bluuebunny/crossref_metadata_2025 huggingface dataset:
Study of eosinophil‐endothelial adhesion, production of oxygen radicals and release of eosinophil cationic protein by peripheral blood eosinophils of patients with rheumatoid arthritis
Authors: MERTENS A. V. , DE CLERCK L. S. , MOENS M. M. , BRIDTS C. H. , STEVENS W. J.
Published: April 2006

Beside lymphocytes and neutrophils, eosinophils are also involved in the inflammatory reaction in rheumatoid arthritis (RA). In this study, adhesion characteristics of peripheral blood eosinophils were studied in 43 RA patients and 19 controls, together with the expression of the β2‐integrin Mac‐1 (CD11b/CD18). In addition, the production of oxygen radicals of isolated peripheral blood eosinophils and serum levels of eosinophil cationic protein (ECP) were measured in order to evaluate eosinophil activation. Adhesion of eosinophils to unstimulated human vascular endothelium was significantly higher in RA patients with active disease (n= 4) compared with controls (n= 14) (P < 0.005) and compared with patients with less active RA (n= 16) (P < 0.05). Nevertheless, the expression of the adhesion molecule Mac‐1 (CD11b/CD18) was not increased in RA patients. ECP levels were higher in RA patients with active disease (P < 0.01). Release of oxygen radicals in response to phorbol stimulation was significantly elevated in active RA compared with controls (P < 0.05) and to less active RA (P < 0.05). We conclude that eosinophils of RA patients, especially those with active disease, are activated or at least primed and are involved in the inflammatory process in RA, analogous to the inflammation in asthma. The higher adhesion to inflamed endothelium is indicative of a higher infiltration in the joints, where tissue damage can be caused by toxic oxygen radicals and by granular proteins, such as ECP.
Annual variations in grass pollen seasons in London 1961–1990: trends and forecast models
Authors: EMBERLIN J. , SAVAGE M. , JONES S.
Published: April 2006

The record of daily average grass pollen concentrations monitored at St Mary's Hospital, Paddington. London, U.K. since 1961 is the longest duration pollen data set in Europe. Analysis of the results identifies the trends and characteristics of grass pollen seasons over three decades. During this lime seasonal allergic rhinitis has increased significantly in Britain. The annual start dates, length of season and severity are examined in relation to the main meteorological variables of cumulated temperatures above 5‐5°C and precipitation measured at one site within London and two in the surrounding rural areas. Land‐use changes are also considered. Significant decreases have taken place in both the duration and severity of the seasons, particularly between the 1960s and the early 1970s but also through the last 20 yr. This is largely a result of a decrease in pollen abundance in the region. The decline in pollen counts has slowed in recent years due to the increase in flowering grasses caused by the seaside policy and by uncut verges. Grass pollen seasons have tended to start later over the last two decades, despite an increase in the cumulated temperature profiles during late winter and spring. Empirical models have been developed using multiple regressions to incorporate meteorological and pollen data for the last 20 yr in order to forecast the start dates. duration and severity of the grass pollen seasons. These models were applied successful using the data for 1991 and 1992. Predictions of the main characteristics on the pollen seasons can be obtained relatively early in the year through the use of these models by employing the monthly weather forecasts in conjunction with long‐term average weather profiles.
Molecular cloning of a house dust mite allergen with common antibody binding specificities with multiple components in mite extracts
Authors: SHEN H‐D. , CHUA K‐Y. , LIN K‐L. , HSIEH K‐H. , THOMAS W. R.
Published: April 2006

Plaque radio‐immuno assay has been used to isolate an FgE‐binding clone from a lambda gt11 library of Dermatophagoides pteronyssinus cDNA, The clone HD6 contained DNA encoding a 215 residue protein which contained a predicted 17 amino acid residue leader sequence, no cysteines and a single W‐glycosylation site. The 198 residue mature protein would have a predicted MW of 22 177 D. No homologues were found in searches of the data banks. Sera from 14/38 allergic children reacted strongly with the polypeptide produced by the clone (37%). Skin tests showed reactivity in 16/30 (53%) allergic patients and 0/10 of controls. Affinity purification of rabbit antibodies with the clone showed that antibodies to the polypeptide had specificities to multiple products in mite extracts corresponding to components of Mr 29, 27 and 24 K by Western blotting. Absorption studies of IgE in allergic serum indicated further entities at 13 and 11.5 kD, It is proposed to name this allergen Der p VII.
Histamine‐free diet: treatment of choice for histamine‐induced food intolerance and supporting treatment for chronical headaches
Authors: WANTKE F. , GÖTZ M. , JARISCH R.
Published: April 2006

Histamine‐induced food intolerance is not IgE‐mediated. Skin‐prick testing and specific IgE to food allergens are typically negative. Food rich in histamine or red wine may cause allergy‐like symptoms such as sneezing, flush, skin itching, diarrhoea and even shortness of breath. The suspected reason is a diminished histamine degradation based on a deficiency of diamine oxidase. As diamine oxidase cannot be supplemented, a histamine‐free diet was implemented to reduce histamine intake. Forty‐five patients with a history of suffering from intolerance to food or wine (n=17) and chronic headache (n= 28) were put on the diet over months to years. Fish, cheese, hard cured sausages, pickled cabbage and alcoholic beverages had to be avoided. Complaint intensity and drug‐use per week prior to and 4 weeks after a histamine‐free diet were compared. After 4 weeks on the diet 33/45 patients improved considerably (p < 0.01), eight of them had total remission. In 12/45 patients, however, no changes in symptoms were observed. Symptoms of food or wine intolerance significantly decreased (P < 0.02; treatment of choice), headaches decreased in frequency (P < 0.001), duration and intensity. After eating histamine‐rich food symptoms were reproducible and could be eliminated by anti‐histamines in most patients. These data indicate the role of histamine in food and wine intolerance and that histamine‐rich food causes a worsening of symptoms in patients suffering from chronic headaches. Results obtained support the hypothesis of a deficiency of diamine oxidase in patients with intolerance to food or wine.
Abnormalities in histamine pharmacodynamics in chronic urticaria
Authors: KANNY G. , MONERET‐VAUTRIN D. A. , SCHOHN H. , FELDMAN L. , MALLIE J. P. , GUEANT J. L.
Published: April 2006

