bluuebunny/crossref_metadata_2025 huggingface dataset:
Silverfish protein in house dust in relation to mite and total arthropod level
Authors: WITTEMAN A. M. , VOORNEMAN R. , VAN DEN OUDENRIJN S. , VAN LEEUWEN J. , AKKERDAAS J. , VAN DER ZEE J. S. , AALBERSE R. C.
Published: April 2006

Two assays have been developed to measure arthropod levels in house dust. The first assay measures silverfish antigens. The second assay measures invertebrate tropo‐myosin and gives a global assessment of the level of atthropod‐derived material. These assays and a Der p 1 and Der p 2 assay were used to analyse 53 dust samples. In most dust samples the ratio of tropomyosin/Der p 2 was higher than in mite body extract, indicating that the assay measures other arthropods besides mites. Silverfish antigen was detectable in most of the dust samples. In many homes in which the inhabitants were unaware of the presence of silverfish, silverfish antigen was detectable. Therefore for information on exposure an immunochemical analysis is superior to a questionnaire.
Topical ocular levocabastine reduces ICAM‐1 expression on epithelial cells both in vivo and in vitro
Authors: BUSCAGLIA S. , PAOLIERI F. , CATRULLO A. , FIORINO N. , RICCIO A. M. , PESCE G. , MONTAGNA P. , BAGNASCO M. , CIPRANDI G. , CANONICA G. W.
Published: April 2006

Background
Levocabastine is a selective topical H1 antagonist, effective in the treatment of seasonal allergic rhinitis and conjunctivitis.

Objective
We evaluated the possible effects of levocabastine eye drops on early (EPR) and late phase (LPR) inflammatory changes induced by allergen‐specific conjunctival challenge (ASCC). We focused our attention on the possible effect of levocubastine on expression of the intracellular adhesion molecule‐1 (ICAM‐1) on epithelial cells. Such a phenomenon is likely to play an important role in allergic inflammation.

Methods
The study was a double‐blind, placebo‐controlled, randomized trial, performed in cross‐over, outside the pollen season. Ten out‐patients suffcring from allergic rhinoconjunctivitis due to Parietaria Judaica (wall parietary) were enrolled. Patients randomly received levocabastine eye drops (0.5 mg/mL) or placebo eyedrops, one drop in the left eye (right eye as control), 30 min before ASCC. Clinical evaluation (hyperaemia, burning‐itching, lacrimation and eyelid swelling) and cytological assessment (number of neutrophils, eosinophils and lymphocytes, and ICAM‐1 expression on conjunctival epithelium) were evaluated at baseline, 30min and 6h after ASCC. In parallel, we evaluated the in vitro effect of levocabastine at concentrations ranging from 2 × 10−9 M to 2 × 10−5M on ICAM‐l expression and shedding by a continuously cultured differentiated epithelial cell line (WK).

Results
Levocabastine reduced symptom scores during EPR (15min and 30min, P < 0.002), inflammatory cell infiltration duritig FPR (P < 0.002 for neutrophils, eosinophils and lymphocytes) and ICAM‐1 expression on epithelium both during EPR (P < 0.002) and LPR (P < 0.02). LPR symptom scores and inflammatory cell infiltration were only slightly modified by levocabastine treatment. In vitro levocabastine at 2 ± 10−5 M concentration was able to down‐regulate basal ICAM‐1 expression, although it exerted no effect on ICAM‐1 expression by interferon‐γ (IFNγ)‐stimulated WK cells and on soluble ICAM‐1 release by epithelium.

Conclusion
Levocabastine exerts anti‐allcrgJc activity, in that it reduces in vivo inflammatory cell infiltration due to ASCC, and also adhesion molecule expression on conjunctival epithelium. The latter phenomenon may be due, at least in part, to a direct effect of levocabastine on epithelial cells.
Pathological changes according to the severity of asthma
Authors: CHO S. H. , SEO J. Y. , CHOI D. C. , YOON H. J. , CHO Y. J. , MIN K. U. , LEE G. K. , SEO J. W. , KIM Y. Y.
Published: April 2006

BackgroundThere have been many studies concerning pathological changes in bronchial mucosa from asthmatics; however, few studies has been carried out to evaluate pathological changes according to the severity of asthma.

ObjectiveThis study was designed to evaluate the cellular components in bronchoalveolar lavage fluid (BALF) and histologicai abnormalities in asthmatics according to the severity ot asthma.

MethodsBronchoalveolar lavages, bronchoscopic biopsies and ultrastructural examinations were performed in 13 asthmatics and 11 (BAL) or four (biopsies) non‐asthmatic controls. The proportions of epithelial cells and eosinophils in BALF were significantly increased in asthmatics and showed significant correlations with PC20Methwhich reflects bronchial hyperresponsiveness. Light microscopic examination revealed loss of epithelium, inllammatory cell infiltrations and thickening of the basement membrane which also showed significant correlation with PC20Meth. Hypertrophy of airway smooth muscles and hyperplasia of mucous glands were prominent in asthmatics but there was no difference according to the severity of asthma. Ultra‐structural examination revealed that basement membrane thickening on light microscopic examination is due to the increased subepithelial collagen deposition with normal thickeness of basal lamina.

ConclusionThese data suggest that loss of epithelial cells, infiltration of inflammatory cells, especially eosinophils, and increased deposition of subepithelial collagen play major roles in determining the severity of asthma and non‐specific bronchial hyperresponsiveness.
Influence of endogenous cortisol on eosinophil function in sensitized pigs: direct measurements of eosinophil peroxidase
Authors: FORNHEM C. , PETERSON C. G. B. , SCHEYNIUS A. , ALVING K.
Published: April 2006

Background and Objectives
Eosinophils are associated with bronchial asthma, but the role of the eosinophil is not fully understood. This study was initiated in order to study the influence of endogenous cortisol on eosinophil recruitment and activation in allergic inflammation in the lower airways in the pig.

