bluuebunny/crossref_metadata_2025 huggingface dataset:
Excessively crying infant in the family: mother–infant, father–infant and mother–father interaction
Authors: Räihä H. , Lehtonen L. , Huhtala V. , Saleva K. , Korvenranta H.
Published: March 2003

Method

We studied mother–infant, father–infant and mother–father interaction in 32 families with an excessively crying infant and in 30 control families. The group with excessive criers was divided further into subgroups of severe colic (n = 13) and moderate colic (n = 19). The three dyads of the family were video‐recorded when the infants were an average of 5 weeks old. The assessment was carried out during the infant's feeding, nappy change and discussion between the parents. During the assessment, only four infants were crying. The Parent Child Early Relational Assessment Scale and the Beavers Scale were used.

Results

The main findings suggest that both parents of colicky infants had less optimal parent–child interaction compared with the control parents. The problems in the interaction were most pronounced between the fathers and infants in the severe colic group. The father–infant interaction was less optimal in 13 items of 65 (20%) in the severe colic group, in one item of 65 (2%) in the moderate colic group and in none of the items in the control group. The mother–infant interaction was less optimal in six items out of 65 (9%) in the severe colic group, in three items out of 65 (5%) in the moderate colic, and in none of the items in the control group. Severely colicky infants were also less competent in interacting with their parents. In addition, interaction between the parents was more often dysfunctional in the severe colic group.

Conclusions

The problems in early family interaction may threaten the well‐being of families with excessively crying infants and they therefore deserve special attention from the health care professionals.
Parents’ stories of sensitive and insensitive care by paediatricians in the time leading up to and including diagnostic disclosure of a life‐limiting condition in their child
Authors: Davies Ruth , Davis Bryn , Sibert Jo
Published: March 2003

This study explored parents’ experiences of care by paediatricians in the time leading up to and including diagnostic disclosure of a life‐limiting condition in their child.

Research methods   Qualitative exploration using in depth interviews with a purposive sample of 30 families whose child was diagnosed with a life‐limiting condition.

Results   Parents’ stories centred upon their need to have their concerns and needs as parents validated by paediatricians. Demonstrable qualitative differences were shown to exist between the sensitive and insensitive clinical practices of paediatricians. Parents’ stories identify the need for clinical practice to be based upon a humanistic approach to care. These also identify how sensitive paediatricians achieved this ideal through meeting the ‘double obligation’ of combining technical skills with an understanding of their needs as parents.
Clinical utility of serum levels of eosinophil cationic protein (ECP) for monitoring and predicting clinical course in childhood asthma
Authors: Fujisawa  , Terada  , Atsuta  , Iguchi  , Kamiya  , Sakurai
Published: March 2003

Background

The concentration of ECP in serum has been proposed as a marker of airway inflammation in asthma. However, its clinical significance is still to be determined.

Objectives

This study was performed to determine whether concentration of ECP in serum reflects clinical status in asthma and can serve as a predictive parameter.

Methods

Cross‐sectional analysis was performed in 28 children with asthma. A total of 91 blood samples was obtained to determine levels of ECP in serum and eosinophil counts. Forced expiratory volume in 1 s was also determined at the time of the sampling. Data were analysed on the basis of asthma symptoms in the 4 weeks before and the 4 weeks after sampling.

Results

Serum levels of ECP were significantly lower in patients who had been asymptomatic for 3 or 4 weeks before sampling than in patients who had been symptomatic or asymptomatic for only 1 or 2 weeks. In the former group, serum levels of ECP were higher when patients became symptomatic after sampling than when they remained stable, a finding that suggests that serum levels of ECP may have a predictive value in certain situations. Although the concentration of ECP in serum was not proved to be predictive in the latter symptomatic group, the concentration of ECP was significantly lower when measured again 4 weeks later when the patients' symptoms had resolved. In contrast, levels of ECP were unchanged when patients remained symptomatic, a finding that suggests serum levels of ECP may reflect the clinical response to therapy.

Conclusions

Serum ECP may be a useful marker for monitoring and predicting the clinical course in asthma.
Anaphylactic reactions associated with trimethoprim
Authors: Bijl  , Van Der Klauw  , Van Vliet  , Stricker
Published: March 2003

Background

Anaphylactic reactions to cotrimoxazole are often ascribed to the sulphamethoxazole component of this antibacterial drug.

Objective

To determine whether the trimethoprim component can be the cause of an anaphylactic reaction.

Methods

An analysis was made of reports on anaphylaxis attributed to trimethoprim, as notified to the Drug Safety Unit of the Dutch Inspectorate for Health Care.

Results

In the period between September 1981 and November 1995, 13 such reports were received. Nine were classified as probable anaphylaxis. Of these, the causal relationship between exposure to trimethoprim and anaphylaxis was classified as definite in three reports, and as probable in the other six. The remaining four reports were classified as possible anaphylaxis. In one of these, the causal relationship was classified as definite, and in three as probable.

Conclusion

Although anaphylaxis due to trimethoprim seems to be rare, it may be more common than previously thought. Apparently, anaphylaxis to cotrimoxazole is not always caused by sulphamethoxazole.
Indoor environment of residential homes in Hong Kong — relevance to asthma and allergic disease
Authors: Leung  , Lam  , Chan  , Lee  , Chan  , Pang  , Lai
Published: March 2003

Background

The prevalence of asthma and allergic disease has been increasing in Hong Kong and environmental factors are thought to play a major role.