Histamine plays a key role in the pathogenesis of chronic urticaria (CU). The authors of this paper have studied the effects of ingested histamine in 25 patients with CU. A 120 ing dose of histamine. well‐tolerated in the healthy subject, was instillated into the duodenum. Concomitantly. plasma histamine (H) levels and plasma and urinary methylhistamine (MH) levels were measured. Intraduodenal administration of histamine was responsible for the development of an attack of urticaria in 64% of patients, while control subjects were asymptomatic. Plasma histamine levels were significantly higher after digestive histamine challenge (DHC) in patients with CU compared with controls. An abnormal increase in plasma histamine was observed in 72% of them. Plasma MH exhibited the same kinetic behaviour with a usually delayed time‐pattern. Urinary MH concentration was higher in patients presenting with early‐onset urticaria during the first hour than in those with the late‐onset type between 1 and 12 hr after DHC. The coefficient of mcthylation (plasma MH/MH + H) was not significantly different in patients presenting with an attack of urticaria following DHC and in other subjects, Urinary excretion of MH and urinary flow increased significantly in patients presenting with an attack of urticaria following DHC which corresponds to increased absorption of histamine during the 5‐hr period following DHC and its role on excretion by the kidney via vasodilation which it induces. This study demonstrates the abnormal frequency of disturbances in the metabolism of exogenous histamine in patients with CU. Increased plasma H accounts for the abnormal passage of H across the intestinal barrier which can result either from intestinal hyperpermeability and/or a deficit in the enzymatic catabolism of histamine. The systems of methylation and urinary clearance of MH appear to be effective. It is thus postulated that there is a deficit in diamine oxidase (DAO) in the enterocyte. The lack of correlation between the kinetic behaviour of plasma H and the onset of urticaria draws attention to the extent of individual variability in skin reactivity to histamine.
Influence of allergen avoidance at high altitude on serum markers of eosinophil activation in children with allergic asthma
Authors: BONER A. L. , PERONI D. G. , PIACENTINI G. L. , VENGE P.
Published: April 2006

A cohort of 12 asthmatic children was followed over several months, during which they moved back and forth from an allergen‐free to an allergen‐rich environment at high and low altitude, respectively. The children were treated with non‐steroidal anti‐asthmatic drugs as clinically needed. Histamine PC20‐FEV1 was unaltered during the study period, whereas serum levels of eosinophil cationic protein (ECP) and eosinophil protein X (EPX) showed significant changes when the children were exposed to the offending allergens. The total IgE significantly increased during exposure. The serum levels of myeloperoxidase (MPO) as well as of chemotactic factors for both neutrophils and eosinophils were unaltered during allergen exposure. We conclude that the serum markers of eosinophil activity ECP and EPX are sensitive indices of allergen exposure in asthmatic atopic children.
The involvement of eosinophils in the patch test reaction to aeroallergens in atopic dermatitis: its relevance for the pathogenesis of atopic dermatitis
Authors: BRUIJNZEEL P. L. B. , KUIJPER P. H. M. , KAPP A. , WARRINGA R. A. J. , BETZ S. , BRUIJNZEEL‐KOOMEN C. A. F. M.
Published: April 2006

Atopic dermatitis (AD) is considered a T‐cell mediated disease. Activated T‐cells, mainly of the CD4‐subtype. are abundantly present in lesional AD skin. Although not many intact eosinophils are present, deposits of eosinophil derived major‐basic‐protein (MBP) and eosinophil‐cationic‐protein (ECP) suggest eosinophil involvement. After patch testing AD patients with aeroallergens, an eczematous reaction develops after 24 48 hr at the site of application. This patch test reaction shows macroscopic resemblance to lesional AD skin and does not take place in normal individuals, asthma and allergic rhinitis patients. Lymphocytes together with eosinophils infiltrate into the dennis 26 hr after allergen application. Twenty‐four to forty‐eight hours after patch testing, eosinophils are in an activated state since they release ECP (being EG2‐positive). At this point in time eosinophils have also infiltrated the epidermis. Here they are EG2‐negative. Forty‐eight to seventy‐two hours after patch testing the eczematous reaction decreases. This coincides with disappearance of eosinophils from both the dermis and the epidermis; then, a dendritic staining pattern can be observed in the epidermis with anti‐eosinophil peroxidase. Thus, eosinophils infiltrate the dermis and epidermis after patch testing AD patients with aeroallergens and release part of their granular constituents. Recent in vitro investigations revealed that eosinophils from the circulation of AD patients react more powerfully in in vitro test systems such as chemi luminescence, chemotaxis and endolhelial adherence and transmigration. 11 is very likely that this activated (= primed) state is caused by the influence of lymphocyte‐derived cytokines like IL‐3, IL‐5 and GM‐CSF, since activated lymphocytes in the circulation (and tissue) may release these cytokines. The primed state of the eosinophils may facilitate tissue inlillration. The subsequent activation of eosinophils within the tissue leading to mediator release and the function of these mediators need to be further elucidated. The close similarity between the cellular events after a patch test reaction to aeroallergens in AD patients and those present in lesional AD skin suggests that the patch test reaction may be a helpful in rim model to study the pathogenesis of AD. The prominent involvement of lymphocytes and eosinophils in this reaction also suggests some similarity with late phase reactions (LPR) observed in the skin after intracutaneous allergen challenge.
Investigation of possible adverse allergic reactions to mycoprotein (‘Quorn’)
Authors: TEE R. D. , GORDON D. J. , WELCH J. A. , TAYLOR A. J. NEWMAN
Published: April 2006