Methods
Polyclonal and monoclonal antibodies against porcine eosinophil peroxidase (EPO) were raised. Detection of eosinophils in blood smears and lung biopsy specimens was achieved using the polyclonal antibody. For determination of porcine EPO in bronchoalveoiar lavage (BAL) fluid, a sandwich enzyme‐linked immuno‐sorbent assay (ELISA) with a detection limit of 0.15μig/L was developed. No cross‐reactivity with porcine myeloperoxidase was found. Pigs that had been actively sensitized with repeated subcutaneous injections of Ascaris suum antigen were acutely challenged with antigen in the lower airways under pentobarbitone anaesthesia.

Results
Control animals with plasma cortisol levels of approximately 400 nM did not exhibit infiltration of eosinophils into lung parenchyma or EPO‐release in the bronchial lumen within 8 h after challenge. However, in pigs treated with a cortisol‐synthesis inhibitor (metyrapone), resulting in plasma cortisol levels of approximately 40 nM, there was a marked eosinophil infiltration into lung tissue at 8 h. Furthermore, EPO levels in BAL fluid were increased in some, although not all, low‐cortisol animals. There was no infiltration of eosinophils into skin tissue in these animals.

Conclusions
It is concluded that, after allergen challenge in the lower airways of metyrapone‐treated pigs, newly recruited eosinophils infiltrate lung tissue specifically. Furthermore, a cortisol‐sensitive release of the eosinophil‐derived cationic protein EPO, into the bronchial lumen was established. This is, to our knowledge, the first description of direct measurements of the release of an eosinophil granule protein in a large animal model of allergy.
Airborne dust antigen exposure and specific IgG response in the potato processing industry
Authors: ZOCK J.‐P. , DOEKES G. , HEEDERIK D. , VAN ZUYLEN M. , WIELAARD P.
Published: April 2006

Background
High prevalences of work‐related respiratory symptoms in relation to organic dust exposure have been reported in the potato processing industry, but the responsible effect mechanisms are not known.

Objective
To study the possible role of a type III allergy in aetiology. Methods Specific immunoglobulin G (IgG) and IgG4 subclass antibodies against occupational airborne antigens were determined in sera from 131 potato processing workers and 36 non‐exposed controls. Personal exposure to airborne antigens was measured, and a preliminary biochemical characterization was carried out.

Results
Specific IgG was detectable in almost all sera, but levels were significantly (P<0.01) higher in potato processing workers compared with controls. Specific IgG4 was detectable in half of the workers' sera, but in none of the control sera. The antigens involved appeared to be heat‐labile potato proteins. Antibody levels increased during the processing campaign in most workers, and this increase was dependent on the level of antigen exposure. Both the difference in IgG titres between the occupationally exposed group and the non‐exposed group, and the exposure‐related increase in specific IgG titres seemed to be mainly due to specific antibodies of the IgG4 subclass. Specific antibodies showed a non‐significant tendency to lower levels in workers with work‐related respiratory symptoms.

Conclusion
Occupational respiratory exposure in the potato processing industry leads to a strong humoral immune response, most pronounced for lgG4 subclass antibodies. Type III allergy is, however, unlikely to play a predominant role in the aetiology of respiratory effects.
Antigen presenting cells in the nasal mucosa of patients with allergic rhinitis during allergen provocation
Authors: GODTHELP T. , FOKKENS W. J. , KLEINJAN A. , HOLM A. F. , MULDER P. G. H. , PRENS E. P. , RIJNTES E.
Published: April 2006

Background
The role of antigen presenting cells (APC) in allergic rhinitis is underexposed. Allergen presentation to T lymphocytes is probably an important aspect of the pathophysiological mechanism of allergic rhinitis.

Objectives
The aim of the study was to investigate the presence and dynamics of APC with special emphasis on Langcrhans cells (LC) in the nasal mucosa of patients with an isolated grass pollen allergy during an out‐of‐season 2‐week allergen exposure, mimicking the natural grass pollen season.

Methods
Seventeen patients with isolated grass pollen allergy and four control subjects were challenged daily with allergen during a 2‐week period in the winter. Biopsy specimens were obtained once before, six times during and once after the provocation period. Biopsy sections were stained with monoclonal antibodies: OKT6 (CDla‐Langerhans cells). Ki‐M6 (CD68 macrophages), L25 (dendritic cells), anti‐IgE, HLA‐DR and HLA‐DQ (Major Histoeompatibihty Complex Class II ‐ antigen presenting eells), as well as staining with acid phosphatase.

Results
APC with different characteristics are present in the epithelium and lamina propria of the nasal mucosa. The number of LC increased significantly in epithelium and lamina propria. IgE+‐LC were present in the nasal mucosa and increase during provocation, HLA‐DR+ cells with dendritic and lymphocytic morphology and HLA‐DQ+ cells were found. The number of these cells increased during provocation in epithelium and lamina propria. The number of HLA‐DR+ epithelial cells did not change. A significant increase in the number of Ki‐M6+ cells (macrophages) was found in the lamina propria. However, Ki‐M6+ cells increased to the same extent in the lamina propria in the control group.

Conclusion
APC are influenced by allergen provocation. This study supports the hypothesis that (IgE+) LC are involved in allergic rhinitis. The role macrophages play remains doubtful.
IL‐4 induced leucocyte trafficking in cynomolgus monkeys: correlation with expression of adhesion molecules and chemokine generation
Authors: GUNDEL R. , LINDELL D. , HARRIS P. , FOURNEL M. , JESMOK G. , GERRITSEN M. E.
Published: April 2006

Background
Interleukin‐4 (IL‐4) is an immunoregulatory cytokine which has a wide variety of effects on immune cell function. In addition, recent studies suggest that IL‐4 may have effects on other cells including endothelial cells in terms of the regulation of adhesion molecule expression and leucocyte extravasation from the vascular space to sites of tissue inflammation. Consequently, IL‐4 may have an important role in the pathogenesis of allergic inflammation and disease.

