bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunological study of Melkersson‐Rosenthal syndrome. Lack of response to food additive challenge
Authors: MORALES C. , PEÑARROCHA M. , BAGÁN J. V. , BURCHÉS E. , PELAEZ A.
Published: April 2006

A study was made of six patients with Melkersson‐Rosenthal syndrome (MRS) to establish the aetiological role of foodstuffs and/or additives and the possible associated immunological alterations. In all cases Melkersson‐Rosenthal syndrome (MRS) was diagnosed both clinically and histologically, excluding other causes of orofacial granulomatosis (OFG). A detailed study of possible triggering factors was performed in all patients. Blood analysis, x‐rays and cultures, were always within normal limits, with the exception of the finding of circulating immune complexes (CICs) in three patients with facial palsy associated, and C‐reactive protein positivity in two patients who presented persistent labial oedema. All patients were subjected to skin‐prick tests with common inhalant allergens and with foods when sensitization to foods were suspected, and patch tests with European Standard Series and pastry components, organic dyes, perfumes and fragrances series. The results were negative in all cases. When asymptomatic, the patients were subjected to a double‐blind oral challenge, under placebo control, with additives (monosodium glutamate, tartrazine, sulfites, erythrosine, paraoxybenzoate, sodium benzoate, lactose, aspirin, and annate), which was again negative. In no case did the patients refer the appearance of outbreaks with exposure to foods or contactants, and the course of the disease was unaffected by exclusion diets and the elimination of contactants. To conclude, we observed no sensitization to foods, additives or contactants in our patients. Likewise, there were no antecedents of atopy or hereditary predisposition related to the aetiopathogeny of MRS. The significance of the CIC encountered only in patients with facial paralysis remains to be established, due to the limited number of patients studied.
Horse gammaglobulin‐induced thrombocytopenia in anaphylaxis involving sequestration and activation of platelets
Authors: LEIR S.‐H. , CHEN S.‐H. , LEI H.‐Y.
Published: April 2006

Thrombocytopenia as well as hemoconcentration and leukopenia followed by leuko‐cytosis were induced after HoGG challenge on HoGG‐sensitized mice. Thrombocytopenia was induced within 2min and sustained for 1 day. HoGG‐induced thrombocytopenia was not observed until day 10 after sensitization; mice challenged with HoGG dose ≥10μg developed thrombocytopenia. Two types of thrombocytopenia were observed in appropriately sensitized mice. HoGG induced thrombocytopenia at 2min and 60 min, whereas, α‐macroglobulin induced thrombocytopenia at 2min, the platelet count of which returned to normal within 60min. Poly (Glu60Ala30 Tyr10) did not induce thrombocytopenia at 2min or 60min. The tracing study by 3H‐serotonin labelled platelets demonstrated the 2min‐sequestration of platelets in lungs or livers. The HoGG‐induced sequestration of platelets at 2min was blocked by high dose heparin or Cobra Venom factor. Platelet activation at 60min was partially inhibited by dexamethasone, rhodostomin synthetic peptide 45–59, or platelet activation factor antagonist (WEB 2086). Furthermore, the thrombocytopenia could be transfered by heat (56°C, 4h)‐treated immune sera. This suggests that HoGG‐induced, non‐IgE‐mediated thrombocytopenia in anaphylaxis involves sequestration and activation of platelets. The sequestion in lungs occurs within 2min and can be inhibited by high dose heparin or Cobra Venom factor. The activation of platelets involves platelet activation factor, and fibrinogen receptor.
Experimental intestinal hyper sensitivity. Effect of four anti‐allergic drugs on protein uptake, permeability to sugars and mucosal mast‐cell activation
Authors: TURNER M. W. , BOULTON P. , STROBEL S.
Published: April 2006

The ability of four drugs with anti‐allergic action to modulate the uptake of bystander protein, lactulose/rhamnose permeability ratios and mast cell activation was studied in rats presensitized with egg albumin in alum and challenged intraduodenally with the same antigen. Beclomethasone dipropionate (BDP) and nedocromil both significantly reduced the uptake of the bystander protein, bovine serum albumin (P < 0.002 and P>0.02 respectively). BDP also significantly reduced sugar permeability (P<0.1). In animals with elevated lactulose/rhamnose permeability ratios we confirmed our earlier observation of a significant correlation between levels of the specific mucosal mast cell protease Rat Chymase II (RChyII‐previously known as RMCPII) and the sugar ratios. None of the drugs had any influence on the levels of mast cell protease II released following challenge and there was no correlation between the histological light microscopic appearance of the mast cells and the experimental treatment administered. Our results suggest that in the gut the pharmacological effect of anti‐allergic drugs may be complex. Some, such as nedocromil, appear to act only on the mechanisms underlying increased protein uptake whereas others, such as BDP, appear to abrogate both increased protein uptake and increased sugar permeability.
Localization of major allergens in the dust mite Lepidoglyphus destructor with confocal laser scanning microscopy
Authors: VAN HAGE‐HAMSTEN M. , OLSSON S. , EMILSON A. , HÄRFAST B. , SVENSSON A. , SCHEYNIUS A.
Published: April 2006