Mycoprotein (‘Quorn’) is a food produced for human consumption from Fusetrium graminearum. Crossreactivity studies showed that mycoprotein shared multiple common allergenic determinants with Aspergillus fumigates and Cladosporium herbarum and some with Alternaria alternata. There is. therefore, a potential for mould allergic patients to react adversely to inhaled or ingested mycoprotein. Mycoprotein RAST screening of mycoprotein production workers was made during a 2 year period. Two of the production workers had specific RAST binding ≥2% but none reported symptoms. Two of 10 patients referred to hospital following vomiting and diarrhoea after ingestion of mycoprotein had a mycoprotein skin‐prick test weal ≥2 mm but none had a significantly raised RAST. These largely negative results are important and reassuring because consumption of the product in the U.K. is now widespread and increasing.
Sensitization to a weed pollen in a non‐atopic subject during a clinical study. A case report
Authors: GARCÍA‐ORTEGA P. , MARTÍNEZ A. , MARTÍNEZ J.
Published: April 2006

The case of a non‐atopic subject developing occupational sensitization and allergic rhinitis to Mercurialis annua pollen after intense exposure is described. The subject was involved in a clinical study by handling flowering plants daily. Preventive measures should be taken even in non‐atopic subjects exposed temporarily to proteins or potentially allergenic material.
Mite fauna, Der p I, Der f I and Blomia tropicalis allergen levels in a tropical environment
Authors: FERNÁNDEZ‐CALDAS E. , PUERTA L. , MERCADO D. , LOCKEY R. F. , CARABALLO L. R.
Published: April 2006

Fifty dust samples were collected from the mattresses and bedroom floors of 25 subjects with allergic asthma in Cartagena, Colombia, in order to identify house dust mites and quantitate Der p I, Derf I and Blomia tropicalis allergens. The geometric mean of the total mite density per gram of dust was 418 (range, 40–2280). Twenty‐two samples (44%) had more than 500 mites and four, less than 100. B. tropicalis and Dermatopha‐goides pteronyssinus were found in 96% and 90% of the samples, accounting for 40.1% and 35.7% of the total mites, respectively. Cheyletus malaccensis, Chortoglyphus arcuatus, Pyroglyphus africanus, Orihatids, Grallacheles bakeri. Tarsonemus spp., Suidasia spp., Dermatophagoides farinae and unidentified mites accounted for the rest. The geometric mean of the total mites/gram of dust in mattresses (563.9) was significantly higher than in floor dust (309. 1), P < 0.01. Allergen concentrations and mite numbers were analysed by Spearman rank correlations: B. tropicalis mites vs B. tropicalis allergen, r= 0.54, P<0.001; D. pteronyssinus mites vs Der p I, r= 0.52, P< 0.001. A negative correlation was obtained between B. tropicalis mites and Der p I. Allergens derived from B. tropicalis and other domestic mite species may play an important role in sensitization and allergic symptoms in Cartagena, Colombia.
Allergens from birch pollen and pollen of the European chestnut share common epitopes
Authors: HIRSCHWEHR R. , JÄGER S. , HORAK F. , FERREIRA F. , VALENTA R. , EBNER C. , KRAFT D. , SCHEINER O.
Published: April 2006

Type I allergy to pollen of the European chestnut (Castanea sativa) represents a major cause of pollinosis in (sub) Mediterranean areas. Using sera from 14 patients with established allergy to pollen of the European chestnut, 13/14 sera (92%) showed IgE‐binding to a 22 kD protein, 2/14(14%) displayed additional binding to a 14 kD protein and 1/14 (7%) bound only to the 14 kD protein of European chestnut pollen extract. Two monoclonal mouse antibodies, BIP 1 and BIP 4, directed against different epitopes of Bet v I (the major birch pollen allergen), and a rabbit antibody to recombinant birch profilin (rBet v II) were used to characterize the proteins of the European chestnut pollen. The recombinant birch pollen allergens, r Bet v I and r Bet v II (profilin) were employed to show common allergenic structures on proteins from both birch and European chestnut pollen by IgE‐inhibition experiments. Despite the fact that the 22 kD protein displayed a higher molecular weight in comparison to the 17 kD major birch pollen allergen, Bet v I, we could demonstrate reactivity of both monoclonal antibodies, BIP 1 and BIP 4, with this protein. A complete inhibiton of IgE‐binding to this 22 kD protein was shown by pre‐incubating sera with purified recombinant Set r I. In addition, the 14kD protein could be identified by IgE‐inhibition studies with recombinant Bet v II and by using a rabbit anti‐profilin antibody as the profilin from pollen of the European chestnut.
IgE specific immune responses to an African grass (Kikuyu, Pennisetum clandestinum)
Authors: POTTER P. C. , MATHER S. , LOCKEY P. , AINSLIE G. , CADMAN A.
Published: April 2006