Objective
The purpose of this study was to learn more about the potential role of IL‐4 in inflammatory disease, specifically in regard to the potential of IL‐4 to induce the expression of adhesion molecules on vascular endothelial cells and promote the adherence and transmigration of circulating leucocytes to sites of tissue inflammation.

Methods
Single subcutaneous injections of human IL‐4 were administered to cynomolgus monkeys and tissue biopsy samples were obtained and analysed for adhesion molecule expression on vascular endothehum and inflammatory cell infiltrates. In another series of experiments, multiple subcutaneous injections of human IL‐4 were administered (bid on four consecutive days) and the effects on peripheral blood leucocytes and plasma levels of various cytokines and chemokines were examined. Results Intradermal injection of IL‐4 induced the expression of vascular cell adhesion molecule‐1 (VCAM‐l) on cutaneous vascular endothelium that was present at 8hr and persisted out to 24h post injection. The expression of VCAM‐1 was associated with an inflammatory cell infiltrate comprised of granulocytes and mononuclear cells. Multiple injections of IL‐4 resulted in a dose‐related decrease in the relative percentage and total number of circulating lymphocytes and an increase in circulating neutrophils (4.6 ± 1–2.1 ± 0.2 ± 106/mL and 1.7 ± 0.3–7.0 ± 1 ± 106/mL, respectively). Analysis of cell surface markers by flow cytometry revealed a transient decrease in the number of CD4+ T lymphocytes and a sustained decrease in CD16+ cells. In addition. IL‐4 administration resulted in a large increase in plasma MCP‐1 concentration.

Conclusion
This is the first study to demonstrate an acute eflect of IL‐4 consistent with lymphocyte trafficking out of the vascular space, the induction of VCAM‐l expression on vascular endothelium and increases in plasma levels of MCP‐1 in vivo. We suggest that IL‐4 may be involved in the early recruitment of mononuclear cells to sites of tissue inflammation by the upregulation of VCAM‐l expression on vascular endothelium and the generation and release of potent chemoattractants.
Analysis of sequence polymorphism of a major mite allergen, Der p 2
Authors: CHUA K.Y. , HUANG CH. , SHEN H.D. , THOMAS W.R.
Published: April 2006

Background
The major house dust mite allergen Der p 2 has been regarded as an important allergen involved in the immunopathogenesis of allergic asthma and eczema. Objectives To determine the degree of sequence polymorphism exists in Der p 2 at both the genomic DNA and cDNA levels.

Methods
Isolation of cDNA clones was performed by screening the cDNA libraries with Der p 2 cDNA probe and the genomic sequences for Der p 2 were obtained by PCR amplification from environmental dust mites with Der p 2‐specific primers. DNA sequencing was carried out by the dideoxynucleotide chain termination method. The study of Der p 2 isoforms was performed by 2‐D gel immunoblot analysis using mouse anti‐Der p 2 serum.

Results
In this study, we have characterized the seqtiences of three difierent genealleles coding for the major house dust mite allergen Der p 2 at the cDNA level. The translated polypeptides from these clones differed from each other by 3–4 amino‐acid residues. These polymorphic residues determined were also found in Der f 2 and they were located in regions containing T‐epitopes. In addition, the genomic DNA sequences of Der p 2 which were obtained by PCR amplification using the environmental mites isolated from Taiwan and Australia have helped to confirm the authenticity of the polymorphisms detected in the cDNA clones generated from CSL cultured mites. Furthermore, 2D‐ immunoblot analysis indicated that there were at least 10 different isoforms (p 1 values range from greater than 7.0–5.9) of Der p 2 proteins produced by CSL cultured mites.

Conclusion
The results showed that there was a small but significant degree of sequence polymorphisms exists in Der p 2 gene alleles. Interestingly, the polymorphic residues were found in regions containing previously determined T‐epitopes. The polymorphism data reported here will be important for the understanding of the immune responses of mite allergens as well as for the development of the peptide‐based immunotherapeutic reagents for mite allergy.
Relationship between nasal hyperreactivity, mediators and eosinophils in patients with perennial allergic rhinitis and controls
Authors: DE GRAAF‐INT VELD C. , GARRELDS I. M. , KOENDERS S. , GERTH VAN WIJK R.
Published: April 2006

Background
In perennial allergic rhinitis, patients are almost daily exposed to aeroallergens. This ongoing allergic reaction results in increased sensitivity to allergens and non‐specific stimuli. It is generally known that inflammatory cells and mediators are involved in the pathogenesis of the allergic reaction.

Objectives
To study the relationship between nasal hyperreactivity and nasal inflammation during natural allergen exposure.

Methods
In 48 patients with perennial allergic rhinitis and in 11 volunteers a nasal brush, a nasal lavage and a histamine challenge were performed. Nasal inflammation was estimated by the number of eosinophils, levels of albumin, tryptase, prostaglandin D2 (PGD2), eosinophil cationic protein (ECP) and leukotriene C4/D4/E4 (LTC4/D4/E4).

Results
In contrast to PGD2 and tryptase, eosinophils (1.9 vs 0%, P = 0.0023), LTC4/ D4/E4 (17.51 vs 1.43pg/mL, P= 0.0001) and albumin (8.61 vs 2.37mg/mL, P= 0.0008) were significantly increased in rhinitis patients as compared with controls. Patients also showed increased responses to nasal histamine challenge assessed using a composite symptom score (21.5 vs 4 points, P=0.0001). The nasal response to histamine was weakly correlated with the total number of eosinophils in the cytospin (correlation coefficient r=0.38, P= 0.009).