The dust mite Lepidoglyphus destructor is the dominating source of allergens giving rise to asthma and rhinitis among farmers. In a previous study of the localization of allergens in L. destructor we demonstrated that the 39 kDa allergen is associated with digestion. Here we describe the localization of the principal 15 kDa allergen and the high molecular weight allergen complex (79 and 93 kDa) in L. destructor with confocal laser scanning microscopy (CLSM). Cryostat‐cut sections of mite bodies and faecal pellets were probed with mouse monoclonal antibodies (MoAbs) raised against the allergens. The 15 kDa allergen disclosed labelling of the mite body and most of the faecal pellets but left the exoskeleton unlabelled. The binding was widespread, and most intense in the mouth region. However, some staining was also observed around the gastrointestinal tract. In contrast, the 79 and 93 kDa allergen complex stained the exoskeleton and the front part of the mile. Interestingly, we detected no labelling of the faecal pellets with the MOAb against the 79/93 kDa allergen. The study indicates that the 15 kDa allergen is associated with the digestive tract whereas the function of the 79 and 93 kDa allergen complex remains to be elucidated.
Inhalant allergen‐specific T‐cell reactivity is detectable in close to 100% of atopic and normal individuals: covert responses are unmasked by serum‐free medium
Authors: UPHAM J. W. , HOLT B. J. , BARON‐HAY M. J. , YABUHARA A. , HALES B. J. , THOMAS W. R. , LOH R. K. S. , O'KEEFFE P. T. , PALMER L. , LE SOUEF P. N. , SLY P. D. , BURTON P. R. , ROBINSON B. W. S. , HOLT P. G.
Published: April 2006

Background
It is widely held that in vitro T cell responses to allergens are more prominent in atopic than in normal individuals, though this conclusion is based upon culture techniques which fail to detect proliferative responses in a significant minority of atopies and many normals.

Objectives: Study allergen‐specific proliferative responses of T cells cultured in serum‐free medium (SFM). Examine associations between atopic status, age and T cell reactivity.

Methods
Initially, peripheral blood mononuclear cells were stimulated with allergens or antigens in SFM, and compared with cells cultured in RPMI + 10% fetal calf serum or human AB serum. Subsequently, T cell reactivity was studied in 34 adults (20–49 years), 27 children (2–13 years), and 19 infants (≤ 10 weeks) using SFM alone. Results Compared with serum‐supplemented medium, SFM enhanced net T cell proliferation, both in bulk culture and when cloning at limiting dilution. In many subjects, SFM unmasked T cell reactivity to allergens which was not otherwise evident, and lowered the threshold allergen levels required for in vitro T cell triggering. For most allergens, T cell proliferative responses did not differ between adults who had specific IgE, and those who did not. The most vigorous responses observed were to ubiquitous inhalant allergens, which stimulated T cells from close to 100% of adults and children, and over 60% of infants. In contrast, responses to the ‘vaccine’ antigen tetanus toxoid were completely absent in the latter age group, but present in the majority of adults and children.

Conclusions
These findings suggest that the extent of active T cell recognition of environmental allergens has been hitherto underestimated, and further that these responses may frequently be initiated in very early life. Additionally, these findings reinforce the notion that qualitative (as opposed to quantitative) variations in specific T cell reactivity ultimately determine allergen responder phenotype.
Density of eosinophils reflects activity of disease in allergic asthmatic children
Authors: KROUWELS F. H. , KERSTENS L. C. M. , VAN DER MAAREL H. W. M. , DEGENHART H. J. , NEIJENS H. J.
Published: April 2006

Background Low density eosinophils are more prominent in asthmatic patients compared with healthy subjects, LDH are metabolically more active and produce more tissue‐injuring and spasmogenic proteins than normal cosinophils.

Objective and methid With a method providing information about eosinophils of 12 different densities we were able to study eosinophil density characierislics in 24 young patients in detail with allergic asthma in a stable phase, and in 21 patients after a bronchial allergen challenge.

Results Study of the eosinophil density profile of patients and healthy controls revealed two density populations. Patients had more low density eosinophils than controls. In the patients eosinophil density characteristics and in particular the number of low density eosinophils correlated strongly with both FEV1% predicted (ρ=−0.66, P < 0.001) and REV1/FVC (ρ=−0.47, P < 0.01) as well as with bronchial responsiveness to histamine (ρ=−0.68, P < 0.001) and house dust mite (ρ=−0.37, P < 0.05). Allergen induced bronchial reactions were associated with an increase in the number (P < 0.001) and percentage (P < 0.05) of low density eosinophils. A selective rise in the number of eosinophils collected from fractions with a low density accounted for the observed rise in the total number of eosinophils, Density changes did not differ between patients with an isolated early reaction and patients with both an early and a late reaction, nor was there a relation between the severity of the late reaction and the shift in eosinophil density.

Conclusion In conclusion, peripheral blood eosinophil density characteristics and in particular numbers of low density eosinophils are closely related with indicators of the asthma severity under stable conditions. Allergen inhalation induces a further shift towards lower density suggesting additional activation of the eosinophils.
Influence of culture period on the allergenic composition of Pityrosporum orbiculare extracts
Authors: ZARGARI A. , DOEKES G. , DIJK A. G. VAN IEPEREN‐VAN , LANDBERG E. , HÄRFAST B. , SCHEYNIUS A.
Published: April 2006

Background Previous characterization studies of Pityrosporum orbiculare allergens have led to contradictory results. In immunoblotting studies a range of IgE‐bitiding proteins of 10–100kDa have been identified. In another study, however, the IgE‐binding structures were claimed to be associated with high‐molecular‐weight polysaccharides or glycoproteins, presumably mannans or mannoproleins. Objective In the present study the reasons for these discrepancies were investigated.