Kikuyu grass (Pennisetum clandeminum) is widely used as a grazing pasture in Africa and, although it is extensively cultivated as a lawn for sports fields and domestic gardens, its allergenicity has never been studied in vitro. Using an extract of Kikuyu grass pollen, polyacrylamide gel electrophoresis. Western blotting and a monoclonal anti‐human IgE antibody, the specific IgE binding, in the serum of 160 allergic and non‐allergic patients in the Cape Town area, to a Kikuyu grass extract was studied. IgE in the sera of 43/104 known grass‐allergic individuals hound to Kikuyu grass on the Western blots. In addition, 4/28 ‘non‐allergic’ control subjects were found to have Kikuyu grass‐specific IgE. Five different profiles of specific IgE reactivity to Kikuyu grass antigens were oberved. In 29/43 patients, IgE bound to two dominant 48 and 70 kD allergens in the Kikuyu extract. Although a degree of cross‐reactivity with Bermuda grass (Cynodon dactylis) was found in immunoabsorption studies, the 48 and 70 kD allergens appear to be unique to Kikuyu grass. Exposure of heparinized blood from Kikuyu grass‐positive patients to the Kikuyu extract stimulated the release of histamine from their basophils in vitro. Kikuyu grass pollen is thus identified as an important aero‐allergen in South Africa.
Gastrointestinal stability of baker's yeast allergens: an in vitro study
Authors: KORTEKANGAS‐SAVOLAINEN O. , SAVOLAINEN J. , EINARSSON R.
Published: April 2006

An in vitro model was established to study the stability of baker's yeast (Saccharomvces cerevisiae) allergens in conditions simulating the gastrointestinal tract. The protocol consisted of 2 hr incubation under gastric conditions (pH 1.2, +37°C and gastric enzymes) and 2 hr incubation under duodenal conditions (pH 6.8, + 37°C and duodenal enzymes). These were studied together and separately, as well as under pure acidic conditions without gastric enzymes. The yeast extracts contained equal amounts of allergen and were analysed by IgE‐immunoblotting. The acidic conditions had partly an enhancing and slightly degrading effect on the yeast allergens, whereas the gastric enzymes destroyed several allergens, including the important intermediate allergens of 31 and 45 kD. After treatment under both gastric and duodenal conditions most of the yeast allergens were destroyed, except mannan and a 10 kD protein component. The findings suggest that the allergen exposure caused by baker's yeast lakes place mainly on the mucosal surfaces orally and oesophageally and through viable baker's yeast organisms that manage to pass the stomach and duodenum and possibly lead to intestinal growth of the organism. Patients with IgE production against the 10 kD allergen and mannan are, however, moderately exposed to allergens consisting of soluble antigenic material only.
Venom‐specific IgG antibodies in bee and wasp allergy: lack of correlation with protection from stings
Authors: EWAN P. W. , DEIGHTON J. , WILSON A. B. , LACHMANN P. J.
Published: April 2006

This paper investigates the relationship between venom IgG levels and protection from stings. Venom‐specific IgG antibody levels have been measured by radioimmunoassay in untreated wasp‐ (n= 38) and bee‐allergic (n=16) patients presenting with systemic reactions to stings and in a sub‐group of these (wasp= 15; bee = 9), before and after the initial course of venom immunotherapy (VIT). A history was taken of all reactions, the last systemic reaction being graded on a scale of 1–8 and of the number and timing of stings. In untreated patients venom IgG levels were much higher in bee‐allergic patients (mean ± s.e. = 68.2 ± 7.1 % positive pool) than in the wasp group (27.1 ± 4.2%) (P< 0.05 Mann‐Whitney U‐test). There was a marked rise in venom IgG after the initial course of VIT in the wasp group (geometric mean and 95% confidence intervals = 40.5%, 28.8−54.3) but a much smaller rise in the bee group (15.3%, 6.6–24.1), with no overlap in the 95% confidence intervals. Bee patients, who were mainly beekeepers or their relatives, had been more heavily immunized with venom than wasp patients. They had received: (i) more stings (mean number of stings: bee, 26; wasp, 4; P< 0.001) and (ii) more stings per year. Wasp patients received their smaller number of stings over a much longer period, up to 40 yr. There was no correlation between the severity of the last systemic reaction and the venom IgG levels alone or venom IgG and IgE levels in combined analysis in either bee or wasp patients. This study shows that the pattern of IgG response differs in bee and wasp‐allergic subjects, and that most bee‐allergic subjects with systemic reactions have high levels of venom IgG. The degree of immunization with venom seems to be an important determinant of the venom IgG level. Our findings suggest thai venom‐specific IgG levels do not predict systemic reactions to stings and are not useful for monitoring VIT. If protection from stings is IgG‐mediated, our observations suggest that the relevant immune response is more complex, possibly involving IgG sub‐classes, IgG antibodies to individual venom antigens or antibody affinity, and not adequately reflected by measurement of the concentration of venom‐specific IgG.
The determinants of airway hyperresponsiveness to hypertonic saline in atopic asthma in vivo. Relationship with sub‐populations of peripheral blood leucocytes
Authors: SONT J. K. , BOOMS P. , BEL E. H. , BROUCKE J. P. VANDEN , STERK P. J.
Published: April 2006