Aim

To define the distribution of common inhaled allergens and air pollutants inside residential homes in Hong Kong.

Methods

Forty randomly selected residential homes were visited and surveyed. Dust samples were collected from mattress, bedroom floor, lounge room floor and kitchen floor for assays of Der p 1, Fel d 1, and Bla g 2. Nitrogen dioxide (NO2) levels were measured by stationary samplers left in the bedroom, lounge room and kitchen over a 1‐week period.

Result

All 40 homes were apartments in high‐rise buildings and the mean age of the homes was 7.6 ± 4.8 years. Visible dampness and mould patches were present in 27.5% of homes and the main fuel for cooking was gas (97.5%). Der p 1, Fel d 1 and Bla g 2 were detectable in at least 1 niche in over 85% of homes. In particular, all dust samples from mattress and bedroom floor contained Der p 1, and over 50% had levels ≥ 2 μg/g. The (geometric) mean Der p 1 level in mattress dust was 8.8 (0.3–157.8) μg/g. Fel d 1 and Bla g 2 were present in low levels throughout the homes with respective (geometric) means of 0.3 (0–3.7) μg/g and 0.1 (0–1.1) Unit/g in mattress dust. NO2 was readily detected in all niches but the highest level was in the kitchen with mean exposure of 48.7 ppb which was 1.5 times higher than that in the bedroom and lounge room.

Conclusion

Major allergens of mite, cat and cockroach, and NO2 are present in varying quantities in residential homes in Hong Kong. Exposure to these environmental factors could be important in the development of asthma and allergic diseases in susceptible individuals.
In vitro antigen‐specific sulphidoleukotriene production in patients allergic to Dermatophagoides pteronyssinus
Authors: Ferrer  , Sanz  , Prieto  , Oehling
Published: March 2003

Background

Sulphidoleukotrienes (slt) are important mediators in allergic diseases that are synthesized after allergen‐specific stimulation.

Objectives

The aim of this study is to determine in vitro slt production after allergen‐specific (Dermatophagoides pteronyssinus) stimulus of peripheral blood leucocytes and to observe whether histamine release in whole blood with the same allergen correlates with slt production. We also wanted to evaluate whether a correlation exists between the release of slt and histamine and other diagnostic procedures as well as various clinical situations.

Methods

We studied 62 patients sensitive to Dermatophagoides pteronyssinus (Der p), 30 atopic controls and 12 healthy donors. We determined slt production using the CAST‐ELISA technique and histamine release using two concentrations of Der p extract (20 and 2 ng/mL). We also carried out quantification of specific and total IgE levels, skin tests and pulmonary function test on each patient.

Results

We observed a significantly increased slt release after in vitro stimulation with Der p. There was a significant difference in the slt release between controls and sensitive patients (P < 0.001) and between atopic controls and sensitive patients (P < 0.001). The data are similar to those obtained with histamine release. We noted a positive correlation (P < 0.001) between slt and histamine release (r = 0.71, at 2 ng/mL and r = 0.83 at 20 ng/mL). We also found a positive (P < 0.001), although weak (r = 0.4 with at 2 ng/mL, and r = 0.34 with P = 0.003 at 20 ng/mL) correlation between slt release and specific IgE levels as well as between slt release and skin‐test reactivity (r = 0.49 at 2 ng/mL and r = 0.45 at 20 ng/mL; P < 0.001). No significant correlation between slt release and asthma severity was observed, although a trend toward higher slt production in severe and moderate asthma was detected. We found a significant (P < 0.001) but weak (r = − 0.3) negative correlation between age and slt release. With respect to sex‐related differences, we found significant differences (P < 0.05) in slt release between the sexes with a higher slt release in men than in women.

Conclusion

We conclude that CAST‐ELISA for quantification of slt production is a useful in vitro method for diagnosing sensitization to Der p. There also exists a close correlation between slt release and other parameters of allergic sensitization in vitro as well as in vivo.
Skin‐test reactivity and isotype‐specific immune responses to recombinant Asp f 3, a major allergen of Aspergillus fumigatus
Authors: Hemmann  , Ismail  , Blaser  , Menz  , Crameri
Published: March 2003

Background

The in vitro diagnostic value of recombinant allergens depends on the correlation between allergen specific IgE in serum and ability of the protein to elicit immediate type allergic reactions in sensitized individuals.

Objective

The study was carried out to evaluate the reliability of recombinant Asp f 3 (rAsp f 3)‐based serological determinations in ELISA and ImmunoCAP compared to skin‐test responses in A. fumigatus sensitized subjects.

Methods

Patients suffering from allergic bronchopulmonary aspergillosis (ABPA, n = 11) or allergic asthma with A. fumigatus sensitization (n = 8) and healthy control subjects (n = 4) were investigated. Intradermal skin tests (IDT) and allergen‐specific serology were performed using rAsp f 3 protein.

Results

All 11 patients with ABPA and five out of eight A. fumigatus‐sensitized asthmatics without ABPA exhibited a type I skin reaction. All rAsp f 3 skin test positive subjects showed relevant rAsp f 3‐specific serum IgE levels compared with IDT‐negative individuals who scored below the rAsp f 3‐ImmunoCAP cut‐off values of < 0.35 kUA/L. Analyses of rAsp f 3‐specific immunoglobulins revealed significantly higher serum levels of IgG, IgG1, IgG4 and IgE antibodies in patients with ABPA compared with A. fumigatus‐sensitized asthmatics and healthy controls.

