bluuebunny/crossref_metadata_2025 huggingface dataset:
The 40‐kilodalton allergen of Candida albicans is an alcohol dehydrogenase: molecular cloning and immunological analysis using monoclonal antibodies
Authors: SHEN HORNG‐DER , CHOO KONG‐BUNG , LEE HSIEN‐HSIUNG , HSIEH JIA‐CHING , LIN WIN‐LING , LEE WEN‐RONG , HAN SHOU‐HWA
Published: April 2006

To characterize the 40‐kilodalton (kD) major allergen of Candida albicans (C. albicans), six monoclonal antibodies (MoAbs) against this allergen were generated. In SDS‐polyacrylamide gel electrophoresis and immunoblot analysis, these MoAbs showed four different reaction patterns to antigens of six different Candida species. With the exception of one MoAb, other MoAbs were resistant to periodate treatment indicating non‐carbohydrate epitopes were probably being recognized by these MoAbs. These MoAbs were used in the molecular cloning and immunological analysis of the gene coding for the 40‐kD allergen. Nucleotide sequence determination of the two λgtl 1 cDNA clones obtained showed that the 40‐kD allergen is an alcohol dehydrogenase (ADH) which shares a 70% amino acid sequence homology with the ADH isozyme I of Saccharomyces cerevisiae. This finding was confirmed by positive immunological response of the lysates of the clones obtained and a preparation of ADH of Saccharomyces cerevisiae to various MoAbs and to IgE antibodies in sera of allergic patients.
Cord serum IgE: an insensitive method for prediction of atopy
Authors: HIDE D. W. , ARSHAD S. H. , TWISELTON R. , STEVENS M.
Published: April 2006

Cord total serum IgE has been advocated as a screening test to detect infants at high risk of allergy who would be suitable for preventive measures. In a population based prospective study to look at the predictive capacity of cord IgF and family history of atopy 1111 one‐year‐old infants were followed‐up. Cord IgE was measured using the EIA ultra technique (Pharmacia, Uppsala, Sweden). Atopic symptoms developed in 255 (23%) at one year, 183 (16·5%) had probable atopy (clinical disorder but negative skin prick test (SPT)) and 72 (6·5%) had definite atopy (clinical disorder with positive SPT), There was no difference in the mean cord IgE levels in infants with or without atopic manifestations. The cut‐off for IgF was taken at 0·6 ku/1. The specificity of the test was 92% but the sensitivity was only 8·5%. The positive and negative predictive values were 24% and 78% respectively. Family history of atopy is far more sensitive in detecting infants at risk of atopy and little is added by knowledge of cord IgE.
TNFα mRNA expression in allergic inflammation
Authors: YING SUN , ROBINSON D. S. , VARNEY V. , MENG QIU , TSICOPOULOS A. , MOQBEL R. , DURHAM S. R. , KAY A. B. , HAMID Q.
Published: April 2006

Using the technique of in situ hybridization, we have attempted to identify messenger RNA for tumour necrosis factor‐alpha (TNFα) in cells infiltrating allergen‐induced late phase reaetion (LPR) of the skin and the nose ofatopic subjects. We have also compared the number of TNFα mRNA positive cells in bronchoalveolar lavage (BAL) from atopic asthmatics and normal controls. Twenty‐four hours after local allergen challenge, 12/14 skin biopsies and 9/10 nasal biopsies had positive hybridization signals for TNFα mRNA whereas only 4/14 and 2/10 biopsies were positive in the relevant diluent controls. Compared with diluent sites significantly increased numbers of cells expressing mRNA for TNFα were observed in the LPR of skin (P <0.004) and nose(P <0.006). All BAL from asthmatics (n= 10) and from normal volunteers (n= 10) had cells showing positive hybridization signals for TNFα mRN A but these were at increased frequency in asthmatics (P< 0.001). These results suggest that TNFα may be an important cytokine in atopic allergic inflammation.
The IgG subclass antibody response to an inhalant antigen (Dermatophagoides pteronyssinus) during the first year of life: evidence for early stimulation of the immune system following natural exposure
Authors: MARIANI F. , PRICE J. F. , KEMENY D. M.
Published: April 2006

This study investigates the early humoral immune response following natural exposure to an inhalant antigen (Dermatophagoides pteronyssinus) in 36 babies, from birth until 1 year of age. The total IgG and subclass 1 and 4 D. pteronyssinus‐specific antibody levels were assayed in sera collected at 7 days, 3 and 12 months by ELISA. After an initial fall, due to the progressive loss of maternal antibodies, an IgG specific response to D. pteronyssinus was seen between 3 and 12 months. This was restricted to the IgG1 subclass when the values at 12 months were significantly higher than those detected at the third month (P < 0.001, paired t test). No D. pteronyssinus‐specific IgG4 antibody was detected in any subject at any of the time points tested. The present study demonstrates that inhalant as well as food antigens are able to stimulate the immune system during the first year of life and that the antibodies produced are of the IgG1 subclass.
Effects of single and multiple inhalations of platelet‐activating factor on airway cell composition and responsiveness in monkeys
Authors: WEGNER C. D. , CLARKE C. C. , TORCELLINI C. A. , LETTS L. G. , GUNDEL R. H.
Published: April 2006