Conclusion
Nasal hyperreactivity is correlated with the percentage of eosinophils in patients with perennial rhinitis. The patients' mediator profiles suggest that eosinophils are important in the ongoing allergic reaction and nasal hyperreactivity.
Reduction in humidity as a method of controlling mites and mite allergens: the use of mechanical ventilation in British domestic dwellings
Authors: FLETCHER A. M. , PICKERING C. A. C. , CUSTOVIC A. , SIMPSON J. , KENNAUGH J. , WOODCOCK A.
Published: April 2006

Background
Low humidity is an important limiting factor for mite population growth. Reducing humidity can therefore be used as a method to control mites within the home.

Objective
Ihis study investigated the effect of mechanical ventilation heat recovery (MVHR) units on house dust mites and mite allergen Der p I in typical homes in the North‐West of England.

Methods
Mite counts and Der p 1 levels were measured at 3‐monthly intervals over a period of 1 year in 18 houses (nine with MVHR units and nine architecturally matched control houses). Paired dust samples were collected using a vacuum cleaner with an air‐flow rate 451/sec, adapted to collect the sample onto a preweighed filter paper. A 1 m2 area of bedroom carpet, living room carpet and mattress was sampled for 2min. Indoor temperature and relative humidity (RH) levels were recorded for a period of 1 week before and after the winter period (November and February: 3 and 6 months data sets). The environmental questionnaire was completed at the beginning and at the end of the study.

Results
No difference in either Der p 1 concentrations or mite counts in any of the sampling sites at 3, 6, 9 and 12 months as compared with the baseline values was found, both within and between the groups (P > 0.1). The measured levels of RH performed in autumn and winter were found to be lower in the MVHR houses compared to the architectural controls. The indoor temperature during each period did not differ between the groups. Questionnaire data showed that the severity of condensation improved in the MVHR homes, whilst during the winter period, the severity of condensation had increased in the architectural control group.

Conclusions
The MVHR unit does not reduce indoor humidity to levels capable of retarding mite population growth and decreasing mile allergens in the type of houses predominantly found in the mild and humid climate of the North‐West of England.
The rationale for use of topical corticosteroids in allergic rhinitis
Authors: MYGIND N. , DAHL R.
Published: April 2006

The rationale for using topical corticosteroids in the treatment of allergic rhinitis is that high drug concentrations can be achieved at receptor sites in the nasal mucosa, with minimal risk of systemic adverse effects. Topical corticosteroids have been demonstrated to reduce the number of Langerhans' cells (or their markers) in the nasal mucosa, and this is thought to attenuate antigen presentation. T lymphocytes have been identified as being significant in orchestrating the immune‐inflammatory response, particularly the TH2 cells, which represent an important target for topical corticosteroids. TH2 cell‐evoked mast cells and basophils are the sole producers of histamine, a mediator of major importance for rhinitis symptoms. Several studies have shown that the increased number of mast cells and basophils in the epithelium following antigen challenge/exposure, are markedly reduced by topical corticosteroids. Furthermore, the number of eosinophils, an important morphological marker of allergic rhinitis, can be profoundly reduced by treatment with topical corticosteroids. The rationale for topical treatment is strengthened by evidence of inhibition of cytokine release from surface epithelial cells, resulting in reduced recruitment and activation of mast cells, basophils, and eosinophils, which may be attributed to the high drug concentration achieved in epithelial cells. Ongoing inflammation in the mucous membrane is indicated by entry of plasma into the nasal lumen which subsides with the anti‐inflammatory efficacy of topical corticosteroids. In contrast to anlihistamine therapy, which has little effect on nasal blockage, pretreatment with topical corticosteroids results in almost complete attenuation of late‐phase symptoms including nasal blockage, and moderate efficacy in early phase symptoms. Clearly, the spectrum of anti‐inflammatory activity afforded by topical corticosteroid therapy is of clinical significance in reducing the three major symptoms of allergic rhinitis — sneezing, watery rhinorrhoea and nasal blockage.
Challenge tests in nose and bronchi: pharmacological modulation of rhinitis and asthma
Authors: MYGIND N. , DAHL R.
Published: April 2006

In order to study the pathophysiology of allergic airway disease and its response to pharmacotherapy, allergic and non‐allergic provocation challenge techniques can be employed. Lower airway challenge has been used widely, but the use of nasal challenge is becoming more widespread as its advantages are realized. New measurement techniques are also being used (e.g. acoustic rhinometry), along with more classical methods such as spirometry, peak airflow rate and symptom scores, to determine the response to challenge. In the lungs, allergen challenge produces a biphasic response, which is less clearly defined in the nose. Topical histamine challenge closely resembles the effects of an allergic reaction and acts by stimulating sensory nerve endings. Methacholine is also often used for nasal challenge (often in addition to histamine), due to its effects on glandular sensitivity. Exercise induces bronchoconstriction in asthmatics and can be imitated by inhalation of cold, dry air. Cold air induces glandular hypersecretion and nasal discharge in normal subjects, which is increased in severity in rhinitic patients. Drug effect investigations using antihistamines have shown that histamine is important in producing the symptom of sneezing, whereas nasal blockage is due to vasodilatation rather than plasma exudation and oedema. β2‐agonists reduce allergen‐induced symptoms by stabilizing mast cells, whereas cholinoceptor antagonists reduce watery nasal secretion. Increased responsiveness of sensory nerves and nasal glands is a characteristic clinical feature of asthma and rhinitis, which is responsible for the symptomatology. These effects can be reduced by topical corticosteroids.
Pathology of rhinitis and bronchial asthma
Authors: DJUKANOVIC R. , WILSON S. J. , HOWARTH P. H.
Published: April 2006