Conclusion

rAsp f 3 represents a major allergen of A. fumigatus. It is recognized by 84% of asthmatic individuals sensitized to the fungus and elicits specific type I skin reactions. The skin test outcome strictly depends on the presence of rAsp f 3‐specific IgE and the lack of false positive or false negative results comparing skin test outcome with specific serum IgE levels determined by ELISA or rAsp f 3‐ImmunoCAP suggests the possibility to rely on serological data obtained with recombinant allergens to diagnose sensitization to A. fumigatus.
Oilseed rape allergy presented as occupational asthma in the grain industry
Authors: Suh  , Park  , Nahm  , Kim
Published: March 2003

Background

There have been several reports on respiratory allergic symptoms induced by pollen of oilseed rape. To the best of our knowledge, this is the first report dealing with oilseed rape dust mainly composed of seeds, as an occupational allergen in the grain industry. In this paper, we present a case of occupational asthma caused by oilseed rape dust from the Animal Feed Industry, which proved to be induced by an IgE‐mediated reaction.

Methods and results

The patient displayed positive respondes to Dermatophagoides farinae as well as oilseed rape dust extract. The bronchoprovocation test showed an early asthmatic response to oilseed rape dust extract. Serum specific IgE antibody to oilseed rape antigen was detected by enzyme‐linked immunosorbent assay (ELISA). ELISA inhibition test showed significant inhibitions with addition of oilseed rape antigen. In order to further identify the allergenic components of extract, sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and immunoblot analysis were performed. Fourteen IgE‐binding components ranging from 10 to 160 kDa were detected within the oilseed rape extract.

Conclusion

These results suggest that the inhalation of oilseed rape dust, not pollen, can cause IgE mediated bronchoconstriction in an exposed worker of the grain industry.
Allergenic relationship among four common and dominant airborne palm pollen grains from Eastern India
Authors: Chowdhury  , Chakraborty  , Gupta‐Bhattacharya  , Chanda
Published: March 2003

Background

Palm pollen grains are predominant aeroallergens in the tropics including India. Evidence of allergenic crossreactivity had been reported from various parts of the world on different families, e.g. Poaceae, Asteraceae, etc. No such information is available about the palm pollen of tropical countries.

Objectives

The present study was undertaken to find out the allergenic relationship, if any, in four common and important palm pollen in India.

Methods

A 2‐year aerobiological survey was carried out at Madhyamgram situated at the suburban fringe of Calcutta Metropolis using Burkard volumetric sampler to know the seasonal variation of Areca catechu, Borassus flabellifer, Cocos nucifera and Phoenix sylvestris among others. Skin‐prick tests (SPT) were performed with the relevant pollen extracts on the respiratory allergic patients. Sera from the subjects were tested directly by ELISA for estimating the allergen specific IgE. ELISA inhibitions and dot blotting were performed with pooled patients sera and four palm pollen extracts to detect the crossreactivity.

Results

Among 70 patients, Areca catechu exhibited the maximum percentage (48.5%) of positive responses followed by Cocos nucifera (45.7%), Phoenix sylvestris (42.85%) and Borassus flabellifer (38.5%). On an average, 30–50% of the patients showed positive skin reactions and enhanced specific IgE level to more than one palm pollen extract. Further evidence of allergenic crossreactivity among the four palm pollen grains were provided by dot blotting and ELISA inhibition studies. In ELISA inhibition, a distinct inhibition was obtained with comparable amounts of the pollen extracts.

Conclusion

The suburban aerobiological survey of Calcutta and SPT results confirmed that the relevant pollen types are significant contributors of aeroallergen load of the study area. ELISA inhibition studies with the pooled patients sera using antihuman IgE probe revealed the presence of shared allergenic components among the four palm pollen grains.
Allergenic and antigenic activity of peptide fragments in a whey hydrolysate formula
Authors: Van Hoeyveld  , Escalona‐Monge  , DE Swert  , Stevens
Published: March 2003

Background

Milk hydrolysates, although frequently used as substitutes in cases of cow's milk allergy, show a reduced but never a complete abolishment of antigenicity and allergenicity.

Objective

Our purpose was to determine the lower molecular weight limit of peptides to elicit skin reactions and to bind IgE antibodies in vitro.

Methods

Using FPLC, an ultrafiltrated whey hydrolysate, was fractionated in different molecular weight fractions. Skin‐prick tests were performed with the hydrolysate and its fractions in five cow's milk allergic children, and RAST inhibition tests were done using the serum of these children.

Results

On the basis of the lowest extinction values between two peaks of the chromatogram, seven fractions with molecular weights between 15 000 and 125 Da were obtained. Peptides of > 2600 Da elicited a clearly positive skin reaction and inhibited IgE‐binding, while peptides of < 1400 Da did not give any positive skin reaction but were still able to inhibit to a small extent IgE‐binding to the hydrolysate.

Conclusion

Our findings suggest that for skin reactivity peptides of > 1400 Da are needed. The minimal molecular mass for IgE binding in vitro appears to be situated between 1400 and 970 Da. Such peptides might be used to develop a safe formula for patients reacting to milk hydrolysates or even for tolerance induction.
Mapping of T‐cell epitopes of Phl p 5: evidence for crossreacting and non‐crossreacting T‐cell epitopes within Phl p 5 isoallergens
Authors: MÜller  , Karamfilov  , Kahlert  , STÜwe  , Fahlbusch  , Cromwell  , Fiebig  , JÄger
Published: March 2003

Abstract

Background

Group 5 allergens represent major grass pollen allergens because of their high sensitization indices. The identification of T‐cell epitopes of these allergens is a prerequisite for the design of immunotherapeutic strategies based on peptide vaccination or modified allergens with conserved T‐cell epitopes.