Platelet‐activating factor (PAF) is a potent pro‐inflammatory mediator that may play a role in the pathogenesis of airway hyper‐responsiveness and asthma. In man, a single inhalation of PAF induces a small but prolonged increase in airway responsiveness in some individuals. The purpose of this study was to determine the effects of single and multiple inhalations of PAF on airway cell composition and responsiveness in monkeys. Anaesthetized and intubated adult male cynomolgus monkeys were studied. Airway cell composition was measured by bronchoalveolar lavage (BAL). Airway responsiveness was measured by determining the concentration (PC100) of inhaled methacholine that caused a 100% increase in respiratory system resistance (Rrs). Airway cell composition (BAL) and responsiveness (PC100) were determined 1 day before and 20 hr after a single inhalation of PAF (∼200 μg) or 3 days before (Day 0) and 3 days after (Day 10) 3‐alternate‐day (Days 3, 5 and 7) inhalations of PAF (each ∼600 μg). The single inhalation of PAF (n= 8) caused an acute increase in Rrs (147±69%), an increase in BAL granulocytes, and a decrease in PC100 in four of eight animals that was moderate (>eight fold) in only one animal. The mean ±s.e. change in log PC100 was −0.29±0.18. The multiple inhalations of PAF (n=8) caused acute increases in Rrs (143±38%, 175±44% and 156±39%, respectively), an increase in BAL granulocytes, and a decrease in PC100 in four of eight animals that was moderate in two animals. The mean±s.e. change in log PC100 was −0.43±0.22. In summary, single and multiple inhalations of PAF in monkeys induce a granulocyte airway infiltration and, in some animals, an increase (usually small) in airway responsiveness.
Lack of linkage between atopy and locus 11q13
Authors: HIZAWA N. , YAMAGUCHI E. , OHE M. , ITOH A. , FURUYA K. , OHNUMA N. , KAWAKAMI Y.
Published: April 2006

Atopy as defined in terms of IgE responsiveness was reported to be controlled by a single gene in British families, and this concept was further supported by a significant linkage between atopy and restriction fragment length polymorphism (RFLP) detected by a DNA probe specific to chromosome 11q13. To confirm this observation in a Japanese population, segregation and linkage analyses were done in four large families. Although segregation patterns of atopy were in agreement with the pattern of autosomal dominant inheritance, there was no significant linkage between atopy and locus 11q13. Alterations in the definitions of atopy did not affect the results. These findings suggested the presence of heterogeneity in genetic elements of atopy, even though atopy may be determined mainly by a single dominant gene.
Atopy and bronchial hyperresponsiveness: exclusion of linkage to markers on chromosomes 11q and 6p
Authors: AMELUNG P. J. , PANHUYSEN C. I. M. , POSTMA D. S. , LEVITT R. C. , KOETER G. H. , FRANCOMANO C. A. , BLEECKER E. R. , MEYERS D. A.
Published: April 2006

Previous studies have reported a familial predisposition for the development of atopy, bronchial hyperresponsiveness and clinical asthma, and therefore have suggested the presence of a heritable component to these disorders. The specific contributions of genetic and environmental factors in the pathogenesis of allergic disease and asthma have not been determined although Cookson et al. [1] have postulated linkage between atopy and chromosome 11q. We have studied 20 families (two and three generations) ascertained through a proband identified as having asthma (90% were also allergic) during the period of time between 1962 and 1970. Of those who were originally skin test positive, 82% remained positive. All probands whose pulmonary function allowed retesting (FEV1 > 1.21) remained hyperresponsive to histamine. The children of these probands are now in the same age range as their parents when they were originally evaluated; 66% are atopic using criteria described by Cookson et al. (one or more positive skin tests ≥2 mm, an elevated total serum IgE or a positive specific IgE) and 22% demonstrate bronchial hyperresponsiveness (PC20 FEV1) to histamine. Using the highly polymorphic marker INT2 (which maps 2 cM from pλMS.51 on chromosome 11q) and atopy, we obtained a lod score of – 2.00 at a recombination fraction of 0.12. In addition, because many studies have suggested an association between atopy and certain HLA antigens, we investigated the possibility of linkage between atopy and bronchial hyperresponsiveness and D6S105, a polymorphic marker on chromosome 6p, located 7 cM from HLA‐DR. For this marker and atopy, we observed a lod score of – 2.00 with a recombination fraction of 0.07. Similar results were observed with both of these markers and bronchial hyperresponsiveness. By restudying these probands as well as their family members, we were unable to find evidence for linkage between atopy or bronchial hyperresponsiveness and these regions of chromosomes 11 and 6 in this population.
Inhibition of proliferative responses of lymphocytes to food antigens by an anti‐allergic drug, N(3′,4′‐dimethoxycinnamoyl) anthranilic acid (Tranilast) in children with atopic dermatitis
Authors: KONDO N. , FUKUTOMI O. , KAMEYAMA T. , ORII T.
Published: April 2006

Experimental studies have shown that N(3′, 4′‐dimethoxycinnamoyl) anthranilic acid (Tranilast) inhibits reaginic antibody‐mediated hypersensitivity reactions, and it has been demonstrated to be an effective drug for patients with bronchial asthma. On the other hand, from the nature of the cellular infiltrate seen in eczematous lesions, it appears that some form of cell‐mediated immunity may be involved in addition to IgE‐mediated immunity in the pathogenesis of atopic dermatitis (AD). Moreover, we have previously reported that the proliferative responses of peripheral blood mononuclear cells (PBMCs) to ovalbumin (OA) or bovine serum albumin (BSA) in children with AD who are sensitive to hen's egg or cow's milk were significantly higher than those of healthy children and hen's egg or cow's milk sensitive children with immediate symptoms.