In patients with asthma there is only a weak relationship between airway responsiveness to hypertonic saline and methacholine. We tested the hypothesis that airway responsiveness to hypertonic saline in asthma is related to the presence and activity of inflammatory cells in the peripheral blood. Nineteen atopic asthmatic adults (19–28 yr; PC20 0.06–12.4 mg/ml), not receiving steroid treatment, entered a methacholine and hypertonic saline period in random order. Dose‐response curves to doubling doses of inhaled methacholine (0.03–256 mg/ml) or hypertonic saline (0.9–14.4% NaCl) were obtained twice in each period, 7 days apart. The response was measured by FEV1 Methacholine responsiveness was measured by PC20 METH of FEV1 and responsiveness to hypertonic saline was expressed as the percentage fall in FEV, after 14.4% NaCl (HYP14.4%). Peripheral blood was collected before the second challenge test of each period. Apart from leucocyte counts and serum eosinophilic cationic protein (ECP) level, sub‐sets of lymphocytes (CD4 +/CD3 +, CD8 +/CD3 +, CD25 +/CD4 + and VLA‐1 +/CD4 +) were determined using flowcytornetry. HYP14.4% was positively correlated to basophil, eosinophil and monocyte counts (r = 0.64, 0.54 and 0.44, respectively; P< 0.05). The basophil count remained positively related to HYP 14.4% when PC20 METH or FEV1%pred were entered in multiple linear regression analyses (r = 0.66 and 0.75, respectively; P 0.05). There were no significant relationships between HYP14.4% or PC20 METH on one side and ECP level or T‐lymphocyte subsets on the other (P> 0.05). We conclude that airway responsiveness to hypertonic saline is positively related to the number of peripheral blood basophils, eosinophils and monocytes. Basophil count is an independent correlate of responsiveness to hypertonic saline, after correction for mcthacholine responsiveness and baseline lung function. This fits in with active involvement of basophils in airway narrowing to hypertonic saline in vivo.
Effect of murine recombinant interleukin‐5 on bronchial reactivity in guinea‐pigs
Authors: IWAMA T. , NAGAI H. , TSURUOKA N. , KODA A.
Published: April 2006

We have reported that an intratracheal injection of murine recombinant interleukin‐5 (mrIL‐5, 15 μg/0.0 ml/animal) induces the increased number of inflammatory leucocytes and epithelial cells in bronchoaiveolar lavage fluid (BALF) 24 hr after administration of mrIL‐5 in guinea‐pigs [1]. In this paper, we have examined the effects of mrIL‐5 on bronchial reactivity in guinea‐pigs. An intratracheal injection of mrIL‐5 (15 μg/0.0 ml/animal) induced airway hyperresponsiveness to acetytcholine (ACh) which was accompanied with eosinophilia and neutrophilia in blood at 24 hr in guinea‐pigs. Bovine serum albumin (BSA, 15 μg/0.0 ml/animal) used as a reference material did not cause airway hyperresponsiveness, blood eosinophilia and neutrophilia. Prednisotone (20 mg/kg, i.p.) inhibited mrIL‐5‐induccd airway hyperresponsiveness, eosinophilia and neutrophilia. Ketotifen (2 mg/kg, i.p.) also reduced this airway hyperresponsiveness and neutrophilia but not eosinophilia. In contrast, the injection of 2% disodium cromoglycale (DSCG) into the trachea showed the tendency of inhibitory effects against mrIL‐5‐induced airway hyperresponsiveness, eosinophilia and neutrophilia. The present data indicate that mrIL‐5 induces airway hyperresponsiveness, blood eosinophilia and neutrophilia in guinea‐pigs and that prednisolone and ketotifen inhibit mrIL‐5‐induced airway hyperresponsiveness accompanied with reduction of the increased number of leucocytes, suggesting that eosinophils and neutrophilia in blood may be important for the onset of bronchial hyperresponsiveness caused by mrIL‐5 in guinea‐pigs.
Effect of azelastine on substance P content in bronchoalveolar and nasal lavage fluids of patients with allergic asthma
Authors: NIEBER K. , BAUMGARTEN C. , RATHSACK R. , FURKERT J. , LAAKE E. , MÜLLER S. , KUNKEL G.
Published: April 2006

The study was carried out to investigate the effect of azelastine on the Substance P (SP) concentration in bronchoalveolar (BAL) and nasal (NAL) lavage obtained from atopic grass pollen asthmatics and non‐atopic healthy subjects. In BAL and NAL fluids there was a significant elevation in the baseline concentration of SP between asthmatics and volunteers. Allergen provocation induced a rise of SP in BAL and NAL in asthmatics, but not in volunteers. Azelastine pre‐treatment resulted in a significant reduction of SP in baseline concentration of SP in BAL and NAL from asthmatics. An elevation of SP in BAL or NAL fluids after allergen provocation was not seen in asthmatics pretreated with azelastine. Azelastine did not influence the SP concentration in BAL and NAL of volunteers.
Treatment of mosquito bites with cetirizine
Authors: REUNALA T. , BRUMMER‐KORVENKONTIO H. , KARPPINEN A. , COULIE P. , PALOSUO T.
Published: April 2006

Eighteen adult subjects sensitive to mosquito bites participated in a double‐blind, placebo‐controlled study with 10 mg cetirizine. The drug was given prophylactically and the subjects were then exposed to bites of Aedes communis mosquitoes in the field. Bite lesions were measured and pruritus was scored with a visual analogue scale at 15 min, 60 min, 12 hr and 24 hr. Cetirizine significantly decreased immediate wealing and pruritus and, interestingly, also had a clear effect on the delayed 12 hr and 24 hr bite papules and pruritus. The diameter of a 15 min mosquito‐bite weal was 10.10 ± 10.10 mm (mean ±s.d.) with the placebo and 5.5 ± 5.5 mm with cetirizine treatment (P<0.0). The 15 min pruritus scores were 36.36 ± 25.25 and 11.11±13.13 (P< 0.0), respectively. The diameter of the 24 hr mosquito‐bite lesion was 12.12 ± 21.21 mm with the placebo and 7.7±16.16 mm with cetirizine treatment (P<0.0). The 24 hr pruritus scores were 18.18 ± 25.25 and 6.6±14.14 (P<0.0), respectively. These results indicate that, in mosquito‐sensitive subjects, prophylactically administered cetirizine is an effective drug against both immediate and delayed mosquito‐bite symptoms.
Inhibition of neutral endopeptidase potentiates bronchoconstriction induced by neurokinin A in asthmatic patients
Authors: CRIMI N. , PALERMO F. , OLIVERI R. , POLOSA R. , MAORÌ S. , MISTRETTA A.
Published: April 2006