Methods P. orbiculare preparations were compared in IgE ELISA and IgE‐inhibilion ELISA, as well as in immunoblotting with sera from atopic dermatitis patients.

Results It was inferred that variations in the period of in vitro culture of P. orbiculare constituted the most important factor determining the different compositions of the resulting yeast cell extracts. After 2 days of culture a wide range of allergen if proteins was present but upon more prolonged culture (>4 days) most proteins of 10‐ 100kDa were lost. Accordingly, the protein concentration of the extracts gradually declined from 40% to 25% between days 4 and 15 of culture. On the other hand, the carbohydrate content remained fairly constant (approximately 30%). Using inhibition ELISA it was demonstrated that the high‐molecular‐weight glycoproleins or poly‐saccharides presumably involved in most of the IgE‐binding capacity in extracts from old cultures, were also present in comparable concentrations in all extracts tested, even after culture for only 2 or 4 days.

Conclusion Preparations obtained from the exponential phase of yeast cultures (2–4 days old), should preferably be used in studies of the IgE response to P. orbicularc.
Differences in titres of IgE, IgG4 and other IgG subclass anti‐ Der p 2 antibodies in allergic and non‐allergic patients measured with recombinant allergen
Authors: TAME A. , SAKIYAMA Y. , KOBAYASHI I. , TERAI I. , KOBAYASHI K.
Published: April 2006

Background
The mite allergens are recognized as major causes of allergic disease such as bronchial asthma, allergic rhinitis and atopic dermatitis. The functions of allergen‐specific IgG subclass antibodies are not defined.

Objective
In order to clarify the relationship between IgE and IgG subclasses, we examined scrum levels of the Dermatophagoides pteronyssisus group 2 (Der p 2)‐specific antibodies of IgH. IgG total and IgG subclasses in children with mite allergy.

Methods
We prepared a recombinant Der p 2 fusion protein and examined serum levels of Der p 2 antigen‐specific antibodies by enzyme‐linked immunosorbent assay (FLISA) systems developed in our laboratory using a recombinant Der p 2 as target antigen. Sera from 240 children with mite allergy and 25 controls were measured.

Results
The serum levels of specific IgE and, to lesser degree, lgG4 were higher in allergic children than non‐allergic controls, while in the levels of the other IgG subclasses there was no difference between the two groups. There was no correlation between levels of specific IgF and IgG4 or in those between specific IgG4 and other IgG subclasses.

Conclusion
Results indicate that the induction of Der p 2‐specific lgG4 in allergic diseases is independent to IgE as well as other IgG subclasses.
Effect of cetirizine and prednisolone on cellular infiltration and cytokine mRNA expression during allergen‐induced late cutaneous responses
Authors: TABORDA‐BARATA L. , JACOBSON M. , WALKER S. , NJUKI F. , YING S. , RANDEV P. , DURHAM S. R. , ICAY A. B.
Published: April 2006

Background
The ability of cetirizine to inhibit eosinophil infiltration into the sites of allergen‐induced cutaneous late‐phase reactions is controversial. A previous skin biopsy study gave negative results with 15 mg of cetirizine as a single dose.

Objective
To confirm these findings we have used cetirizine (30 mg daily) for 5 days and compared the results with prednisolone (20 mg daily for 5 days) as a positive control. The efTect of these agents on mRNA positive cells for inlerleukin‐3 (IL‐3). interleukin‐4, interleukin‐5 and granulocytc/macrophage‐colony stimulating factor (GM‐CSF) was also evaluated.

Methods
A double‐blind, placebo‐controlled cross‐over study (n= 12) was followed. After each treatment 30 biological units (BUs) of Dermatophagoides pteronyssinus or Phleum pratense were injected inlradermally and the early (15min) and late‐phase response sizes (6 and 24h) were measured. Skin biopsies were taken at 24h for immunocytochemistry and in situ hybridization.

Results
Cetirizine but not prednisolone inhibited the early‐phase response (37%, p = 0.004). In contrast prednisolone. but not cetirizine, significantly inhibited the size of the late‐phase reaction at 24 h (70%. P = 0.021). This was associated with significant decreases in tolal (MBP+) and activated (EG2+) eosinophils (P= 0.019 and 0.014, respectively), as compared with placebo. There were also clear but non‐significant reductions in interleukin‐3, interleukin‐4, interleukin‐5 and granulocyte/macrophagc‐colony stimulating factor mRNA+ cells.

Conclusion
Prednisoione, but not cetirizine. inhibited both the magnitude of the allergen‐induced late‐phase response and the accompanying local eosinophil infiltration. These corticosleroid effects were associated with a reduction in cells expressing mRNA for ‘TH2‐type’ cytokines.
An allergenic polypeptide representing a variable region of hsp 70 cloned from a cDNA library of Cladosporium herbarum
Authors: ZHANG L. , MURADIA G. , DE VOUGE M. W. , RODE H. , VIJAY H. M.
Published: April 2006

Background
Extracts of Clado.sporium herharum, a major source of fungal aeroaller‐gens. exhibit a complex profile of IgE‐binding proteins. Yields of conventionally purified allergens from this mold have been insufficient to permit further molecular analyses.

Objective
To enhance and simplify the purification of allergens from C. herbarum, we have sought to use recombinant DNA techniques to clone, identify and bacterialiy express immunoselected C. herbarum allergens.