In this study, we have showed that the proliferative responses of PBMCs to OA were dose‐dependently inhibited by Tranilast on patients with AD. The responding cells to OA were shown, through separation experiments, to be T cells, and the proliferative responses of T cells to OA were also dose‐dependently inhibited by Tranilast. Moreover, the inhibition was thought to occur at the initial stage of the proliferative reactions. These results suggest that Tranilast can be clinically applied to patients with AD.
Purification and characterization of Lep d I, a major allergen from the mite Lepidoglyphus destructor
Authors: VENTAS P. , CARREIRA J. , POLO F.
Published: April 2006

A major allergen of the storage mite Lepidoglyphus destructor (Lep d I) has been purified by affinity chromatography using an anti‐Lep d I monoclonal antibody. The purity of the protein obtained by this procedure was assessed by reverse‐phase HPLC. Lep d I displayed a molecular weight of 14 kD on SDS‐PAGE under non‐reducing conditions, and 16 kD in the presence of a reducing agent. Analytical IEF revealed a little charge microheterogeneity, showing three bands with pIs 7.6–7.8. Purified Lep d I retained IgE‐binding ability, as proved by immunoblotting experiments after SDS‐PAGE and RAST with individual sera from L. destructor‐sensitive patients. Results from the latter technique demonstrated that 87% of L. destructor‐allergic patients had specific IgE to Lep d I, and a good correlation between IgE reactivity with L. destructor extract and Lep d I was found. In addition, RAST inhibition experiments showed that IgE‐binding sites on Lep d I are major L. destructor‐allergenic determinants, since Lep d I could inhibit up to 75% the binding of specific IgE to L. destructor extract; on the other hand. Lep d I did not cross‐react with D. pteronyssinus allergens.
Group V allergens in grass pollens: IV. Similarities in amino acid compositions and NH2‐terminal sequences of the Group V allergens from Lolium perenne, Poa pratensis and Dactylis glomerata
Authors: KLYSNER S. , WELINDER K. G. , LØWENSTEIN H. , MATTHIESEN F.
Published: April 2006

Monoclonal antibodies (PpV4) raised against Phleum pratense group V allergen were used for immuno‐affinity chromatography of cross‐reacting group V allergens from related grass species. Fractions enriched in group V allergen were obtained from Lolium perenne, Poa pratense and Dactylis glomerata extracts. The major components in these fractions were found in the Mwr range 25–28 kD. IgE binding to these components was shown using a pool of grass allergic sera, by SDS‐PAGE immunoblotting. These fractions were electroblotted from tricine SDS‐PAGE gels onto a polyvinylidenedifluoride membrane and selected group V bands were directly cut out and used for amino acid analysis and NH2‐terminal sequencing.

Both the amino acid compositions and the NH2‐terminal sequences obtained for each group V allergen were almost similar to each other and to the sequence and composition of the previously described allergen Phl p V from Phleum pratense. A common trait of the investigated allergens, is the very high contents of alanine (25–32%) and the presence of the modified amino acid, hydroxyproline.
The role of intercellular adhesion molecule‐1 in chronic airway inflammation
Authors: GUNDEL R. H. , WEGNER C. D. , TORCELLINI C. A. , LETTS L. G.
Published: April 2006

We have examined the role of intercellular adhesion molecule‐1 (ICAM‐1) in chronic airway inflammation and airway hyperresponsiveness in a primate model of asthma. Airway cellular composition was assessed by bronchoalveolar lavage (BAL) and airway responsiveness was measured as the bronchoconstrictor response to inhaled methacholine. In animals with chronic airway inflammation (increased BAL eosinophils) and sustained airway hyperresponsiveness, a 7 day dosing scheme with a murine anti‐human ICAM‐1 monoclonal antibody (R6.5, 2 mg/kg/day; i.v.) did not reduce the existing airway inflammation or airway hyperresponsiveness. In contrast, a similar dosing scheme with dexamethasone (0.2 mg/kg/day, i.m.) was found to significantly reduce both the airway eosinophilia and hyperresponsiveness. However, one week after cessation of dexamethasone treatment, the airway inflammation and hyperresponsiveness returned to pre‐treatment levels. In further experiments where animals were first treated with dexamethasone (7 days) followed by a 7 day treatment with R6.5, the reoccurrence of airway inflammation and subsequent increase in airway responsiveness was prevented. We conclude that the efficacy of ICAM‐1 is primarily associated with inhibition of the influx of inflammatory cells into the airways and subsequent reduction in airway responsiveness. These data suggest that in lungs with pre‐existing inflammation the modulation of ICAM‐1 following treatment with glucocorticoids may be a novel and more selective long‐term treatment for control of the chronic airway inflammation and hyperresponsiveness associated with bronchial asthma.
Specific immunological and bronchopulmonary responses following intradermal sensitization to free trimellitic anhydride in guinea pigs
Authors: HAYES J. P. , DANIEL R. , TEE R. D. , BARNES P. J. , CHUNG K. F. , NEWMAN TAYLOR A. J.
Published: April 2006

We have developed a guinea pig model of trimellitic anhydride‐induced airway hypersensitivity responses. In one group of guinea pigs, injected intradermally with 0.1 ml 30% trimellitic anhydride (TMA), we examined the specificity of the bronchopulmonary response to TMA comparing the effect of intravenous TMA conjugated to guinea pig serum albumin (GPSA) with a control hapten (procion dye) protein conjugate (PD‐GPSA). A significant increase in pulmonary inflation pressure (PIP) was provoked in sensitized animals following intravenous injection with TMA‐GPSA (20%; 0–400, median; range) as compared to intravenous injection of PD‐GPSA. In the second group we compared three different methods of sensitization: single injection of 0.1 ml of 0.3% TMA; four injections of 0.1 ml of 0.1% TMA; and a single high dose injection of 30% TMA. Following intravenous TMA‐GPSA guinea pigs sensitized with a single injection 0.3% TMA had an increase in PIP of 395%; 220–600, while those given four repeat injections of 0.1% TMA had an increase in PIP of 343%; 315–490. These results were significantly higher than the increase in PIP (160%; 0–220) which occurred in guinea pigs sensitized with a single dose of 30% TMA. Four of 11 guinea pigs given low dose injections of TMA had bronchopulmonary responses to inhaled TMA‐GPSA. All sensitized guinea pigs had specific IgG1 antibodies demonstrated by enzyme linked immunosorbent assay (ELISA) and confirmed by ELISA inhibition. Four guinea pigs sensitized by low dose injections of TMA had IgE antibodies demonstrated by passive cutaneous anaphylaxis.