Studies of the pathology of rhinitis and asthma have identified similarities and differences between these two clinical conditions. With regard to symptoms, both the nose and the lower airways respond to neural stimulation by irritant substances, but a major difference is that engorgement of the capacitance vessels is the main cause of nasal obstruction in rhinitis, while muscle constriction is the major determinant of lower airway narrowing. There are also similarities and differences with respect to the role of infiammatory cells. In both conditions there is evidence of allergen‐induced mast cell activation, with production of an array of mediators (some mast cell‐derived and others originating from a variety of other cell types). Eosinophilia is also characteristic of both diseases — it is prominent even in mild forms of asthma, but is low in pollen‐sensitive rhinitics outside of the season. T‐cell activation and production of cytokines plays an important role in the development and maintenance of allergic disease, but the level of T‐cell activation may differ between asthma and rhinitis. Further research into differences in cellular activity and response to treatment between these two diseases may help define factors which will determine whether atopic disease is expressed in the upper, lower, or both parts of the respiratory tract.
Azelastine eye drops reduce and prevent allergic conjunctival reaction and exert anti‐allergic activity
Authors: CIPRANDI G. , BUSCAGLIA S. , CATRULLO A. , PESCE G. , FIORINO N. , MONTAGNA P. , BAGNASCO M. , CANONICA G. W.
Published: April 2006

Background Azelastine is a selective H1‐receptor antagonist, which has previously been demonstrated to be effective in the treatment of allergic rhinitis. We have recently demonstrated that nasal azelastine inhibits the clinical and intlammatory events following nasal allergen challenge. Particularly, we focused our attention on ICAM‐1 expression on epithelial cells, since it is the natural ligand of LFA‐1, an adhesion molecule expressed by leucocytes, including eosinophils.

Objective Since azelastine ocular drops are now available, the aim of the present study was the evaluation of the anti‐allergic activity in the model of allergen specific conjunctival challenge (ASCC).

Methods Twenty outpatients with allergic rhinoconjunctivitis due to Parietaria Judaica (Wall Parietary) were included outside the pollen season. The study was designed as randomized, placebo‐controlled, double‐blind and parallel group, developed in two parts. The fonner investigated the onset of effect of a single dose of azelastine eye drops administered 20min after clinical response due to ASCC. The latter evaluated the clinical and inflammatory parameters following ASCC after 7‐days treatment with azelastine. Clinical parameters (hyperaemia, itching, lacrimation and eyelid swelling) were evaluated at baseline, 5, 10, 20 and 30 min (i.e. early phase reaction‐EPR) and 6h (i.e. late phase reaction‐LPR) after ASCC. Cytological assessment (number of neutrophils, eosinophils. monocytes and lymphocytes) and ICAM‐1 expression on conjunctival epithelial cells were evaluated at baseline, 30 min (i.e. early phase reaction‐EPR) and 6h (i.e. late phase reaction‐LPR) after ASCC.

Results When administered 30 min after ASCC, azetastine produced a clinical effect ranging between 10 and 20 min after eye drops administration (P < 0.01). After 7 days of treatment, 30 min after ASCC, azelastine induced a reduction of symptom scores during EPR and LPR (P<0.01), a reduction of inflammatory cell infiltration during both EPR (P 0.01) and LPR (P 0.01), and a reduction of ICAM‐1 expression during EPR and LPR (both P 0.01). Placebo did not modify any of the studied parameters.

Conclusion Azelastine eye drops exert anti‐allergic activity, inducing a rapid improvement of clinical events when administered after ASCC, and reducing both sytiiptoms and cellular infiltration when administered before ASCC. Finally, azelastine down‐regulates ICAM‐1 expression on epithelial conjunctival cells, confirming the results obtained at nasal level.
Heterogeneous proteolytic specificity and activity of the house dust mite proteinase allergen Der p I
Authors: HEWITT C. R. A. , HORTON H. , JONES R. M. , PRITCHARD D. I.
Published: April 2006

Background Exposure of the skin or respiratory tract to proteinases is frequently associated with allergic sensitization. This is of particular significance in the domestic indoor environment where the proteolytic activity of Der p I, the group I allergen of the house dust mite Dermatophagoides pteronyssinus, may influence the allergenicity of mites. Using class‐specific proteinase inhibitors and active‐site affinity chromatography, we have previously shown that Der p I exhibits a mixed cysteine‐serine proteinase activity. Measurement of the amount of cleavage, however, did not determine whether the inhibitors used were targeting exactly the same proteolytic mechanism.

Objective To resolve this issue, we have examined whether the cleavage specificity of the cysteine and serine proteinase activities of Der p I was the same.

Methods HPLC and mass spectrometry were used to analyse and identify the products of a Der p I‐digested peptide substrate and thus identify the peptide bonds cleaved.

Results Der p I cleaves different peptide bonds, depending upon the class of proteolytic mechanism used. In the model peptide substrate insulin B chain, the cysteine and serine proteinase activities of Der p I showed preference for glutamic acid and arginine respectively in the P1 position.

Conclusion These data suggest the existence of more than one mechanistic form of the allergen immunologically identified as Der p I. If proteolytic activity is indeed a function of allergenicity, this information may have important implications for the pathogenicity of Der p I and the ability of innate antiproteinase defences in the respiratory tract to prevent immune sensitization.
The effect of anti‐IL‐4 monoclonal antibody, rapamycin and interferon‐γ on airway hyperreactivity to acetylcholine in mice
Authors: NAGAI H. , MAEDA Y. , TANAKA H.
Published: April 2006

Background The role of IgE in airway hyperreaetivity is obscure.