Methods
We constructed a cDNA library in AZAP II using mRNA isolated from C. herbarum. From this library, phage clones encoding a new allergen were immunoselected using pooled human atopic IgE. The cloned cDNA was excised from the phage vector asa recombinant pBluescript II SK‐phagemid und sequenced. Expression of the recombinant allergen was carried out in E. coli XLl‐blue transformants of the phagemid. Bacterial lysates from cells induced to express the cloned allergen were immunoblotted and probed with individual human atopic IgEs.

Results
The cDNA clone encodes a 278 amino acid polypeptide homologous to the C‐terminal portion of 70 kDa heat shock protein (hsp 70). The polypeptide possesses features common to other hsps 70. i.e. a similar hydropathic profile and a variable C‐terminal region with conserved sequence at the very C‐terminus. Binding of the recombinant peptide to IgE from 38% of atopic sera or plasma from individuals allergic to C. herbarum was demonstrated.

Conclusion
These results indicate that amino acid substitutions are relatively conserved even in the variable C‐tcrminal regions ofhsp 70 species. Thus, this study should draw attention to the possibility of induction of anaphylactic responses in a sensitized individual when hsp 70 from any pathogenic species is administered for vaccination.
Diversity of allergens causing occupational asthma among cereal workers as demonstrated by exposure procedures
Authors: ALVAREZ M. J. , TABAR A. I. , QUIRCE S. , OLAGUíBEL J. M. , LIZASO M. T. , ECHECHIPíA S. , RODRíGUEZ A. , GARCíA B. E.
Published: April 2006

Background
Occupatiotial asthma among cereal workers is frequently due to cereals but other allergens can also be responsible.

Objective
We evaluated the allergens causing occupational asthma in the patients who had been diagnosed in our Department during the last 5 years. Specific bronchial reactivity to the implicated allergens was assessed and compared by standardized bronchial challenge.

Methods
Twenty‐one patients (12 bakers, three millers and six farmers) were studied. We carried out in vivo tests (skin and challenge tests) and in vitro tests (specific IgE measurement) with cereals, enzymes, soyabean, storage mites and egg. A definitive diagnosis was established by means of specific bronchial provocation tests (BPT), except in three patients in whom it could not be carried out due to the severity of their asthma. In these cases the causative agent had to be determined by means of conjunctival challenge.

Results
Cereals were the main sensitizers among bakers (75%) and farmers (66%). Bakers were also sensitive to a‐amylase (41%) and soyabean (25%), and farmers, to soyabean (33%) and storage mites (33%). Occupational asthma was due to cereals, soyabean and storage mites among millers.

Conclusions
Besides cereals, other allergens such as enzymes, leguminous, egg and storage mites can be the causative agents of occupational asthma among cereal workers. Sensitization to different allergens in the different jobs is very likely due to differences in exposure.
Wheat antigen exposure assessment for epidemiological studies in bakeries using personal dust sampling and inhibition ELISA
Authors: HOUBA R. , VANRUN P. , HEEDERIK D. , DOEKES G.
Published: April 2006

Background
Asthma in bakery workers caused by exposure to wheat flour proteins is an important occupational health problem. Until recently, gravimetric dust measurements were the only available technique for quantitative exposure assessment in bakeries. However, it is questionable whether dust levels are a good exposure parameter or only give a crude approximation of the actual flour allergen concentration.

Objective
In the present study we have investigated a method to measure wheat flour antigens with immunochemical methods.

Methods
Wheat flour antigens were measured in 449 personal dust samples taken in bakeries, using enzyme‐linked immunosorbent assay (ELISA) inhibition and an anti‐wheat lgG4 serum pool. Western‐blotting was performed to compare the wheat flour proteins detected by IgE and IgG4.

Results
Electrophoresis and immunoblotting showed that many wheat flour proteins can bind lgG4 and IgE, but also a reasonable similarity in major allergens detected by our lgG4‐serum pool and IgE‐positive sera. Inhibition tests showed some cross‐reactivity with some cereal species, but not with other ingredients used in bakeries. In bakeries, large differences in personal airborne flour levels were found between occupational titles. Eor several groups clear differences in wheat antigen exposure levels existed, where no difierences in dust exposure levels could be found. The relationship between dust and wheat antigen exposure varied considerably, depending on the specific bakery occupation, the size of the bakery, and the type of product produced by the bakery. This study also shows that personal sampling of wheat antigens is possible on a large scale and can be used for epidemiological field studies.

Conclusion
Measurement of airborne wheat antigens in bakeries is a more specific and sensitive measurement tool than measuring dust samples, and will probably be essential for epidemiologic field studies focusing on exposure‐response relationships.
Diminution of T cells with γδ receptor in the peripheral blood of allergic asthmatic individuals
Authors: CHEN K.S. , MILLER K. H. , HENGEHOLD D.
Published: April 2006

Background
T cells are shown to regulate allergy and asthma. They are heterogeneous by virtue of surface receptor subtypes (αβ and γδ receptors), however, their pathophysiological roles in asthma are unclear.

Objective and methods
The present study employed flow cytometric analysis to compare the size of T‐cell subsets and eosinophils present in the peripheral blood of healthy, allergic and allergic asthmatic individuals.