Objective

This study was undertaken to determine T‐cell epitopes on Phl p 5 major pollen allergen of timothy grass (Phleum pratense).

Methods

T‐cell lines (TCLs) and T‐cell clones (TCCs), specific to Phl p 5, were established from the peripheral blood of 18 patients allergic to grass pollen. All TCCs were mapped for epitope specificities using 178 overlapping dodecapeptides representing the primary structures of two isoforms of Phl p 5 (Phl p 5a and Phl p 5b). Phenotype and cytokine production profiles of TCCs were tested. Selected TCCs were analysed for HLA class II restriction.

Results

A total of 82 TCCs were isolated. All TCCs displayed the helper cell (TH) phenotype. Their reactivity with two recombinant expressed isoforms of Phl p 5a and Phl p 5b was heterogeneous. The epitope specificity of the TCCs was then revealed. Nineteen T‐cell epitopes could be identified on Phl p 5. Eighty‐one percent of mapped TCCs recognized three T‐cell reactive regions on the Phl p 5 allergen. Some TCCs were reactive with isoepitopes presenting on Phl p 5a as well as Phl p 5b. Allergen‐specific stimulation induced a TH0‐like type of cytokine production in 25 of 50 TCCs. Almost all TCCs secreted high concentrations of interleukin‐13.

Conclusion

Phl p 5, a major grass pollen allergen, contains several T‐cell epitopes. Some epitope regions were recognized by several patients. Epitope recognition pattern could not be correlated with special HLA class II haplotypes. T‐cell stimulating isoepitopes were found at corresponding regions of Phl p 5a and Phl p 5b isoforms.
Mapping susceptibility genes for asthma and allergy
Authors: Bleecker
Published: March 2003

Using family data, linkage analysis has been performed to determine the location in the genome of susceptibility genes for allergy and asthma. It has now become clear that there are multiple regions of the genome that contain susceptibility genes for allergy and asthma. The results from two genome screen studies will be reviewed and compared with results from candidate gene approaches. Results from several studies show evidence for linkage to chromosomes 5, 6, 11, 12, 13 and 14 for atopy, asthma or a related phenotype such as total serum IgE levels. Many of these regions contain candidate genes involved in regulating processes that may be involved in the development or progression of allergy and asthma. Some susceptibility genes may affect the expression of these disorders while others may affect response to therapy. Susceptibility to developing allergy or asthma appears to be due to the interaction of multiple genes with the environment.
The role of IgE in asthma
Authors: Yssel  , Abbal  , Pène  , Bousquet
Published: March 2003

Asthma is a multifactorial and complex disease in which allergic factors and non‐allergic triggers interact and result in bronchial obstruction and inflammation. Allergenic sensitization is important in the development of asthma and, although links between inhalant allergy and asthma have been known for many years, they have recently been re‐emphasized. Indoor allergens are associated with asthma prelvalence, severity and exacerbations whereas outdoor allergens such as pollens are associated with exacerbations. Moreover, there is a link between total IgE and asthma which appears to be independent of allergen sensitization. One of the typical aspects of airway inflammation of asthma is the infiltration of the airway wall by T helper type 2 (Th2) cells. These cells are attracted to inflammatory sites by adhesion molecules and chemokines among which CCR3 and CXCR4 receptors appear to be of importance. Differentiation of B cells into IgE‐secreting plasma cells is a complex cascade of events in which cytokines play a crucial role. Both IL‐4 and IL‐13 are inducing IgE synthesis whereas IFN‐γ and IL‐12 are blocking IgE synthesis. IgE production by B cells not only requires the presence of IL‐4 or IL‐13, but also a physical interaction between T and B cells, involving a number of surface and adhesion molecules such as CD40‐CD40L and CD28/CD80. Production of TH2‐cytokines is not restricted to T cells as basophils and mast cells can produce them indicating that these cells may be of importance in the synthesis of IgE.
Lipid mediator pathways in the lung: leukotrienes as a new target for the treatment of asthma
Authors: Dahlén
Published: March 2003

This article summarizes recent evidence supporting that antileukotriene drugs represent a new treatment of asthma which may be particularly effective when combined with drugs that have complementary effects on airway obstruction and inflammation.

 Firstly, it has been documented that glucocorticosteroids do not inhibit in vivo production of leukotrienes in asthmatics. In line with such findings, addition of antileukotriene drugs to a group of aspirin‐intolerant asthmatics maintained on conventional therapy was found to result in an improvement of the asthma over and above the effect of the baseline treatment with inhaled and/or oral glucocorticosteroids. Likewise, in a 6‐week trial in a group of severe asthmatics, the asthma deterioration caused by a reduction of the dose of inhaled steroids by half, was prevented by addition of a leukotriene antagonist to the lowered dose of glucocorticosteroids. Current evidence therefore supports that antileukotriene drugs treat components of the pathophysiology which are left unaffected by treatment with glucocorticosteroid.