Objective In order to clarify the role of IgE in airway hyperreactivity, we investigated the effect of anti‐IL‐4 monoclonal antibody, rapamycin and interferon‐γ on the antigen‐induced IgE response, airway eosinophilia and hyperreactivity in mice.

Methods Mice were immunized with an antigen (ovalbumin; OA) at intervals of 12 days. OA was inhaled 10 days after the secondary immunization. Twenty‐four hours after the last inhalation, airway reactivity to acetylcholine was measured and bronchoalveolar lavage fluid (BALF) was obtained.

Results Three inhalations of antigen caused an increase in the number of eosinophils in bronchoalveolar lavage fluid (BALF) and in airway hyperreactivity to acetylcholine with a significant elevation of serum IgE level. Anti‐IL‐4 at a dose of 1000 μg/animal and rapamycin at doses between 0.1 and 1 mg/kg inhibited the IgE production, but did not affect the airway eosinophilia or hyperreactivity to acetylcholine. In contrast, IFN‐γ clearly inhibited the antigen‐induced airway eosinophilia and hyperreactivity, but did not affect the IgE antibody production.

Conclusion These results suggest that the inhibition of IgE production does not suppress the onset of airway hyperreactivity and eosinophilia in mice, and that IFN‐γ inhibits the antigen‐induced airway hyperreactivity, probably due to the inhibition of airway eosinophilia.
The expression of murine eutaneous late phase reaction requires both IgE antibodies and CD4 T eells
Authors: SAWADA K. , NAGAI H. , BASAKI Y. , YAMAYA H. , IKIZAWA K. , WATANABE M. , KOJIMA M. , MATSUURA N. , KINIWA M.
Published: April 2006

Background Exposure of atopic patients to a specific allergen evokes an immediate response which is followed, in many cases, by a late phase reaction (LPR) some hours later. Here we have examined the immunological mechanisms required for the expression of cutaneous LPR in mice.

Methods BALB/c mice were immunized by i.p. injection of ovalbumin (OVA) and alum actively or by i.v. injection of anti‐OVA IgE monoclonal antibody (mAb) passively. After challenge by intradermal injection of OVA into ears, the changes in ear thickness, the number of eosinophils, and the levels of IL4 and IFN‐γ protein at the site of antigen challenge were examined.

Results Actively immunized mice developed a biphasic response at the site of OVA injection, while mice passively immunized with IgE anti‐OVA mAb displayed a strong early response but no LPR. Cell transfer experiments using BALB/c nu/nu mice revealed that both OVA‐specific IgE mAb and OVA‐primed CD4 T cells were required to evoke LPR. Moreover, LPR was associated with increased levels of IL‐4 production concomitant with reduced IFN‐γ production and was abolished by pretreatment with anti‐IL‐4 neutralizing mAb.

Conclusion It is suggested that murine cutaneous LPR against OVA is a type 2 inflammatory response in which both IgH antibodies and CD4 T cells play an obligatory role.
Oral cow milk challenge abolishes antigen‐specific interferon‐γ production in the peripheral blood of children with atopic dermatitis and cow milk allergy
Authors: SÜTAS Y. , HURME M. , ISOLAURI E.
Published: April 2006

Background A major feature of atopic dermatitis (AD) is the propensity to generate IgE to environmental antigens. Despite extensive information on IgE dysregulation in AD, the nature of immune responses to ingested allergens is poorly characterized.

Objective To determine the clinical and immunological responses to food in AD patients.

Methods To characterize the type and timing of clinical reactions to oral cow milk, 83 AD patients aged 2 to 60 months were subjected to double‐blind placebo‐controlled food challenge (DBPCFC). IFN‐γ and IL‐4 production by their peripheral blood mononuclear cells (PBMC) was determined before and after DBPCFC.

Results Of 50 patients positive to DBPCFC, 46% manifested exanthematous‐type immediate‐onset reactions and 54% eczematous‐type late‐onset reactions. In either group, the production of lL‐4 and IFN‐γ by Concanavalin A‐stimulated FBMC was comparable before and after DBPCFC. For immediate‐reacting patients, the median IFN‐γ production by milk‐stimulated PBMC was 11.5 (4.2–17.2) pg/mL as against 2.3 (0.2–5.7) pg/mL by unstimulated PBMC, P = 0.0008 before DBPCFC, and 4.6 (2.8–10.3) pg/mL vs 4.2 (1.7–9.0) pg/mL, P = 0.40, correspondingly after DBPCFC.

Conclusion Before DBPCFC, immediate‐reacting but not late‐reacting patients were found to be capable of allergen‐specific IFN‐γ production in vitro, indicating the heterogeneity in AD patients. After DBPCFC, the IFN‐γ generation abolished, indicating the effect of oral allergen exposure on IFN‐γ‐producing responses of AD patients.
Mite allergy is associated with a specific profile of IgG epitopes recognized on antigen p1 of Dermatophagoides pteronyssinus
Authors: DUCHATEAU J. , MICHILS A. , MICHEL O. , BARAS L.
Published: April 2006

Background Immune response lo inhaled antigens differs in allergic patients and healthy individuals, mostly in the quality of T cell help provided (i.e. Th2 or Th l dominant subset). However, while different in many functional aspects, both groups of T cells shared the capacity to support the synthesis of antigen‐specific immunoglobulin G (IgG) antibodies detected in different amounts in the serutn of atopic and healthy individuals.

Objective The present study investigates whether these IgG responses display similar or different epitopic dominance in the mite sensitization model.