Results
Current results demonstrated that the percentages of γδ T cells declined from 4.1% in healthy to 3.2% in the allergic subjects and to a significantly lower (P= 0.01) 2.4% in allergic asthmatics. The absolute numbers of circulating γδ T cells also were diminished in a similar fashion such that healthy individuals had a significantly higher mean cell count (91.8 ± 103/mL) than did allergic asthmatics (47.8 ± 103/mL) (P= 0.0266). In contrast, αβ T cells were comparable in the healthy, allergic, and allergic asthmatic populations (65.3%, 65.8% and 69.4%, respectively); the differences were not statistically significant. On a populational basis, the proportion of individuals having both γδ T cells y± 4.1% and eosinophils ≥2.1% was the lowest in the healthy population (30.8%), but was elevated in the allergic group (85.7%) as well as in the allergic asthmatic group (86.6%). However, on an individual basis, those who had reduced γδ T cells did not have consistently higher eosinophil counts or IgE level. Conclusion It was concluded that no significant correlation existed between the levels between γδ T cells and eosinophils or between γδ T cells and IgE present in the peripheral blood. This report, for the first time, documents that allergic asthmatics have reduced γδ T cells with reciprocally elevated eosinophil numbers in their peripheral circulation. However, it does not indicate that the reduction of γδ T cells directly correlates with the predominance of eosinophils of IgE levels in the diseased populations. The pathophysiological role of γδ T cells in allergic diseases awaits further exploration.
Immunoglobulin E in acute phase response to surgical stress
Authors: SZCZEKLIK A. , JAWIEN J.
Published: April 2006

Background
A few years ago we reported that acute myocardial infarction is associated with a distinct, transient rise in serum imtnunoglobulin (Ig)E.

Objective
We wondered whether this response is specific only for myocardial infarction or whether it reflects a more generalized phenomenon, precipitated by tissue injury.

Methods
We carried out a large prospective study on 149 patients undergoing various surgical procedures. These were the patients undergoing coronary artery bypass surgery, who did (n=39) or did not (n=42) develop perioperative myocardial infarction, patients subjected to various thoracic operations (n= 33), eholecystectotny (n = 17) or repair of the inguitial hernia (n=18). Thirty healthy volunteers fortned the control group.

Results
In all groups studied, except the control group, serum IgE levels began to rise shortly after the operation, reached a peak by the fifth postoperative day. and then gradually declined. This was in striking contrast to serurn imnunoglobulin G which soon alter the operation became markedly) depressed, and took more than a week to return to preoperative values. Specific IgE. against common aeroallergens, measured in a subgroup of patients with initially high total serum IgE, showed no uniform pattern of change.

Conclusion
Surgical operations are accompanied by a characteristic, transient rise in serum IgE concentration. Immunoglobulin E might be involved in acute phase response to tissue injury.
Latex allergy: allergen identification in Hevea braziliensis fractions by immunoblotting
Authors: MUGUERZA J. , CAPO C. , PORRI F. , JACOB J.‐L. , MEGE J.‐L. , VERVLOET D.
Published: April 2006

Background
Latex is the cause of several clinical symptoms of allergy, but the identification of allergens is not completely known.

Objective
The aim of this report was to study the immunoreactivity of puritied stable latex fractions from Hevea braziliensis.

Methods
We purified the cytoplasm of Hevea braziliensis and obtained three fractions: latex particles (LP), lutoids (L) and cytosolic serum (CS). Using Western blot, specific IgE directed to latex allergens was found in 80 patients with latex allergy.

Results
Five major groups of allergens migrating as 14, 25, 29, 37–45 and 50kDa were recognized. They were unequally distributed within the latex fractions: 37–45 kDa proteins were essentially recognized in CS and LP, whereas 14 and 29 kDa proteins were mainly labelled in the L fraction. As a control, aqueous glove extracts exhibited a more restricted pattern of reactivity, because only 14 and 29 kDa proteins were recognized by patient sera. The pattern of reactivity was not correlated with specific IgE levels, but sera from patients suffering from spina bifida reacted specifically with the minor protein of 25 kDa located in LP.

Conclusions
The present results show that latex allergic patients recognize several allergens which are differently distributed in subcellular fractions extracted from H. braziliensis and aqueous GE. The L fraction and GE were enriched in low molecular weight proteins and apparently contained the same allergens.
Effects of prednisolone on bone turnover in patients with corticosteroid resistant asthma
Authors: LANE S. J. , VAJA S. , SWAMINATHAN R. , LEE T. H.
Published: April 2006

Background
Although corticosteroid resistant (CR) bronchial asthma is associated with impaired in vitro and in vivo glucocorticoid (GC) responsiveness in mononuclear cell function, it is not known whether this is a cell specific phenomenon or whether these patients are yt similar risk of glucocorticoid side‐effects as corticosteroid‐sensitive (CS) subjects. In order to address this question we have examined the effects of GCs on biochemical indices of bone turnover in vivo.

Methods
Six CS and six CR subjects received prednisolone 40 mg orally at 09.00 daily for 5 days. At 08.30 on day 1 and day 6 a fasting blood was taken for estimation of serum osteocalcin, tartrate resistant acid phosphatase (TRAP) and alkaline phosphatase (ALP: total and bone isoenzyme), and a urine sample taken for estimation of free deoxy‐pyridinoline crosslinks. TRAP and ALP were measured by a colorimetric method and osteocalcin and deoxy‐pyridinoline crosslinks by ELISA.