The endogenous tachykinins exhibit a range of properties which may he relevant in the pathophysiology of asthma. Their effects on the airways seem to be modulated by a variety of lung peptidases, including neutral endopeptidase (NEP). In order to evaluate the potential role of endogenous NEP activity in modulating tachykinins‐induced bronchconstriction in man in vivo, six atopic asthmatic patients, with a mean FEV1 value of 3.38 ± 0.76 1, and a histamine PD20 mean value of 0.024 mg. were studied. The influence of inhaled phosphoramidon (a potent NEP inhibitor) was examined against the NKA‐induced bronchospasm in a double‐blind, placebo‐controlled randomized study. Changes in airway calibre were followed as FEV1 and agonists responsiveness expressed as PD20 and PD15 for histamine and NKA respectively. Patients received nebulized phospharamidon sodium salt (10−5 M) or a control solution 10 min prior to the bronchoprovocation test with NKA. No significant difference was noticed between any of the study days and after inhaled phosphoramidon on baseline FEV1 values (3.29 ± 0.90 1) in comparison with the control solution (3.31 ± 0.79 1). Inhaled NKA produced a dose‐dependent fall in FEV1 values in all the subjects studied with a mean PD15 value of 20.91 × 10−9 mol. Phosphoramidon administered by inhalation elicited a significant (P < 0.0l vs baseline and control solution) potentiation in the airway responsiveness to inhaled NKA, the NKA PD15 value decreasing to 9.45 × 10−9 mol. The present study confirms that inhaled NKA induces a dose‐related bronchconstriction in asthmatic patients and demonstrates that inhaled phosphoramidon potentiates NKA‐induced bronchoconstriction.
Cloning and sequencing of the Dermatophagoides pteronyssinus group III allergen, Der p III
Authors: SMITH W.‐A. , CHUA K.‐Y. , KUO M.C. , ROGERS B. L. , THOMAS W. R.
Published: April 2006

House dust mites are widely recognized as major factors involved in the triggering of allergic diseases such as asthma. It is now apparent that the group III allergens of the Dermatophagoides mite species may play a significant role in a number of house dust mite allergic cases. Natural Der p III was isolated by gel filtration of salt precipitated Dermatophagoides pteronyssinus extract and as reported previously ran as a doublet of Mr 28 and 30 K on sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Natural Der fIII was isolated by affinity purification with the 5A12 monoclonal antibody. Amino acid sequence data was generated for both these proteins which was used to construct DNA probes to screen a Dermatophagoides pteronyssinus cDNA library by hybridization and resulted in the isolation of a recombinant Der p III cDNA clone, P3WSI. The 1059 bp cDNA fragment included a 786 bp open reading frame which encodes a pre‐pro region of 29 amino acids and a mature protein of 232 amino acids with a calculated Mr 24985. A search of the BLAST protein database has confirmed that the Der p III P3WS1 clone is approximately 50% homologous with other trypsin proteins. We have confirmed with both our natural protein sequence and the P3WS1 amino acid sequence data that the group III allergens are trypsin‐like proteins.
Epitope analysis of isoforms of the major allergen Phl p V by fingerprinting and microsequencing
Authors: PETERSEN A. , BECKER W.‐M. , SCHLAAK M.
Published: April 2006

The major allergen of timothy grass pollen (Phleum pratense), designated as Phl p V, consists of isoallergenic components of 38 and 32 kDa with pi values of 5.2 7.5 and 4.8 5 9, respectively. The different‐sized proteins reveal similarities in IgE reactivity, N‐terminal sequence and protein staining. For epitope analysis of these allergens u combination of enzymatic cleavage of electrophoretically separated proteins and iminunoblotting techniques with subsequent N‐terminal sequencing was performed. After isolation of the components from two‐dimensional PAGE gels. proteins were enzymatically cleaved and separated by SDS‐PAGE. By endoproteinase Glu‐C cleavage six IgE‐reactive fragments of each 32 kDa protein and three of each 38 kDa allergen were obtained. Microsequencing of the fragments revealed internal sequences that did not show any similarities between the different‐sized allergens. Therefore, we assume only slight structural variations among allergens of similar sizes, whereas the 32 and 38 kDa proteins reveal great differences.
Onset and duration of inhibition of ipratropium bromide nasal spray on methacholine‐induced nasal secretions
Authors: WAGENMANN M , BAROODY F. M. , JANKOWSKI R. , NADAL J. C. , ROECKER‐COOPER M. , WOOD C. C. , NACLERIO R. M.
Published: April 2006