Methods Antibody specilicity was evaluated by comparing the IgG binding to native Der p1 (nDer p1) and its products of pepsin hydrolysis (dDer p1) in 56 mite‐allergic patients and 148 healthy individuals, including 24 mite‐sensitized individuals in a solid enzyme linked immunosorbent assay (ELISA). Antibody specificity was also studied in competitive ELISA using streptavidin biotin technology.

Results Mite‐allergic sera showed a higher degree of binding to nDer p1 than to dDer p1, whereas control sera and mite‐sensitized sera bound at a similar level to the two forms of the antigen. Allergic sera and control sera, including mite‐sensitized sera, showed distinct capacities to prevent the binding to nDer p1 of pooled IgG from each group as well as murine monoclonal antibodies specific to Der p1.

Conclusion The IgG response to Der p1 of mitc‐allergic patients differs from that of healthy controls and mite‐sensitized subjects, not only in its increased titres but also in its consistant pattern of modified specificity, displaying a marked preference lor conformational epitopes. Cross‐competition experiments confirm the clinically associated, restricted specificity, allowing almost complete discrimination between groups, particularly between mite‐sensitized and mite‐allergic subjects, which is currently impossible with routinely available assays.
Effect of surfactant inhalation on allergic bronchoconstriction in guinea pigs
Authors: KURASHIMA K. , FUJIMURA M. , TSUJIURA M. , MATSUDA T.
Published: April 2006

Background In the small airway, surfactant reduces surface tension, prevents liquid filling of bronchioles, thereby maintaining patency in the small airways. Recent reports demonstrated that surfactant dysfunction develops in experimental asthma in immunized guinea pigs. However, there are few reports concerning surfactant and lung function in an experimental asthma model.

Objective To examine whether inhaled surfactant improves lung mechanics in antigen‐induced bronchoconstriction in guinea pigs.

Method We developed a passively immunized guinea pig model for allergic bronchoconstriction induced by antigen inhalation. Using this model, we investigated the effect of inhaled exogenous surfactant, surfactant TA. on the airway opening pressure (Pao) after antigen challenge.

Results Aerosol antigen challenge produced a gradual and long‐lasting increase in Pao. Twenty minutes after antigen challenge, aerosolized surfactant TA, 20 mg/ml. was inhaled for 90 s, and it significantly reduced the Pao by 32.8% in 12 min, while a 10.2% reduction was observed in a control group in the same period. When surfactant TA was administered by 90‐s inhalation before antigen challenge, it inhibited the Pao increase in a dose‐dependent manner: mean inhibitory rates of Pao were 33.6% in surfactant TA 10 mg/ml and 61.9% in surfactant TA 20 mg/mI, respectively.

Conclusion Inhaled surfactant showed preventive and recovery effects on antigeninduced bronchoconstriction in an immunized guinea pig model.
The natural history of Apis‐specific IgG and IgG4 in beekeepers
Authors: GARCÍA‐ROBAINA J. C. , DE LA TORRE‐MORÍN F. , VAZQUEZ‐MONCHOLI C. , FIERRO J. , BONNET‐MORENO C.
Published: April 2006

Background Most published studies on changes of specific IgG or its subclasses as a response to stimuli by allergens have been performed on patients under immunotherapy. There are few reports analysing the response to immunoglobulins in patients exposed to allergens in their natural habitats.

Objective The aim of this work was to discover the natural history of Apis specific IgG (IgG‐ap) and IgG4 (IgG4‐ap) levels in beekeepers from the Canary Islands.

Methods We studied 242 beekeepers (Bks). We used a questionnaire and measured total IgE and seric Apis specific IgE (IgE‐ap), IgG‐ap and IgG4‐ap against Apis mellifera venom.

Results All Bks had IgG‐ap and IgG4‐ap. IgE‐ap was positive in 160 Bks (65.6 %), but only 92 (37.6 %) Bks were considered sensitized. IgG‐ap and IgG4‐ap showed significant correlation (r = 0,84); IgE did not correlate with IgG‐ap or IgG4‐ap. There was no seasonal variation in IgG‐ap or IgG4‐ap. The group of sensitized Bks had significantly lower IgG‐ap and IgG4‐ap levels (P < 0.05). The groups with longer beekeeping activity showed significantly higher levels of IgG‐ap and IgG4‐ap (P < 0.001). Bks with locals reactions had significantly higher IgG‐ap and IgG4‐ap than Bks who reported systemic reactions (P < 0.05).

Conclusion Our study showed that IgG‐ap and IgG4‐ap appear to increase in Bks, either according to their beekeeping experience or in subjects with local reactions after bee stings.
Topical azelastine has a 12‐hour duration of action as assessed by histamine challenge‐induced exudation of α2‐macroglobulin into human nasal airways
Authors: GREIFF L. , ANDERSSON M. , SVENSSON C. , PERSSON C. G. A
Published: April 2006

Background Oral anti‐histamine drugs are widely used in the treatment of seasonal allergic rhinitis. Recently, anti‐histamines have become available also for topical treatment

Objective The present study, involving healthy subjects, examined the effect of topical azelastine on iuminal entry of α2‐macroglobulin and symptoms evoked by repeat histamine challenges during 24 h. The effect was cotnpared to a clinical dose of the oral anti‐histamine cetirizine and to placebo treatments.

Methods Placebo and azelastine (0.254 mg per nasal cavity) were delivered as two consecutive actuations per nasal cavity using a nasal spray device. Oral placebo and cetirizine (10mg) were given as single doses in a placebo‐controlled (double‐dummy), double‐blind, and cross‐over design. Histamine‐challenges were given 1 h before treatment, and 1, 6. 9, 12. and 24 h after each treatment. The nasal mucosal surface was lavaged after each challenge. The lavage‐fiuid levels of α2‐macroglobulin were determined to assess mucosal exudation of bulk plasma, and nasal symptoms were scored.