Results
Serum osteocalcin (nmol/L) decreased from 1.39 ± 0.O9 (mean ± SEM) to 1.14 ± 0.07 (P= 0.023) and from 1.19 ± 0.05 to 0.96 ± 0.1 (P= 0.037) in the CS and CR groups, respectively. There was no difference between baseline and post‐treatment levels in either group. Serum total ALP (units/L) decreased from 178 ± 7 to 168 ± 4 (P= 0.02) and from 195 ± 22 to 175 ± 16 (p=0.04) in ihe CS and CR groups, respectively. There was no difference between baseline and post‐treatment levels in either group. There was no significant elevation of deoxy‐pyridinoline crosslinks or TRAP and no suppression of bone ALP in either group.

Conclusion
We suggest that CR asthmatics may be equally at risk from the metabolic side‐effects of glucoeortieoids.
Domestic allergens in public places II: dog (Can f 1) and cockroach (Bla g 2) allergens in dust and mite, cat, dog and cockroach allergens in the air in public buildings
Authors: CUSTOVIC A. , GREEN R. , TAGGART S. C. O. , SMITH A. , PICKERING C. A. C. , CHAPMAN M. D. , WOODCOCK A.
Published: April 2006

Background
Sensitization and exposure to indoor allergens are the major risk factors for asthma. It is possible that significant exposure to domestic allergens occurs outside the home.

Objectives
To investigate the levels of Can f 1 and Bla g 2 in the dust from carpeted floors and upholstered seats in public buildings and public transport and the airborne concentrations of Der p 1, Fel d 1, Can f 1 and Bla g 2 in schools and offices.

Methods
Can f 1 and Bla g 2 were measured in the dust collected by vacuuming a I m2 area of carpet, as well as upholstered seats in five schools, six hotels, four cinemas, six pubs, three buses and two trains. Dust was also collected from the bedroom carpet, living room carpet, mattress and sofa in 20 homes with and 20 homes without a dog in the same area. Personal airborne sampling (2 L/min) was conducted for 8 h in offices (n= 16) and classrooms (n= 9). In addition, airborne samples in schools were collected using a high volume pump (60 L/min) for 1 h in three classrooms immediately after the children vacated the school. Can f 1, Bla g 2, Der p 1 and Fel d 1 were assayed using a two–site monoclonal antibody–based ELISA.

Results
Can f 1 was detected in all dust samples from public places, ranging from 0.2 to 52.5 μg/g, Significantly higher levels were found in upholstered scats (geometric mean – GM 9.4 μg/g) than in carpets (GM 1.5 μg/g; P < 0.001), and levels of Can f 1 > 10 μg/g were found in 40% of upholstered seats in public places. Can f 1 was significantly higher in upholstered seats in public places than in sofas in homes without a dog (GM 1.8 μg/g; P < 0.001). Detectable levels of Bla g 2 were found in all of the schools (GM 2.4 U/g, range 0.8–4.4 U/g). Bla g 2 concentration greater than 2U/g (provisional threshold level representing risk of sensitization) was measured in 65% of the classrooms sampled. Der p 1 and Bla g 2 were below the detection limit in all airborne samples. However, airborne Fel d 1 and Can f 1 were detected in schools and offices, albeit in low concentrations.

Conclusions
Upholstered seats from public places constitute a reservoir for the accumulation of dog allergen, and a source of exposure to Can f 1 inside public buildings or on public transport. Exposure to cockroach allergens in schools may be important for cockroach sensitized asthmatic children.
Sublingual versus injective immunotherapy in grass pollen allergic patients: a double blind (double dummy) study
Authors: QUIRINO T. , IEMOLI E. , SICILIANI E. , PARMIANI S. , MILAZZO F.
Published: April 2006

Background
Injective immunotherapy is a well‐known and recognized treatment for allergic diseases, but its safety has been questioned during recent years. Alternative administration routes have been proposed and there is a growing interest and experience in sublingual therapy. The safety of alternative routes is nonetheless a real advantage, so long as it is not counterbalanced by a loss of clinical benefit.

Objective
We have compared the efficacy of the same biologically standardized grass pollen extract administered through the injective or the sublingual route, in a group of 20 patients followed for two pollen seasons.

Methods
Both therapies were administered for 12 months according to a double‐blind (double‐dummy) plan; at the end of the trial the cumulative dosage of the sublingual therapy was 2.4 times higher than that of the injective therapy. Data about skin reactivity, symptoms and drugs scores during the pollen season, as well as total specific IgG and specific IgG4, during and after the trial, were obtained.

Results
Our data show that sublingual and injective therapy are equally effective according to subjective clinical parameters, with a statistically highly significant reduction of symptoms and drugs (P= 0.002 for symptoms and drugs in SLIT‐treated patients; P= 0.002 for symptoms and P= 0.0039 for drugs in patients given injections). On the other hand, objective parameters (total specific IgG, specific IgG4, skin reactivity) changed only in patients treated with active injective therapy, with P < 0.001, P < 0.001 and P= 0.021, respectively.

Conclusions
The discrepancies observed could be interpreted as a consequence of different mechanisms of action of the two therapies or to the lack of close relationships between the clinical and the objective parameters which were considered here.
Occupational allergy due to spider mites: Tetranychus urticae (Koch) and Panonychus citri (Koch)
Authors: BURCHES E. , PELAEZ A. , MORALES C. , BRASO J.V. , ROCHINA A. , LOPEZ S. , BENITO M.
Published: April 2006

Background
Allergy to both house dust and storage mites is well established, but information about other species of mites is seant.