We conclude that intradermal sensitization to free TMA induces a specific immune and airway hypersensitivity response to TMA‐GPSA. Single low dose injection of TMA is the most effective method of sensitization. Intradermal sensitization to free TMA may be a valuable method of sensitization for the developing of an animal model of occupational asthma caused by low molecular weight chemicals.
Dynamics of mast cells in the nasal mucosa of patients with allergic rhinitis and non‐allergic controls: a biopsy study
Authors: FOKKENS W. J. , GODTHELP T. , HOLM A. F. , BLOM H. , MULDER P. G. H. , VROOM Th. M. , RIJNTJES E.
Published: April 2006

Mast cell degranulation is thought to be an important component of the pathogenesis of allergic rhinitis. Quantitative studies on mast cells in nasal mucosa after allergen exposure have given widely divergent results, ranging from an overall decrease via redistribution to an overall increase. We investigated this problem by employing a combination of anti‐IgE and toluidine blue staining of biopsy specimens. In allergic patients anti‐IgE was found to identify all mast cells and toluidine blue to detect mast cells that were not (totally) degranulated.

The study was composed of two parts done in different patient groups. In the first part of the study biopsies were performed in 23 patients with isolated grass‐pollen allergy, once during natural provocation in the summer and once in the winter. Biopsies were also performed in 12 controls. Non‐allergic controls were found to have the same number of mast cells in the lamina propria as asymptomatic allergic patients. The controls seldom have mast cells in the epithelium. The patients with isolated grass‐pollen allergy showed an increase in the numbers of mast cells in the lamina propria during natural provocation and the same seemed to occur in the epithelium as well. During natural provocation almost all of the mast cells in the epithelium and half of those in the lamina propria were degranulated.

In the second part of the study 17 patients with isolated grass‐pollen allergy and four controls were challenged daily with allergen extract during a 2‐week period in the winter. During this period biopsies were performed at eight different occasions, i.e. once before, six occasions during and once after the provocation period. The results of this part of the study showed that during provocation mast cells migrate to the surface of the nasal mucosa, where they become degranulated, and that the pool of mast cells in the lamina propria was apparently replenished by migration of mast cells from the vessels in the lamina propria. The total number of mast cells in the lamina propria remained approximately the same while the mast cells residing in an increasingly thick layer measured from the basal membrane into the lamina propria became degranulated. After 2 weeks, 82% of the mast cells in the lamina propria was degranulated and it was only in the deepest layers that some toluidine blue positive cells were found.

This study can explain the seemingly conflicting reports in the literature on mast cell dynamics and degranulation and shows that the reported differences are due to differences in the techniques used and the time of evaluation.
Investigation of the tendency to wheeze in pollen sensitive patients
Authors: ARMITAGE J. M. , LAM K. SIN FAI , WILKINSON I. , FAUX J. A. , HOPKIN J. M.
Published: April 2006

We have undertaken a double blind placebo controlled study of the effect of nasal beclomethasone on the tendency to wheeze in 20 unselected hay fever sufferers, half with a history of previous seasonal wheezing. We found no difference between either bronchial hyperresponsiveness, as measured by methacholine challenge, home‐monitored PEFR, nor recorded wheeze nor cough between treated and placebo groups although the numbers were small. All were allowed the antihistamine cetirizine hydrochloride 10 mg daily. Eighteen out of the 19 patients had either bronchial hyper‐responsiveness (PD20 methacholine < 8 μmol or a > 2 doubling dose change in their PD20 during the pollen season). We have shown a significant positive correlation between a hay fever score (HFS) (created by taking the sum of the home scored; nasal discharge, nasal blockage, eye irritation, sneeze and antihistamine use) and peak seasonal specific IgE to mixed grass pollen (Spearman correlation coefficient 0.5 P<0.02). There was also a positive correlation between the rise in specific IgE from pre to peak season and the HFS, correlation coefficient 0.6 P =0.03).
The allergens of dog II. Identification and partial purification of a major dander allergen
Authors: FORD A. W. , KEMENY D. M.
Published: April 2006

A dog hair and dander (DHD) extract was prepared from hair obtained from mixed breeds. By SDS‐polyacrylamide gel electrophoresis (SDS–PAGE) and immunoblotting, using sera from 32 dog‐allergic subjects, a number of IgE radio‐staining bands could be seen. In 78% of sera a protein of molecular weight (MW) of 21 000 daltons, designated Ag X, was found to bind IgE and in 34% it did so strongly. This allergen was isolated from DHD by size‐exclusion and ion exchange chromatography. The final product was a single allergen of MW of 21 000 and an isoelectric point of approximately 5.2. An additional protein‐staining band could still be seen of MW of 24 000 daltons. Using a serum which contained IgE antibodies only to Ag X, this allergen was found only in DHD extract and dog saliva and was absent from dog serum and urine. It was the same dog allergen that we [1] reported as Ag 8 using crossed radio‐immunoelectrophoresis (CRIE) and that Blands et al. [2] and Løwenstein [3] described as Ag 13. We propose that this major dog allergen be given the title Can f I according to the new allergen nomenclature.
Localization of allergens in the domestic mite Lepidoglyphus destructor
Authors: VAN HAGE‐HAMSTEN M. , SCHEYNIUS A. , HÄRFAST B. , WIRÉN A. , JOHANSSON S. G. O.
Published: April 2006

