in endothelial permeability (P=0.004). IL‐4‐mediated endothelial leak occurred rapidly, within 6 h of exposure.

Conclusions
IL‐4 has the capacity to induce vascular leak by a direct effect on cultured endothelial cells, suggesting a potential effector role for IL‐4 in the pathogenesis of vascular leak in allergic diseases.
New pregnancies and loss of allergy
Authors: Harris J. M. , White C. , Moffat S. , Mills P. , Newman Taylor A. J. , Cullinan P.
Published: March 2004

Background
Inverse associations between allergic disease and sibship have been consistently described and are frequently explained by purported lower rates of early infection among children from small families. Alternative explanations include the possibility that pregnancy itself determines maternal atopic status.

Objective
To test the hypothesis that atopy defined by skin prick test (SPT) declines with increasing numbers of pregnancies.

Methods
At enrolment to a birth cohort, mothers were skin prick tested to three common allergens. Seven years later these women underwent a second SPT and provided information on their reproductive histories. At both visits, information on allergic disease was also sought.

Results
Twenty five (15%) women who were initially atopic were no longer so at the second visit; loss of hayfever symptoms was reported by 33 (29%) women. Women with higher numbers of intervening pregnancies were more likely to ‘lose’ their atopy (P=0.05) and symptoms of hayfever (P=0.02); this was not true for asthma. The findings could not be accounted for by maternal age.

Conclusion
Successive pregnancies may in part determine a mother's atopic state. Since maternal atopy is a risk factor for childhood atopic disease, this process may affect the atopic state of successive children. These findings suggest an alternative explanation for the sibship effect in allergic disease.
Respiratory syncytial virus, pneumonia virus of mice, and influenza A virus differently affect respiratory allergy in mice
Authors: Barends M. , De Rond L. G. H. , Dormans J. , Van Oosten M. , Boelen A. , Neijens H. J. , Osterhaus A. D. M. E. , Kimman T. G.
Published: March 2004

Background
Respiratory viral infections in early childhood may interact with the immune system and modify allergen sensitization and/or allergic manifestations. In mice, respiratory syncytial virus (RSV) infection during allergic provocation aggravates the allergic T helper (Th) 2 immune response, characterized by the production of IL‐4, IL‐5, and IL‐13, and inflammatory infiltrates. However, it is unclear whether the RSV‐enhanced respiratory allergic response is a result of non‐specific virus‐induced damage of the lung, or virus‐specific immune responses.

Objective
In the present study we investigated whether RSV, pneumonia virus of mice (PVM) and influenza A virus similarly affect the allergic response.

Methods
BALB/c mice were sensitized and challenged with ovalbumin (OVA), and inoculated with virus during the challenge period. Pulmonary inflammation, lung cytokine mRNA responses, and IgE production in serum were assessed after the last OVA‐challenge.

Results
Like RSV, PVM enhanced the OVA‐induced pulmonary IL‐4, IL‐5, and IL‐13 mRNA expression, which was associated with enhanced perivascular inflammation. In addition, PVM increased the influx of eosinophils in lung tissue. In contrast, influenza virus decreased the Th2 cytokine mRNA expression in the lungs. However, like PVM, influenza virus enhanced the pulmonary eosinophilic infiltration in OVA‐allergic mice.

Conclusion
The Paramyxoviruses RSV and PVM both are able to enhance the allergic Th2cytokine response and perivascular inflammation in BALB/c mice, while the Orthomyxovirus influenza A is not.