Objective
One hundred and fifty patients directly exposed to an occupational environment were studied to assess whether spider mites (Tetranychidue) caused their allergic symptoms. We also studied a group of 50 patients from an urban environment, who were not occupationally exposed to spider mites, with a strong sensitization to Dermatophagoides pteronyssinus (RAST class 4).

Methods
Case history (including questions about work‐related symptoms), skin tests, RAST and conjunctival provocation tests were performed in both groups using Tetranychus urticae and Panonychus citri extracts as allergens. Cross‐reactivity between spider mites and D. Pteronyssinus was determined by RAST inhibition.

Results
Fifty‐four of 150 rural workers were positive to Tetranychidae and in all cases there was an associated sensitization to D. pteronyssinus. All individuals belonging to the urban group were positive to spider mites. RAST inhibition demonstrated a significant cross‐reactivity between Tetranychidae and D. pteronyssinus. Five of fifty‐four rural workers sensitized to spider mites developed symptoms only when they handled plants or fruits infested with spider mites and they became asymptomatic when exposure ceased.

Conclusion
In the rural population studied. 36% of workers were found to be sensitized to spider mites and 10% had symptoms associated with occupational exposure. Since specific IgE antibodies to spider mites could not be detected in the absence of the specific IgE antibodies to D. pteronyssinus, and as all the affected workers were RAST positive to D. pteronyssinus, prior sensitization to house dust mites may be a risk factor for occupational allergy to spider miles.
Cytokine production in children outgrowing hen egg allergy
Authors: NOMA T. , YOSHIZAWA IZUMI , AOKI K. , YAMAGUCHI K. , BABA M.
Published: April 2006

Background
Approximately 40 to 80% of egg‐allergic children outgrow egg allergy after 2 to 5 years.

Objective and methods
To detail the immunologic mechanisms involved in the development of tolerance to egg proteins, the bahince between interleukin 4 (IL4) and interferon‐γ (IFN‐γ) synthesis in patients with active atopic dermatitis allergic to hen egg and in those outgrowing hen egg allergy was evaluated.

Results
A marked increase in IL4 and a decrease in IFN‐γ synthesis by peripheral blood lymphocytes following ovalbumin (OVA) specific in vitro stimulation was observed in active atopic dermatitis. In contrast, OVA‐induced IL4 synthesis in patients in remission was comparable to that in normal individuals. An intriguing finding was higher production of IFN‐γ by lymphocytes from ovalbumin‐inscnsitive patients in remission as compared to normal individuals following antigen stimulation. although cell proliferation in OVA‐stimulated lymphocytes was reduced in patients during remission.

Conclusion
OVA antigen may be capable of inducing a population of Thl‐type cells to produce cytokines such as IFN‐γ, resulting in suppression of Th2‐type responses, i.e. IL4 secretion. We speculate that the changes in the balance of relevant antigen‐induced cytokine synthesis seen in such patients may be causally associated with the improvement in their clinical status.
Isotypic analysis of grass pollen‐specific antibodies in human plasma. 4. Biological activity of allergen‐specific and autoanti‐IgE antibody fractions on basophil histamine release
Authors: BATARD T. , WEYER A. , LAROZE A. , ICKOVIC M.‐R. , DAVID B. , PELTRE G. , BASUYAUX B.
Published: April 2006

Background
Blocking antibodies are defined as antibodies that compete with IgE for binding to allergens due to their specificity for those allergens. Thus, they may inhibit allergen‐induced basophil and mast cell IgE‐dependent mediator release both in vivo and in vitro.

Objective
The present study was designed to evaluate the ability of antibodies isolated from human plasma samples on a Dactylis glomerata (Cocksfoot) pollen affinity‐column to inhibit the Dactylis pollen‐induced histamine release from human basophils (BHR) in vitro.

Methods
Antibodies from Ig pools containing either high or low IgG4 anti‐Dactylis pollen were purified on a Dactylis pollen affinity‐column and then separated on an anti‐human Igl column. Obtained Ig fractions were incubated for 30 min with Dactylis pollen allergens prior to incubation with basophils from Dactylis pollen‐allergic donors. Cell supernatants were assessed for histamine content and the inhibition of BHR was calculated.

Results
Unlike control non‐isolated Igs, the antibodies isolated on the Dactylis pollen column were able to inhibit efficiently and in a dose‐dependent manner Dactylis pollen‐induced BHR. The inhibitory activity was increased in isolated antibody samples that had high Ig(i4 levels. Antibodies isolated on the Dactylis pollen column, however, consisted not only of true allergen‐specific (potentially blocking) antibodies but also of autoanti‐Igl‐ binding to allergen‐specific IgE and mistaken for allergen‐specific antibodies, thus opening to question the involvement of the true allergen‐specific antibodies in the BHR‐inhibitory activity. Unlike the true allergen‐specific antibodies, the autoanti‐Igl were retained on and eluted from the anti‐lgF column. Results showed that both the autoanti‐IgE‐depleted and the autoanti‐IgF‐containing fractions accounted for the inhibition observed with the related non‐depleted sample that had been isolated on the Dactylis pollen column.