 Secondly, in experimental studies as well as in a recent allergen bronchoprovocation study in asthmatics, it has been found that the combination of antihistaminics with antileukotriene drugs will result in a profound inhibition of both the early and the late phase of allergen‐induced airway obstruction. It is hypothesized that such a combination may be useful against bronchoconstriction induced by other asthma trigger factors as well as in the treatment of asthma and rhinitits.
Asthma in older people
Authors: Dow
Published: March 2003

From the clinical perspective, asthma in the older patient may be difficult to diagnose because of the non‐specificity of presentation and the wide range of differential diagnoses. Prior to confirmation of asthma in the older patient, both respiratory and cardiac investigation may be necessary. Polypharmacy is inherent in treating older people and accurate drug histories are essential in order to identify patients with drug‐induced bronchospasm and avoid interactions with asthma medication. Patient and carer education is vital, and a structured approach to follow‐up which includes measurement of lung function and assessment of inhaler technique should be carried out in every asthmatic regardless of age. Finally, there is a great need for health services and those in the commercial sector that are involved in asthma care to ensure that future research and development rises to the challenge of an ageing population.
Effects of erythromycin on experimental extrinsic allergic alveolitis
Authors: Miyajima  , Suga  , Nakagawa  , Ito  , Ando
Published: March 2003

Background

Recent clinical studies have demonstrated the efficacy of erythromycin for treating patients with chronic lower respiratory tract inflammation. Mechanisms related to the anti‐inflammatory action are yet to be determined.

Objectives

The therapeutic efficacy of erythromycin in experimental extrinsic allergic alveolitis (EAA) was evaluated.

Methods

A murine model of EAA was developed by intratracheal inoculations with particulate Trichosporon mucoides followed by erythromycin or josamycin treatment. Cell populations, specific antibodies, chemotactic activities, TNF‐α, IL‐1β, MIP‐2 and KC of bronchoalveolar lavage fluid (BALF); histopathology of the lung and footpad reaction; myeloperoxidase of the whole lung; and immunohistochemistry of intercellular adhesion molecule‐1 (ICAM‐1), at 6 and 96 h after the challenge, were examined.

Results

There was a marked neutrophilic alveolitis and bronchiolitis at 6 h, and lymphocytic alveolitis and perivenule cuffing at 96 h after the challenge. Increase in total inflammatory cells and neutrophils in BALF at 6 h was significantly suppressed by pre‐treatment with 5 mg/kg/day of erythromycin intraperitoneally for 5 days (P < 0.01), with no apparent effect on specific antibodies, chemotactic activity or cytokines. Erythromycin also suppressed the Arthus‐type reaction in the footpad (P < 0.01). Histopathological studies revealed that erythromycin markedly decreased neutrophils in the lung and skin lesions and myeloperoxidase in the lung, simultaneously with inhibiting ICAM‐1 expression. The therapy has no remarkable effects on lymphocytes or 96 h response. Josamycin had no effects on the model.

Conclusions

The therapeutic dosage of erythromycin significantly suppressed acute neutrophil influx into the lung, intradermal Arthus reaction and the expression of ICAM‐1 in the lesions of experimental EAA. Erythromycin may be effective for treating subjects with acute EAA.
Increased positivity of skin test and allergenic stability of glycerinated soybean hull extracts
Authors: Morell  , Codina  , Rodrigo
Published: March 2003

Background

Results obtained from previous experiences with soybean hull antigens suggest that the addition of glycerine to the extract increases positivity of the skin test.

Objective

The purpose of this study was to investigate the effect of this glycerine addition and the influence of different storage temperatures on the potency of soybean hull extracts.

Methods

Twenty‐two asthmatic patients admitted to emergency rooms during one of the soybean‐related Barcelona asthma epidemics were evaluated 2 years after the last epidemic for: (1) sensitivity to prick test with glycerinated and non‐glycerinated extracts, both fresh and stored at 4 °C, − 20 °C and − 70 °C for 30 days, and (2) specific IgE to a fresh hull extract. All extracts were also studied by sodium dodecyl suphate‐polyacrylamide gel electrophoresis (SDS‐PAGE).

Results

No differences in positivities were detected for prick tests performed the same day of preparation between non‐glycerinated (5/22; 22.7%) and glycerinated (4/22; 18.2%) extracts. After 30 days storage, positivities of prick test with non‐glycerinated extracts stored at 4 °C did not significantly differ from the results obtained on the first day. However, prick tests performed with glycerinated extracts stored for 30 days showed increased positivity for the extracts stored at 4 °C (7/22; 31%) (P < 0.05) as well as for those stored at − 20 °C and − 70 °C (9/22; 40.9%) (P < 0.001). This latter percentage is similar to that of specific immunoglobulin E detected by radioimmunoassay (10/22; 45.4%). Also, for the glycerinated extracts only, the quantitative response showed an increase in mean diameter of the induration at 30 days at the three temperatures, even though owing to the great standard deviation and limited number of patients, it was only significant for the extract stored at − 20 °C (P < 0.01). SDS‐PAGE gel densitometry at 30 days demonstrated a loss of protein in the bands with a molecular weight higher than 66.2 kDa for non‐glycerinated extracts stored at 4 °C.

Conclusions

The results of this study confirm previous observations with soybean hull allergen extracts and indicate that (1) storage of soybean hull extracts with glycerine addition for 30 days at the concentration studied (1:100 w/v) determines a significant increase in the positivity of skin tests, and (2) glycerine addition stabilizes soybean hull extracts at any of the temperatures studied.
Identification of a highly promiscuous and an HLA allele‐specific T‐cell epitope in the birch major allergen Bet v 1: HLA restriction, epitope mapping and TCR sequence comparisons
Authors: FRIEDL‐HAJEK  , SPANGFORT  , SCHOU  , BREITENEDER  , YSSEL  , JOOST VAN NEERVEN
Published: March 2003

Background

Allergen‐specific CD4+ T cells play an important regulatory role in atopic allergy.