The non‐pyroglyphid domestic mite Lepidoglyphus destructor is a major source of allergen causing respiratory symptoms in farming environments. This study is the first to focus on the localization of the allergens in the non‐pyroglyphid domestic mite Lepidoglyphus destructor. Cryostat‐cut sections of L. destructor mite bodies and faecal pellets were probed with one of three mouse monoclonal antibodies (MoAbs) raised against L. destructor or with patient sera, and stained with immunoperoxidase. Eight sera were RAST‐positive to L. destructor and L. destructor faecal pellets. These eight RAST‐positive patient sera labelled the wall of the L. destructor gut and two of them also the faecal pellets. The MoAbs additionally labelled parts of the exoskeleton and reacted with a majority of the faecal pellets. Cryostat‐cut sections of Dermatophagoides pteronyssinus mite bodies and faecal pellets were probed with L. destructor MoAbs, which resulted in only slight staining of a few faecal pellets. The results suggest that at least one L. destructor allergen is associated with digestion.
Inhibitory effects of formoterol and terbutaline on the development of late phase skin reactions
Authors: GRÖNNEBERG R. , ZETTERSTRÖM O.
Published: April 2006

The capacity of the β2‐agonist terbutaline and the longer‐acting β2‐agonist formoterol to suppress the development of late phase skin reactions to anti‐human IgE was evaluated in 17 healthy volunteers. Anti‐IgE injected intradermally per se induced an early weal and flare reaction, followed by a progressively increasing induration, the LCR, with a duration of ≥ 24 hr. The LCR was inhibited by 40% when the weal was infiltrated with formoterol 250 ng 30 min after challenge (n=9, P < 0.01). The same anti‐LCR effect was achieved by compensating for the shorter duration of action of terbutaline with repeated drug infiltration in 12.5 μg doses of the weal produced by anti‐IgE up to 3½ hr after challenge (n=8). The data support the hypothesis that β2‐agonists, both short‐and long‐acting, inhibit IgE‐dependent LCRs by preferentially interacting with inflammatory events after the initial mast cell degranulation
Eosinophil cationic protein in peripheral blood of pediatric patients with allergic diseases
Authors: SUGAI T. , SAKIYAMA Y. , MATUMOTO S.
Published: April 2006

We have studied the levels of eosinophil cationic protein (ECP) and tumour necrosis factor (TNF) in peripheral blood obtained from 68 children with bronchial asthma and 11 children with atopic dermatitis. The ECP mean concentrations of the patients were 23.7 ± 21.4 μg/1 and 21.2 ± 18.7 μg/1 for bronchial asthma and atopic dermatitis respectively, which were significantly higher than the control value, 5.8 ± 2.3 μg/1 (P < 0.005). TNF was unmeasurable in almost all the samples and no significant difference was observed between normal controls and asthmatic children. A significant correlation was observed between serum levels of ECP and blood eosinophil counts in both diseases (r=0.873; P < 0.01 and r=0.740; P < 0.01, respectively). However, no obvious correlation was observed between serum levels of ECP and IgE levels. ECP levels were significantly reduced by treatment and normalized in parallel with blood eosinophil counts in the patients with total IgE levels less than 800 U/ml. Irrespective of the total IgE levels, the reduction of serum ECP levels was correlated with a decrease in the number of asthmatic attacks and/or improvement of pulmonary function. These results suggest that the ECP levels in peripheral blood indicate an increased activity of eosinophil and would be a more useful marker than eosinophil counts for making clinical analyses and estimating treatment efficacy in paediatric patients with allergic diseases.
Altered distribution of IgG subclasses in aspirin‐induced asthma: high IgG4, low IgG1
Authors: SZCZEKLIK A. , SCHMITZ‐SCHUMANN M. , NIZANKOWSKA E. , MILEWSKI M. , ROEHLIG F. , VIRCHOW CHR.
Published: April 2006

We have determined IgG subclass concentrations in 100 patients with aspirin‐induced asthma and 80 healthy controls. Patients on chronic corticotherapy (n= 64) had significantly lower total IgG levels than patients not receiving steroids (n= 36) or controls. Corticotherapy was not associated with changes in the subclass distributions. In patients, the most striking finding was elevation of IgG4. It was not related to corticotherapy or serum IgE levels. The rise in IgG4 was accompanied by a modest, though statistically significant, depression of IgG1. No changes of IgG2 and IgG3 concentrations were observed. Thus, aspirin‐induced asthma is characterized by a distinct pattern of distributions of IgG subclasses. It is suggested that in aspirin‐induced asthma elevation of IgG4 might result from chronic antigenic stimulation, of viral origin, and that determination of IgG subclass distribution might be of clinical interest.
The onset and recovery from airway hyperresponsiveness: relationship with inflammatory cell infiltrates and release of cytotoxic granule proteins
Authors: GUNDEL R. H. , WEGNER C. D. , LETTS L. G.
Published: April 2006

Previous studies from our laboratory have demonstrated a temporal relationship between eosinophil influx into the airways and the onset of airway hyperresponsiveness to inhaled methacholine. The purpose of the present study was to extend this observation by evaluating changes in airway cellular composition and measuring the levels of granulocyte‐derived mediators recovered in BAL fluid during the onset and recovery from antigen‐induced airway hyperresponsiveness. Airway cellular composition, airway responsiveness to inhaled methacholine and the levels of BAL fluid EPO and MPO were monitored over a 32 day study in eight adult male Ascaris suum sensitive cynomolgus monkeys. Repeated Ascaris suum inhalation (nine challenges during days 0–21) resulted in a selective, sustained airway eosinophilia that was temporally related with the onset and maintenance of airway hyperresponsiveness (r= 0.67, P < 0.001). The level of BAL eosinophil‐derived EPO was increased and remained elevated concurrent with the increase in airway eosinophils and airway responsiveness. During the recovery phase (days 22–32) the actual number of eosinophils remained elevated, while BAL EPO levels were significantly decreased. The recovery phase was also associated with a transient increase in the number of BAL neutrophils and MPO concentration. We conclude that the number and state of activation of airway eosinophils directly correlate with the onset and maintenance of airway hyperresponsiveness. Recovery from airway hyperresponsiveness is associated with a decrease in eosinophil activation and a transient increase in the number of activated neutrophils.
Viruses as precipitants of asthma symptoms. I. Epidemiology
Authors: PATTEMORE P. K. , JOHNSTON S. L. , BARDIN P. G.
Published: April 2006

Christopher Robin had wheezles and sneezles, they bundled him into his bed.