We performed a randomized, double‐blind, placebo‐controlled cross‐over study with two different concentrations of ipratropium bromide (Atrovent®) nasal spray to evaluate its onset and duration of inhibition. Twenty‐four subjects with perennial rhinitis participated in the trial. Fifteen minutes to 12 hours after administration of ipratropium bromide (42 or 168 μg/nostril) or placebo nasal spray, methacholine challenges were performed and nasal secretion weights measured. After placebo administration the effect of methacholine remained unchanged over the 12‐h‐period. Both the 42 and 168 μg/nostril doses significantly inhibited the nasal hypersecretions induced by methacholine challenge within 15 min of treatment (P < 0.05). The 168 μg dose of ipratropium bromide continued to significantly reduce secretion weights through 6 hours, but the effectiveness of the 42 μg dose disappeared within 3 h. In addition to having a longer duration, the 168 μg/nostril dose produced approximately twice the inhibitory effect of the 42 μg dose.
Airways of allergic rhinitics are ‘primed’ to repeated allergen inhalation challenge
Authors: KOH Y. Y. , LIM H. S. , MIN K. U. , MIN Y. G.
Published: April 2006

The hypothesis that repeated exposure to a specific allergen will further increase bronchial responsiveness to that allergen is supported by indirect evidence. However, it has not been tested as intensely in the laboratory setting, and in some cases, conflicting results are presented. In order to test the hypothesis in the atopic subjects, allergen inhalation challenge tests were performed in 29 house dust mite (Dermatophagoides pternyssinus) sensitive subjects with allergic rhinitis. Nine subjects displayed early asthmatic responses (EARs) to the first challenge (Group 1). Twenty subjects with no significant airway response were submitted to the second challenge 24 h later. Thirteen subjects showed EARs (Group FI) and two of these showed late asthmatic responses (LARs)aswell. In Group II, there were significant changes between the first and second challenge in post‐allergen early phase FEV1, (88.1 ± 4.2 vs 71.7 ± 4.2%, baseline, P < 0.05) and in post‐allergen late phase FEV1, (93.1 ± 3.4 vs 86.6 ± 7.8. P < 0.05). After the second challenge. PD20 (provocative dose of methacholine required to produce a 20% fall in FEV1) decreased significantly from the baseline values. When challenged separately with twofold dose of allergen, only three and one of the Group II showed EAR and LAR respectively. PD20 did not change significantly after this challenge. These results indicated that two repeated exposure to allergen dose, which is not enough to cause significant airway responses at a time, may provoke asthmatic airway responses in the subjects with allergic rhinitis and that this effect of priming is not attributed to the cumulative dose but to the consequent effect of repeated allergen exposure.
Sputum eosinophilia after asthmatic responses induced by isocyanates in sensitized subjects
Authors: MAESTRELLI P. , CALCAGNI P. G. , SAETTA M. , DI STEFANO A. , HOSSELET J.‐J. , SANTONASTASO A. , FABBRI L. M. , MAPP C. E.
Published: April 2006

To assess the nature and the time–course of the cellular component of airway inflammation induced by isocyanates, we examined nine subjects with occupational asthma induced by toluene– or methylene diphenyl–diisocyanate (TDI, MDI) and four control subjects never exposed to isocyanates. Sputum was induced by inhalation of ultrasonically nebulized hypertonic saline (3–4% NaCl) before and 8, 24, 48 h after inhalation challenge with TDI or MDI. Expectorated samples were incubated with dithiothreitol, washed and cytocentrifuged. Differential cell counts were obtained on slides stained with May–Grünwald–Giemsa. Metachromatic cells (mast cells and basophils) were counted on slides stained with toluidine blue at pH 0.1. One occupational asthmatic exhibited a dual reaction to TDI, two exhibited a single early asthmatic reaction to M DI, six exhibited a late asthmatic reaction to TDI (n= 5) or M DI (n= 1), whereas no reactions were observed in control subjects after TDI challenge. In sensitized subjects eosinophils increased from a median value (interquartile range) of 5 (15)% before challenge to 29 (29)% (P= 0.014) and to 30 (31)% (P= 0.031) 8 and 24 h after TDI/MDI challenges, respectively. Sputum eosinophilia was observed both in early and late reactors and declined to near to baseline values 48 hr after challenge. Percentages of eosinophils in control subjects did not exceed 7% during the study. There was a significant correlation between the increase in eosinophil counts and the magnitude of the bronchoconstriction expressed as the area of FEV1 response to isocyanate challenge (rank correlation coefficient = 0–71, P= 0.014). No significant changes in sputum neutrophils, macrophages, lymphocytes and mast cells were detected. The results indicate that isocyanate–induced asthmatic responses are associated with influx of eosinophils in airway secretion lasting up to 24 h.
Role of eicosanoids in airflow obstruction and airway plasma exudation induced by trimellitic anhydride‐conjugate in guinea‐pigs 3 and 8 weeks after sensitization
Authors: ARAKAWA H. , LÖTVALL J. , LINDÉN A. , KAWIKOVA I. , LÖFDAHL C.‐G. , SKOOGH B.‐E.
Published: April 2006