Objective

To investigate the human leucocyte antigen (HLA) restriction and T‐cell receptor (TCR) usage of allergen‐specific T‐cell clones (TCCs) that react with defined epitopes of Bet v 1, the major birch pollen allergen.

Methods

Five Bet v 1‐specific TCCs derived from two birch pollen‐allergic individuals and specific for Bet v 1, were epitope‐mapped with overlapping synthetic peptides. In addition, HLA‐restriction and TCR CDR3 sequences were determined.

Results

Three TCCs reacted with a Bet v 1 peptide containing amino acid residues 21–33 (BP21), the other two TCCs reacted with a minimal peptide comprising residues 37–45 (BP37). Studies using neutralizing anti‐HLA‐monoclonal antibodies and HLA‐typed APCs showed that the BP37‐specific TCCs were resticted by a HLA‐DQA1*0301/DQB1*0603 heterodimer. In contrast, BP21 was recognized in a highly promiscuous manner. TCCs recognizing this sequence were restricted by HLA‐DPB1*0201, a HLA‐DQA1*0201/DQB1*0201 heterodimer, or HLA‐DRB3*0101. Reverse transcription‐polymerase chain reaction with primers for all known TCRAV and TCRBV gene segments, followed by CDR3 region sequencing, revealed the usage of five different TCRAV and four different TCRBV gene segments by the TCCs, as well as diversity in the joining region. All BP21‐specific TCCs contained a negatively charged residue in their CDR3α regions, the CDR3β regions showed a high concentration of polar and OH‐group bearing residues. BP37‐specific TCCs shared the amino acid combination LY in the middle of their CDR3α regions, the CDR3β regions showed high concentration of OH‐group bearing or charged residues.

Conclusions

This study shows the existence of a highly promiscuous T‐cell epitope in Bet v 1. The presence of additional T‐cell epitopes in Bet v 1 may, however, hamper the clinical applicability of the epitope. Likewise, the diversity in TCR usage by T cells recognizing the epitope does not support the development of TCR‐directed immunotherapy for birch pollen allergy.
Urticaria from beer: an immediate hypersensitivity reaction due to a 10‐kDa protein derived from barley
Authors: Curioni  , Santucci  , Cristaudo  , Canistraci  , Pietravalle  , Simonato  , Giannattasio
Published: March 2003

Background

Urticaria from beer has been reported in atopic patients. In these subjects, the skin‐prick test positivity to and presence of specific serum immunoglobulin (Ig)E for barley malt, the basic ingredient used in brewing, suggested a type I hypersensitivity to barley component(s).

Objective

To identify the beer allergen(s) and to investigate the presence of related proteins in barley.

Methods

Three patients with urticaria from beer and other atopic people, some of them suffering from baker's asthma, were examined for both prick test sensitivity to and the occurrence of serum‐specific IgE for partially purified proteins from beer. Allergen identification in beer, malt and barley was performed by immunoblotting.

Results

Skin‐prick tests and detection of specific IgE by both solid‐phase (RAST) and liquid‐phase (AlaSTAT) assays demonstrated that the 5–20‐kDa beer protein fraction contained the allergen. Immunoblot analysis with sera of patients with urticaria from beer showed that IgE bound only the 10‐kDa protein band in beer and malt, whereas a main 16‐kDa protein was revealed in barley in addition to a very faint 10‐kDa band. With the serum of a patient suffering from baker's asthma no IgE binding bands were observed in beer, whereas specific IgE binding to several proteins, including a major 16‐kDa component, were detected for both malt and barley.

Conclusions

Urticaria from beer is an IgE‐mediated hypersensitivity reaction induced by a protein component of approximately 10 kDa deriving from barley. This allergen does not seem to be related to the major barley 16‐kDa allergen responsible for baker's asthma. Because of the severity of the allergic manifestations to beer we recommend testing atopic patients positive to malt/barley and/or who exhibit urticarial reactions after drinking beer for their sensitivity to this beverage.
Sputum cellular and cytokine responses to inhaled endothelin‐1 in asthma
Authors: Chalmers  , Macleod  , Thomson  , Little  , Patel  , Mcsharry  , Thomson
Published: March 2003

Background

Endothelin (ET)‐1 is a 21‐amino acid peptide which has potent bronchoconstrictor activity. Animal studies show elevation of ET‐1 during experimental airway inflammation, and inhibition of inflammation by endothelin‐antagonists, suggesting pro‐inflammatory activity for ET‐1.

Objective

We wanted to assess any acute influence that bronchoconstrictor doses of inhaled ET‐1 might have on cells, tumour necrosis factor (TNF)‐α, interleukin (IL)‐1β, nitrite (NO2) and albumin in induced sputum in asthma.

Methods

Bronchial challenge was performed using nebulized ET‐1 (nebulized dose range 0.96–15.36 nmol) and placebo in 10 adult asthmatic subjects in a randomized double‐blind placebo‐controlled cross‐over study. Sputum induction was performed 30 min and 4 h after placebo or ET‐1 bronchial challenge.