They gave him what goes with a cold in the nose, and some more for a cold in the head.

They wondered if wheezles could turn into measles, if measles would turn into mumps;

They examined his chest for a rash, and the rest of his body for swellings and lumps.

They sent for some doctors in sneezles and wheezles, to tell them what ought to be done.

All sorts and conditions of famous physicians came hurrying round at a run.

They all made a note of the state of his throat, they asked if he suffered from thirst;

They asked if the sneezles came after the wheezles, or if the first sneezle came first.

They said, ‘If you teazle a sneezle or wheezle, a measle may easily grow.

But humour or pleazle the wheezle or sneezle, the measle will certainly go.’

They expounded the reazles for sneezles and wheezles, the manner of measles when new.

They said ‘If he freezles in draughts and in breezles, then PHTHEEZLES may even ensue.’

Christopher Robin got up in the morning, the sneezles had vanished away.

And the look in his eye seemed to say to the sky, ‘Now, how to amuse them today?’

A. A. Milne, Now We Are Six, 1927.
Effect of corticosteroid treatment on interleukin‐1 and tumour necrosis factor secretion by monocytes from subjects with asthma
Authors: VECCHIARELLI A. , SIRACUSA A. , CENCI E. , PULITI M. , ABBRITTI G.
Published: April 2006

Peripheral blood monocytes (PBM) may be activated in asthmatic patients, a condition usually reverted by corticosteroid (CS) treatment. In the present research we have evaluated the spontaneous or lipopolysaccharide (LPS)‐induced production of interleukin 1 (IL‐1) and tumour necrosis factor (TNF) by PBM obtained from 14 asthmatic subjects during an asthmatic attack and after 1 week of CS treatment. The control group included 20 healthy volunteers. PBM obtained during severe asthma showed a pattern of IL‐1 and TNF secretion similar to that of normal subjects. After CS treatment, IL‐1 levels did not change significantly in comparison to baseline values, while LPS‐induced TNF production was apparently related to the degree of airway obstruction after CS treatment. In fact, TNF production by PBM from CS‐responsive subjects was significantly decreased in comparison to the levels determined before CS treatment, while PBM from CS‐resistant subjects produced the same cytokine levels regardless of CS treatment. The present study suggests that the determination of LPS‐induced TNF secretion by PBM could be used to confirm the effectiveness of CS treatment in asthma.
Effect of azelastine on the seasonal increase in non‐specific bronchial responsiveness to methacholine in pollen allergic patients. A randomized, double‐blind placebo‐controlled, crossover study
Authors: BALZANO G. , GALLO C. , MASI C. , COCCO G. , FERRANTI P. , MELILLO E. , SECCIA G.
Published: April 2006

Azelastine, a phthalazinone derivative, is a new potent, long acting, orally active anti‐allergic compound with particularly strong H1‐histamine receptor antagonistic effects which has been proven to possess in vitro and in vivo a number of anti‐inflammatory properties. The aim of the present study was to investigate whether azelastine would be able to prevent and/or reverse the seasonal increase in non‐specific bronchial responsiveness to methacholine in pollen allergic patients. Twelve atopic patients (5 males, mean age 31 years), skin positive exclusively to grass and/or Parietaria pollen extract, with rhinitis and mild asthma occurring in the spring for at least two years previously, were studied. After a 2 week run‐in period, oral azelastine, 4 mg twice daily, or placebo, was given for 2 weeks from the start of the pollen season, according to a randomized, double‐blind design. After 2 weeks, the treatments were crossed over. During both the run‐in and study periods, patients recorded rhinitis and asthma symptoms, additional antihistamine and bronchodilator drugs taken and peak expiratory flow measurements. A methacholine inhalation test was carried out on four occasions in each patient: before the run‐in period, before the start of the treatment, and at the end of the two 2 week treatment periods. Azelastine significantly reduced rhinitis symptoms and the need for antihistamine drugs, whereas asthmatic symptoms, use of bronchodilator drugs, peak flow recordings and bronchial responsiveness to methacholine were unaffected by the treatment. Compliance level and adverse side‐effects were not significantly different between active treatment and placebo. In the final subjective evaluation of the two treatments, eight out of 12 patients preferred azelastine. Thus, azelastine has been confirmed to be effective and safe in the treatment of seasonal allergic rhinitis. However, in our patients, we have not been able to demonstrate any anti‐asthmatic action of the drug.
Type I allergy induced by Limulus Amoebocyte Lysate (LAL)
Authors: EBNER C. , KRAFT D. , PRASCH F. , STEINER R. , EBNER H.
Published: April 2006

During the last few years Limulus Amoebocyte Lysate (LAL) has been extensively used to detect minimal amounts of endotoxins of Gram‐negative bacteria in products of the pharmaceutical industry, in food stuff, body fluids, house dust and room air. LAL is produced from cells of the haemolymph (amoebocytes) of the horseshoe crab (Limulus polyphemus), which respond with an extremely sensitive clotting system upon contact with endotoxins. In this study we demonstrate by typical case history, positive skin test and ELISA the occurrence of Type I allergy to LAL in a patient suffering from conjunctivitis and rhinitis at work.
Histamine content, synthesis and degradation in human nasal mucosa
Authors: ABE Y. , OGINO S. , IRIFUNE M. , IMAMURA I. , FUKUI H. , WADA H. , MATSUNAGA T.
Published: April 2006