Results Histamine (40–400 μg/mL) produced dose‐dependent exudation and symptoms. Compared between each treatment and placebo, azelastine and cetirizine reduced the 40 and/or 400μg/mL histamine‐induced mucosal exudation of plasma from 1–12 h after treatment. In addition, cetirizine reduced the 40μg/mL histamine‐induced mucosal exudation of plasma 24 h after treatment. Differences between the two treatments were not evident regarding nasal symptoms.

Conclusion Histamine challenge‐induced mucosal exudation of plasma appears to be a useful method for studies of the duration of action of antihistamines. We conclude that topical azelastine is suited for b.i.d. therapy and that neither the exudative process nor watery secretion may impede the efficacy or the duration of action of tbis nasal drug.
Expression of adhesion molecules on mononuclear cells from individuals with stable atopic asthma
Authors: TOMITA K. , TANIGAWA T. , YAJIMA H. , SANO H. , FUKUTANI K. , HITSUDA Y. , MATSUMOTO Y. , SASAKI T.
Published: April 2006

Activated mononuclear cells (lymphocytes and monocytes) (MNCs) circulate in peripheral blood and accumulate in the airways of individuals with steady‐state asthma. The expression of adhesion molecules on MNCs of 10 patients with steady‐state atopic asthma and 10 non‐atopic control subjects was measured by flow cytometry. The mean fluorescence intensity of CD23 (P= 0.0003), CDl la (P= 0.034), and very late antigen‐4 (VLA‐4) (P= 0.016) was increased on lymphocytes of asthmatic patients relative to those of controls. Although the expression of CD16 (P= 0.002) and CD23 (P= 0.002). which are associated with differentiation into macrophages, was increased on monocytes of patients relative to those of controls, monocyte expression of VLA‐4 (P= 0.006) and sialyl Lewis X (P= 0.005) was reduced. The concentration of interleukin‐4 (lL‐4) in serum was significantly higher in asthmatic patients than in normal subjects (P= 0.023), and a significant correlation was apparent between the serum concentration of lL‐4 and the expression of VLA‐4 on lymphocytes (p= 0.71, P= 0.03) or on monocytes (p=−0.72, P= 0.03) in asthmatic patients. Results suggest that lL‐4 may contribute to the priming of adhesion molecule expression on lymphocytes even in steady‐state conditions in individuals with chronic allergic inflammation.
Allergenic crossreactivity between Lepidoglyphus destructor and Blomia tropicalis
Authors: JOHANSSON E. , SCHMIDT M. , JOHANSSON S. G. O. , MACHADO L. , OLSSON S. , VAN HAGE‐HAMSTEN M.
Published: April 2006

Background In general, the non‐pyroglyphid mites Lepidoglyphus destructor and Blomia tropicalis show a different geographical distribution. Allergic sensitization to both species have been demonstrated in several investigations. However, whether this reflects crossreactivity or dual sensitization is so far not known.

Objective The aim of the study was to investigate the allergenicity and allergenic crossreactivity of L destructor and B. tropicalis using sera from Sweden and Brazil.

Objective Allergens in extracts of L. destructor and B. tropicalis were identified with SDS‐PAGE and immunoblotting and the crossreactivity was studied by an immunoblot inhibition method. In addition to mite extracts, a recombinant major allergen of L destructor, Lep d 2, was used.

Results The extract prepared from L. destructor contained 21 IgE‐binding components when using the Swedish or the Brazilian sera. A 15 kDa allergen was recognized by 85% of the Swedish sera and 78% of the Brazilian. The B. tropicalis extract exposed 23 IgE‐binding components when the Brazilian sera were used and 19 when the Swedish sera were used. A total of 83% of the Brazilian sera and 80% of the Swedish sera identified a 14.5 kDa allergen. The IgE response of the Swedish serum pool to 10 B. tropicalis allergens was inhibited by L. destructor extract. Likewise, the response of the Brazilian serum pool to four different L. destructor allergens was inhibited by B. tropicalis extract. The recombinant Lep d 2 allergen inhibited 33% of the IgE binding of the Swedish serum pool to the 14.5 kDa allergen in the B. tropicalis extract.

Conclusion crossreactivity with several proteins from L. destructor and B. tropicalis was demonstrated. The results suggest that a B. tropicalis 14.5 kDa allergen is antigenically crossreactive with recombinant L. destructor allergen Lep d 2.
Management of children with potential anaphylactic reactions in the community: a training package and proposal for good practice
Authors: VICKERS D. W. , MAYNARD L. , EWAN P. W.
Published: April 2006

Background
The increase in potentially dangerous food allergic reactions (e.g. to peanuts and nuts) in children had lead to an increase in the prescription of adrenaline, including adrenaline for self‐ or parent‐injection. This raises problems in the management of these children in schools. Our experience in response to this is described.

Objective
To develop a training package to ensure that each child can continue at school but receive optimum emergency treatment in a safe environment. To provide education, training and support to school staff, and to support parents.

Methods and Results
This paper describes a model of good practice in the support of children with potentially fatal anaphylactic reactions. The following points are emphasized. Children with a history of anaphylactic reactions should be expertly assessed prior to the prescription of adrenaline injection. A written treatment plan is essential. Parents need expert support in managing the child. Schools need training and support. Children should lead a normal life, and not be stigmatized or labelled by their problem. Close liaison between primary care, acute services, community services and education services are essential for the successful management of this problem. Data from a questionnare showed the training package was of benefit, with considerable reduction in anxiety in school staff and parents.

Conclusion
A broad training package is essential: training in administration of adrenaline is only one aspect, and emphasis is placed on education, recognition of allergic reactions and prevention. The presence of children in school needing access to adrenaline is no longer perceived as a problem in our area. We recommend other health authorities provide a similar service.