Results

All subjects experienced dose‐dependent bronchoconstriction to inhaled ET‐1 with a mean (range) PC15 forced expiratory volume in 1 s (FEV1) to ET‐1 of 9.45 (1.2–21.7) nmol. Comparing ET‐1 with placebo inhalation, there was no change in sputum differential cell counts, TNFα, IL‐1β, NO2 or albumin at 30 min or 4 h after inhalation, nor was there a difference in these parameters at 4 h compared with 30 min after ET‐1 inhalation. There was no fall in FEV1 at 4 h after ET‐1 inhalation, suggesting that ET‐1 inhalation is not associated with a late bronchoconstrictor response.

Conclusions

We conclude that inhaled ET‐1 does not appear to stimulate an acute inflammatory response in asthma as assessed by differential cell count, TNFα, IL‐1β, NO2 and albumin concentrations in induced sputum.
Endothelial cells upregulate eosinophil superoxide generation via VCAM‐1 expression
Authors: NAGATA  , SEDGWICK  , VRTIS  , BUSSE
Published: March 2003

Background

In vitro eosinophil (EOS) adhesion to recombinant human (rh)‐vascular cell adhesion molecule (VCAM)‐1 stimulates superoxide anion (O2−) generation and enhances formyl‐methionyl‐leucyl phenylalanine (FMLP)‐activated O2− generation. Therefore, EOS adhesion via VLA‐4 to VCAM‐1 expressed on endothelium may be instrumental in the selective recruitment and function of EOS in airway inflammation.

Objective

We hypothesized that EOS interaction with endothelial cells expressing VCAM‐1 will undergo an enhancement in inflammatory function.

Methods

To determine this possibility, human umbilical vein endothelial cells (HUVEC) were stimulated with either a combination of interleukin (IL)‐4 and tumour necrosis factor (TNF)‐α (100 p M) or medium alone for 24 h; the expression of adhesion proteins on HUVEC and their effect on EOS O2− generation was subsequently determined.

Results

As determined by both enzyme‐linked immunosorbent assay and flow cytometry, IL‐4 and TNFα acted synergistically to induce VCAM‐1 expression on HUVEC. Treating HUVEC with IL‐4/TNFα also increased EOS adhesion and primed subsequent FMLP (0.1 μM) activated EOS O2− generation. Although EOS adhesion was partially inhibited by both antiα4 and antiβ2 monoclonal antibodies (MoAbs), O2− generation was completely inhibited by either antiα4 integrin MoAb (HP1/2) or anti‐VCAM MoAb (BBIG‐V1). Furthermore, enhanced O2− generation, but not adhesion, associated with IL‐4 + TNFα‐treatment of HUVEC was inhibited when EOS were treated with the platelet activating factor (PAF)‐antagonist WEB 2086 (20 μM), thus suggesting an involvement of PAF in priming EOS. However, paraformaldehyde fixation of IL‐4/TFN‐α treated HUVEC did not significantly alter EOS function.

Conclusions

These results suggest EOS adhesion to endothelial cells via an VLA‐4/VCAM–1 interaction may be important in the development of the function of this cell. Furthermore, our results suggest that modulation of EOS function involves two priming factors: EOS adhesion to HUVEC expressing VCAM‐1 and PAF.
Functional interleukin‐5 activity in peripheral blood mononuclear cells from adolescents with mite antigen asthma in remission
Authors: NOMA  , HAYASHI  , KAWANO  , YOSHIZAWA  , ISHIKAWA  , SAEKI  , AOKI  , MATSUURA
Published: March 2003

Background

Asthma improves in most children during adolescence such that a small minority of patients exhibit clinically significant symptoms by the age of 20 years.

Objective and methods

To investigate late allergic reactions, including eosinophil inflammation, associated with outgrowing mite antigen‐induced bronchial asthma during adolescence, the relationship between clinical status and functional activity of interleukin (IL)‐5 produced by Dermatophagoides farinae (Df)‐stimulated peripheral mononuclear cells (PBMCs) in culture was assessed in mouse IL‐3‐dependent cells transfected with the human IL‐5 receptor gene.

Results

Activity of IL‐5 spontaneously produced by PBMCs from either patients with mite‐sensitive bronchial asthma or nonatopic control subjects was low. The activity of IL‐5 produced by PBMCs stimulated with concanavalin A was significantly higher. Upon challenge with specific allergens, such as Df antigen, but not with irrelevant antigens, including ovalbumin, the in vitro activity was increased in patients with active disease and decreased in patients in remission.

Conclusion

Results suggest that the antigen‐specific up‐regulation of functional IL‐5 activity in late allergic reactions is reduced in patients in remission and likely to result in an improvement in clinical status. The Df antigen may suppress Df‐induced responses in patients with asthma in remission.
Elevated cord serum IgE increases the risk of aeroallergen sensitization without increasing respiratory allergic symptoms in early childhood
Authors: Tariq  , Arshad  , Matthews  , Hakim
Published: March 2003

Background

Increasing prevalence of allergic disorders has focused attention on primary prevention. There is a need to improve the accuracy of early‐life predictors of atopy so that the at‐risk population can be accurately defined and preventive measures instituted.

Objective

The predictive capacity of elevated cord IgE, with or without family history of atopy, to allergic symptoms and skin prick test (SPT) sensitization is evaluated in a birth cohort followed up prospectively for 4 years.