Histamine content and enzyme activities of histamine metabolism, histidine decarboxy‐lase (HDC), histamine N‐methyltransferase (HMT) and histaminase (diamine oxidase, DAO) in human nasal mucosa were determined with a highly sensitive and specific fluorescent method which was combined with high performance liquid chromatography. Histamine content and HDC activity were determined in 10 specimens of nasal polyp, nine specimens of maxillary sinus and five specimens of inferior turbinate. HMT and histaminase activities were determined in 15 specimens of nasal polyp, nine specimens of maxillary sinus and five specimens of inferior turbinate obtained during surgical therapy. Histamine and activities of HDC, HMT and histaminase were detected in all specimens except the case of histaminase activity in one specimen of nasal polyp. The mean values of histamine content and activities of HDC, HMT and histaminase of human nasal mucosa were 137.3 nmol/g wet weight, 26.3 fmol/min/mg protein, 26.4 pmol/min/mg protein and 0.5 pmol/min/mg protein, respectively. Histamine content in the mucosal tissue of the maxillary sinuses was significantly higher than that of nasal polyps or inferior turbinates. There were no significant differences in HDC activities among three kinds of nasal mucosa. Activities of HMT and histaminase, including their kinetic constants (Km and Vmax values for histamine) indicated that HMT has a greater potential than histaminase for histamine degradation in the human nasal mucosa. The presence of these enzymes suggests that these activities constitute an important modulating factor in histamine mediated allergic and inflammatory reactions in human nasal mucosa.
In vivo allergenic activities of eleven purified members of a major allergen family from wheat and barley flour
Authors: ARMENTIA A. , SANCHEZ‐MONGE R. , GOMEZ L. , BARBER D. , SALCEDO G.
Published: April 2006

Eleven purified members of the α‐amylase/trypsin inhibitor family from wheat and barlcy that showed very different IgE‐binding capacities when previously assayed in vitro. were used in double blind in vivo diagnostic tests to further evaluate their allergenic activity. These tests were carried out in 31 patients who showed allergic scnsilization to wheat flour as veritied by skin test, RAST and challenge test. The three members of the protein family with highest igl; binding in vitro (the glycosylated subunits of tetrameric α‐amylase inhibitors CM16* from wheat and CMb* from barley, and the barley monomeric inhibitor BMAI‐1) were found to be the strongest allergens as indicated by‐skin sensitivity in prick tests.
Histamine‐releasing serum factors as a predictor of the outcome of insect sting reactions. Results from a multicentre study*
Authors: MOSBECH H. , SKOV P. STAHL , EBBESEN F. , EBBESEN K. , NORN S. , KRISTENSEN K. S.
Published: April 2006

Cord blood cells were incubated (passively sensitized) with sera from 27 patients with previous systemic reactions to insect stings. Histamine release (HR) from these cells was measured following exposure to venom extracts at increasing concentrations. The aim was to see whether this parameter could predict more efficiently than RAST and skin test the outcome of a subsequent re‐sting. Results showed that HR from passively sensitized cells tended to reflect skin sensitivity and specific IgE levels. If patients were not re‐stung during the follow‐up period, HR from the passively sensitized cells frequently decreased whereas an increase was seen (in 6/13) when using sera collected after re‐sting. In conclusion HR from passively sensitized cord blood cells could not satisfactorily predict re‐sting reactions in the serum donors.
Relation between bronchial responsiveness to methacholine and levels of IgE antibody against Dermatophagoides farinae and serum IgE in asthmatic children
Authors: TAKEDA K. , SHIBASAKI M. , TAKITA H.
Published: April 2006

The relation between non‐specific bronchial responsiveness and allergic sensitivity was evaluated in children with asthma. Bronchial responsiveness was determined by methacholine inhalation challenge test, and was expressed as a provocative concentration of methacholine causing a 20% fall in FEV1 (PC20). RAST titre for Dermatophagoides farinae and serum IgE level were evaluated as parameters of allergic sensitivity. When the PC20 vaiues, RAST titres for D. farinae. and serum IgE levels of 47 asthmatic children and 16 normal controls were compared. the asthmatic children had significantly lower PC20 values and higher D. farinae‐RAST titres and serum IgE levels than the normal controls. The correlation analyses in 47 asthmatic children have shown that there is no significant correlation between PC20 values and RAST titres for D. farinae (r= 0.04. P > 0.1) or between PC20 values and serum IgE levels (r= < 0.03 P > 0.1). These results suggest thaty although both bronchial hyperresponsiveness and allergic sensitizalion may be in some way related to one another in children with asthma, the magnitude of allergie sensilization does not influence the degree of the non‐specific bronchial responsiveness.
Rapid onset of action of levocabastine eye‐drops in histamine‐induced conjunctivitis
Authors: STOKES T. C. , FEINBERG G.
Published: April 2006

Levocabastine is a recently developed, potent H1‐receptor blocker intended for topical application. Seventeen healthy non‐atopic volunteers participated in this randomized, double‐blind, placebo‐controlled study undertaken to investigate the speed of onset of action, efficacy and tolerability of levocabastine eye‐drops after a histamine challenge. The degree of histamine‐induced ocular inflammation was found to be significantly less after administration of levocabastine compared with placebo. Furthermore, levocabastine was shown to be rapidly effective with an onset of action within 10 min. Levocabastine eye‐drops were well‐tolerated and no adverse reactions to the study drug were reported. Levocabastine eye‐drops appear to be a valuable therapeutic approach for the treatment of histamine‐mediated ocular allergies.
Absence of functional inhibition of cloned human β2‐adrenergic receptors by autoantibodies in asthmatic subjects
Authors: POTTER P. C. , VAN WYK L. , WHITE D. , DAKERS B. S. , CHUNG F. Z. , DOWDLE E. B.
Published: April 2006