Abstract. Trimellitic anhydride (TMA) is a low molecular weight chemical which can cause occupational asthma. We studied the role of eicosanoids in airway responses to TMA at different times after sensitization in actively sensitized guinea‐pigs. Sensitization was performed by two intradermal injections of free TMA (0.1 ml of 0.3% TMA in corn oil). At 3 and 8 weeks after sensitization, the guinea‐pigs were anaesthetized and challenged with intratracheal instillation of 0.5% TMA conjugated to guinea‐pig serum albumin (TM A‐GPSA; 50 μl). Lung resistance (RL) was measured to assess airflow obstruction, and the tissue content of Evans Blue dye was measured to assess airway plasma exudation. Intratracheal instillation of TMA‐GPSA induced a slowly progressing increase in RL, reaching a peak at approximately 3.5 min after the challenge (6.0 ± 2.0cm H2O/ml/s in the 3‐week group and 3.8 + 0.6 in the 8‐week group). Pretreatment before challenge with pyrilamine (anti‐histamine: 2 mg/kg. intravenously) slowed the onset of the increase in RL following challenge with TMA‐GPSA, and significantly attenuated the peak response. A combination of pyrilamine and IC1‐192, 605 (thromboxane receptor antagonist; 0.5 mg/kg, intravenously) completely abolished the increase in RLin both week groups. A combination of pyrilamine and ICI‐198, 615 (leukotrieneC4/D4/ E4 receptor antagonist: 0.5 mg/kg, intravenously) did not further attenuate the increase in RL compared with pretreatment with pyrilamine alone, but the induced Evans Blue dye extravasation was completely inhibited in the 3‐week group, whereas a remaining extravasation was observed in the 8‐week group. We conclude that the bronchoconstrictor response to TMA‐GPSA in actively sensitized guinea‐pigs is mediated by histamine and thromboxane A2 both early and late after sensitization, whereas leukotrienes and histamine partially mediate TMA‐induced airway plasma exudation.
Allergy to storage mites
Authors: TEE R. D.
Published: April 2006

Summary. There is now much evidence of sensitization to storage mites in urban populations as well as in the well‐documented rural populations. Sensitization is therefore not restricted only to those with occupational exposure.

There appears to be a limited allergenic crossreactivity between storage mites and house dust mites, although both species also possess their own unique allergens.

More research on identification and characterization of storage mite allergens and their crossreactivity is needed to understand their complexity. Such studies are necessary to obtain high quality extracts for diagnosis and possible immunotherapy.

Development of immunoassays employing MoAbs will allow measurement of storage mite concentrations in workplaces and houses and the study of exposure‐response relationships.
cDNA sequence analysis of the main olive allergen, Ole e I
Authors: LOMBARDERO M. , BARBAS J. A. , DEL PRADO J. MOSCOSO , CARREIRA J.
Published: April 2006

Olea europaea (Ole e) I‐specific cDNA sequences were amplified by 3′‐RACE‐PCR, using specific primers based on the N‐Terminal sequence of the allergen, and cloned into appropriate vectors. The nucleotide sequence data obtained revealed the presence of isogenic variation in Ole e I gene(s). The molecular mass, pI, amino acid composition and sequence of the predicted polypeptides agree with data previously obtained by analysis of purified Ole e I from pollen. Furthermore, by treatment of purified Ole e I with specific glycopeptide hydrotases it has been demonstrated the presence of N‐glycosylation in the allergen, and there is a unique concensus site for N‐Linked glycosylation at positions 111‐113 of the deduced amino acid sequence. The Ole e 1 predicted sequence shows a significant homology with three putative proteins encoded respectively by the anther‐specific LAT52 gene from tomato and the pollen specific genes Zmcl3 from maize and OSPSG from rice, suggesting that these proteins could have a role in one of the development processes unique to male gametophytes.
Familial atopy in Australian pedigrees: adventitious linkage to chromosome 8 is not confirmed nor is there evidence of linkage to the high affinity IgE receptor
Authors: BRERETON H. M. , RUFFIN R. E. , THOMPSON P. J. , TURNER D. R.
Published: April 2006

Atopy frequently displays autosomal dominant inheritance and recent studies have favoured genetic linkage between atopy and the human chromosome 11q13. We have studied 12 extended families with aggregation of atopy consistent with autosomal dominant inheritance. The families have been studied for linkage of asthma and atopy to loci on chromosome 8p following the observation that one family suggested preliminary evidence of linkage to an anonymous hypervariable locus cloned from a DNA fingerprint and mapped to 8pter‐p22. Subsequent analysis shows this putative linkage to be adventitious as the remaining 11 families do not support linkage between atopy and 8p, We have analysed the same families for evidence of linkage of atopy to loci on 11q13. In these families there is no evidence of association between atopy and the 11q loci stronger than that expected by chance alone; furthermore there is no suggestion subpopulation of these families display linkage between atopy and the loci. In addition neither the 8p loci nor the 11q loci exhibit evidence of linkage to atopy by affected sib‐pair analysis. This also conflicts with previously published data for 11q.
Dermatophagoides pteronyssinus II‐induced interleukin‐4 and interferon‐γ expression by freshly isolated lymphocytes of atopic individuals
Authors: BYRON K. A. , O'BRIEN R. M. , VARIGOS G. A. , WOOTTON A. M.
Published: April 2006

Cytokines are known to play a major role in mediating many of the immunological and pathological features of allergic disease. Much of our understanding of cytokine production in response to allergens has come from studying allergen‐specific T cell clones following long‐term in vitro culture. This has largely been due to the lack of sufficiently sensitive assays to measure allergen‐induced cytokine production by freshly isolated peripheral blood mononuclear cells (PBMCs). Here we have used the polymerase chain reaction to amplify reverse transcribed interleukin‐4 (IL‐4) and IFNγ mRNA expressed by allergen‐stimulated PBMCs from a variety atopic individuals. Using Der p II, a major allergen of the house dust mite (HDM) Dermatophagoides pteronyssinus, we have demonstrated that cells from HDM‐sensitive atopic patients (n= 12), can be induced to express either IL‐4 alone (three patients), IL‐4 and IFNγ (six patients), IFNγ alone (two patients) or neither cytokine (one patient). Cells from 13 non‐atopic control individuals were also stimulated with Der p II and cytokine mRNA production was studied. None expressed IL‐4, while seven of 13 transcribed IFNγ. Our results suggest that atopic individuals have allergen‐reactive T cells at various stages of differentiation, with respect to the cytokines they produce. The use of this technique will aid in the further understanding of specific cellular hypersensitivity in allergic disease.