Methods

A birth cohort of 1456 consecutively born children was recruited in 1989. Data were collected on family history of atopy and cord serum total IgE (cord IgE) was measured. Of these, 1218 children were seen in the clinic at 4 years to determine the development of symptoms and signs of allergic disease and 981 were skin tested to a range of common food and aeroallergens.

Results

Of 1218 children reviewed at age 4 years, 218 (17.8%) had symptoms of respiratory allergy and, of those skin tested (n = 981), 192 (19.6%) reacted positively. Twice as many children with elevated cord IgE (≥ 0.5 kU/L) at birth became sensitized to aeroallergens by age 4 years (34.8% vs 17.3%, P < 0.001). Positive predictive value (PPV) of elevated cord IgE for the development of aeroallergen sensitization was better than that of family history of atopy (34.8 vs 22.6%). Combining paternal atopy with elevated cord IgE substantially increased the predictive capacity (PPV 77.8%). Cord IgE levels did not correlate with clinical asthma or rhinitis at age 4 years and PPV for allergic respiratory symptoms remained poor at all cutoffs.

Conclusion

Cord IgE is better than family history for predicting atopy as defined by allergen sensitization and this predictive value can be further increased by combining cord IgE with paternal atopy.
Allergy to goat and sheep cheese with good tolerance to cow cheese
Authors: Umpiérrez  , Quirce  , Marañón  , Cuesta  , García‐Villamuza  , Lahoz  , Sastre
Published: March 2003

Background

We report on a patient who experienced allergic reactions after eating goat cheese and after touching goat and sheep cheese, but not after consuming cow's milk dairy products.

Objective

To assess the allergenicity and IgE‐binding capacity of the caseins from the three different species.

Methods

Skin prick tests were carried out using whole milk and caseins from three different species (goat, sheep and cow), and whey fractions of cow's milk. Total serum IgE and specific IgE to cow's milk proteins were measured by CAP system and specific IgE against caseins and whole milk were determined by ELISA technique. To evaluate allergenic cross‐reactivity, inhibition of the IgE ELISA activity to goat's milk and goat casein was tested for the three caseins. SDS‐PAGE and immunoblotting was used to determine IgE binding bands in caseins.

Results

Skin tests were positive to sheep and goat's milk, sheep and goat casein, as well as to sheep and goat cheese. Total serum IgE was 66 kU/L and IgE determinations by CAP were negative. IgE ELISA against the caseins from goat and sheep was strongly positive, whereas it was negative to cow casein. ELISA inhibition assays revealed a high degree of cross‐reactivity between goat casein and sheep casein. Immunoblotting showed three IgE‐binding bands in goat casein at 31, 27 and 22 kDa, which may correspond to α‐, β‐ and γ‐caseins. A band at about 31 kDa was observed in sheep casein and another band at 34 kDa was recognized in cow casein.

Conclusion

This patient developed allergy to goat and sheep cheese with good tolerance to cow's milk. We identified goat casein as the main allergen causing sensitization in this patient as demonstrated by in vivo and in vitro tests. A high degree of cross‐reactivity between goat and sheep casein was observed.
Could the airway epithelium play an important role in mucosal immunoglobulin A production?
Authors: Salvi S. , Holgate S. T.
Published: March 2003

Immunoglobulin (Ig) A is the major immunoglobulin of the healthy respiratory tract and is thought to be the most important immunoglobulin for lung defence. The basis for the preferential generation of IgA‐secreting cells in the airway mucosa remains unclear. Given the half‐life of 5 days for the majority of IgA plasma cells, many IgA plasma cells must develop daily from B cells to guarantee a continuous supply of IgA antibodies in the airway mucosa. For this, the surrounding cells must provide a constant supply of cytokines necessary for B‐cell isotype switch, growth and differentiation into IgA‐secreting plasma cells. Studies with CD4+ T‐cell knockout mice, T‐cell receptor knockout mice and mice made transgenic for CTLA4‐Ig demonstrate normal mucosal IgA isotype switch, differentiation and IgA production, thereby suggesting that T cells are not critical for mucosal IgA production, and that other cell sources may be more important. Also, the bronchus‐associated lymphoid tissue (BALT), which is believed to be the major site where IgA isotype switch and differentiation of B cells into plasma cells occur with the help of cytokines released by T cells, is not a constitutive feature of the normal human lung. This indicates that other parts of the respiratory tract must carry out the BALT function. We have recently demonstrated that healthy human airway epithelial cells constitutively produce IL‐5, a major cytokine implicated in the growth and differentiation of post‐switch mIgA+ B cells to IgA‐producing plasma cells. Several studies have recently reported that the human airway epithelium also constitutively produces IL‐2, TGFβ, IL‐6 and IL‐10, factors which are essential for B‐cell clonal proliferation, IgA isotype switch and differentiation into IgA‐producing plasma cells. The close proximity of B cells to the airway epithelium probably ensures a constant supply of growth and differentiation factors necessary for mucosal IgA production. In addition, the epithelial cells produce a glycoprotein, called the secretory component, which not only confers increased stability to S‐IgA, but is also quantitatively the most important receptor of the mucosal immune system, since it is responsible for the external transport of locally produced polymeric IgA and IgM. Recent studies also suggest a possible role for epithelial cells in antigen presentation. Dendritic cells situated within the airway epithelium could directly present antigens to B cells and direct their isotype switch towards IgA1 and IgA2 with the help of cytokines produced by epithelial cells. Airway epithelial cells could therefore play a major role in the production of mucosal IgA antibodies which are essential for airway mucosal defence.