Using guinea‐pig lung membranes and cloned human β2‐receptor adrenergic receptors the effects of whole serum, plasma, purified immunoglobulins and cellular activation products on β2‐adrenergic receptor ligand binding and function were investigated. Sera from 24 non‐asthmatic subjects and 115 asthmatics in different clinical categories were studied. There were no significant differences between antagonist ([125I] cyanopindolol) inhibition mediated by serum, plasma or by purified IgG when the asthmatics were compared with non‐asthmatics. There was also no inhibition of 10−6m isoproterenol stimulated cAMP release from L cells expressing human β2‐adrenergic receptors, by plasma, DEAE purified IgG fractions from asthmatics and non‐asthmatics, or by products of activated platelets or lymphocytes. Since we have no evidence that immunoglobulins from asthmatic subjects exert functional inhibition of human β2‐adrenergic receptors we conlcude that autoantibodies to the β2‐adrenergic receptors do not play an important functional role in the pathophysiology of asthma.
Suppressive effects of elimination diets on T cell responses to ovalbumin in hen's egg‐sensitive atopic dermatitis patients
Authors: SHINODA S. , KONDO N. , FUKUTOMI O. , AGATA H. , SUZUKI Y. , SHIMOZAWA N. , TOMATSU S. , YAMADA Y. , TAKEMURA M. , NOMA A. , ORII T.
Published: April 2006

We investigated the effect of elimination diets on T cell responses to ovalbumin (OA) in hen's egg‐sensitive atopic dermatitis (AD) patients. The proliferative responses of both peripheral blood mononuclear cells (PBMCs) and T cells with monocytes to OA decreased after elimination diets, but those to Candida albicans or phytohemagglutinin (PHA) did not decrease after elimination diets. The proliferative responses of CD4+ T cells with monocytes to OA decreased after elimination diets. In these patients, clinical symptoms of AD improved. These results indicate that T cells, especially CD4+ T cells, respond to food antigens in food‐specific lymphocyte responses, and that elimination diets may be able to initiate reduction of the responsiveness of food‐sensitive T cells, especially CD4+ T cells. Moreover, the surface marker phenotypes of the T cells responding to OA were analysed. Our results showed that CD4+CD45RA+ T cells tended to increase. The increase in circulating CD4+CD45RA+ T cells might function as systemic suppression against immune responses in the skin.
Influence of Kupffer cell phagocytosis blockade on the production of ovalbumin‐specific IgE and IgG1 antibodies in an experimental model
Authors: VIDAL C. , QUINTELA A. GONZÄLEZ , CUERVAS‐MONS V.
Published: April 2006

The aim of the present study was to evaluate the influence of Kupffer cell phagocytosis blockade (KCPB) on the production of reaginic (IgE) and non‐reaginic (IgGl) ovalbumin antibodies in an experimental animal model. Forty‐two female heterozygous Gunn rats were injected with two 100 μg doses of ovalbumin separated by 13 days, into jugular vein (Group I, n= 10), portal vein (Group II, n= 10), jugular vein with prior KCPB (Group III, n= 10) and portal vein with prior KCPB (Group IV, n= 12). KCPB was induced with gadolinium chloride (5 mg/kg body weight, 24 hr before each ovalbumin administration). Antibodies were determined by passive cutaneous anaphy‐laxis in sera obtained 12 days after the first dose of ovalbumin and 8 days after the second one. No antibodies were detected at any time in Group II. The maximum response was observed in Group IV in which, after two doses of ovalbumin, 100% of the animals presented IgGl and 58% presented detectable IgE antibodies. Differences between group IV and the other groups were statistically significant. This phenomenon does not seem to be due to a systemic effect of gadolinium chloride since humoral response was not increased in Group III with respect to Group I. It is concluded that the liver represents a barrier to IgE and IgG1 sensitization. KCPB not only eliminates this barrier, but also clearly increases antibody production to a protein antigen (ovalbumin) arriving at the liver through the portal vein.
Characterization of Aspergillus umbrosus carbohydrate antigens by biotinylated lectins and IgG response to mannan/mannoprotein antigens in patients with farmer's lung
Authors: KAUKONEN K. , SAVOLAINEN J. , VIANDER M. , TERHO E. O.
Published: April 2006

Twenly‐one different biotinylated lectins were used to recognize the carbohydrates of Aspergillus umbrosus, one of the most common microbes that patients with farmer's lung in Finland are exposed to. The glycoprotein fraction of A. umbrosus was bound especially well by Concanavalin A and consisted mainly of carbohydrates mannose and glucose. The carbohydrate fraction of A. umbrous antigens were isolated from the crude extract of A. umbrosus with Con A‐Sepharose affinity chromatography. Serum IgG antibodies to A. umhrosus mannoprotein fraction were determined in 57 patients with farmer's lung, 10 asymptomatic exposed farmers and 10 healthy controls by enzyme linked immunosorbent assay. Candida albicans and Saccharomyces cerevisiae mannan were used as controls. Patients had high levels, whereas asymptomatic exposed farmers showed moderate, and healthy controls low levels, of IgG antibodies to the A. umbrosus mannan/mannoprotein fraction. There were no significant differences in the mean antibody levels between the patients and controls against C. albicans or S. cerevisiae mannose fraction although in all groups more antibodies were detected against C. albicans than S. cerevisiae.