Conclusion
For the first time, the true blocking activity of allergen‐specific antibodies is demonstrated, that is. in the absence of the autoanti‐Igl which can also inhibit BHR.
Glutathione‐S‐transferase induces murine dermatitis that resembles human atopic dermatitis
Authors: HSU CHING‐HSIANG , CHUA KAW‐YAN , HUANG SHAU‐KU , CHIANGI I‐PING , HSIEH KUE‐HSIUNG
Published: April 2006

Background
The molecular and functional basis of allergen‐induced inflammation seen in atopic dermatitis (AD) remains undefined.

Objective
The objective of this study is to establish a murine model to dissect the pathological mechanisms of inflammatory reactions leading to the development of AD.

Methods
An inbred strain of mice. BALB/c, when injected peritoneally with 30 μg of recombinant Sj26 protein (rSj26). a glutathione S‐transferase of Schistosoma japonicum worm, developed systematic dermatitis 21 days after immunization. The pathology of the dermatitis was examined by histological evaluation and immunostaining. The immediate skin hypersensitivity was demonstrated by scrum transfer and skin test. Epicutaneous patch test was used to demonstrate the antigen‐specific late phase response.

Results
Significant responses of rSj26‐specific IgE were detected 2 weeks after immunization., and such changes paralleled formation of skin lesions. The diseased skin pathology showed inflammatory changes such as infiltration of mononuclear cells and eosinophils in the dermis and mild spongiosis in the epidermis. Numerous IgE bearing cells were also detected in the dermis. Peripheral blood showed eosinophilia at the same time. In addition, rSj26‐specific positive skin test and epicutaneous patch test could be demonstrated in rSj26‐sensitized mice.

Conclusions
These results suggest that rSj26 is capable of eliciting atopic dermatitis‐like lesions in BALB/c mice. This can be a useful animal model for elucidating allergen‐induced immune responses and the development of various therapeutic interventions of atopic dermatitis in humans.
Comparison between hypertonic and isotonic saline‐induced sputum in the evaluation of airway inflammation in subjects with moderate asthma
Authors: BACCI E. , CIANCHETTI S. , PAGGIARO P. L. , CARNEVALI S. , BANCALARI L. , DENTE F. L. , DI FRANCO A. , GIANNINI D. , VAGAGGINI B. , GIUNTINI C.
Published: April 2006

Background
Hypertonic saline‐indueed sputum has recently been used for the evaluation of airway inflammation in asthma.

Objective
To assess the effect of hypertonicity on airway inflammation.

Methods
We compared the inflammatory cell composition of hypertonic saline‐induced sputum with that of isotonic saline‐induced sputum in 21 asthmatic subjects and, at baseline and 30min after each sputum induction, we measured bronchial hyper‐responsiveness to methacholine as an indirect marker to detect increased airway inflammation. On two different days, the patients inhaled hypertonic saline (3–5% NaCl) or isotonic saline (0.9% NaCl) for 30 min via an ultrasonic nebulizer, while monitoring FEV1. Sputum was collected for inflammatory cell analysis.

Results
There was no difference in inflammatory cell percentages obtained with the two methods. Eosinophils were >1% in 20 subjects after hypertonic saline and in 16 subjects after isotonic saline, but this difference was not statistically significant. Intraclass correlation coefficients for sputum inflammatory cells obtained with the two methods were +0.642 for eosinophils, +0.644 for neutrophils. +0.544 for lymphocytes and +0.505 for macrophages. Hypertonic saline induced bronchoconstriction in a significantly greater number of subjects than isotonic saline. Also, hypertonic saline increased bronchial responsiveness to methacholine. while isotonic saline did not.

Conclusion
We conclude that hypertonicity does not affect sputum cell composition, suggesting that inflammatory cells in hypertonic saline‐induced sputum are probably preexisting and not acutely recruited in the airways by the hypertonic stimulus. However, the bronchoconstriction and the increase in bronchial hyper‐responsiveness after hypertonic saline inhalation may imply the release of inflammatory mediators. This fact must be considered in the evaluation of soluble markers of inflammation in hypertonic salineinduced sputum.
Comparison between two automated systems to determine specific IgE: CAP and ELItest
Authors: BRUNNÉE T. , SEEBERGER A. , KLEINE‐TEBBE J. , KUNKEL G.
Published: April 2006

Background and objective
The ELItest® is a newly developed system to measure specific IgE based on allergen bound to paper rings and an alkaline phosphatase conjugated second antibody detection system. It was compared to the CAP® system, a method based on allergen conjugated to an encapsulated cellulose polymer and a β‐galactosidase conjugated fluorescence detection system.

Methods
Sera of 300 patients with positive history and positive skin‐prick tests to common allergens (birch, timothy‐grass, cat dander, dermatophagoides pteronyssinus, wasp venom) and 30 negative controls were tested in both systems. Serial dilutions of high titre sera were measured; inter‐ and intraassay coefficients of variation (cv) were determined.

Results
The CAP system proved to be more sensitive (92.3%) compared to ELItest (84%) but marginally less specific (94.7% for CAP versus 96.7% for ELItest). Intraassay cv were slightly lower in the ELItest (7.2% CAP versus 6.4% ELItest), whereas the interassay cv was roughly twice as high for ELItest (20.1%) than for the CAP system (11.4%). Linearity over an 8‐fold dilution was good in both tests (r2 0.979 ELItest versus 0.996 CAP), although ELItest levelled off at higher allergen concentrations. Similarly, correlation analysis between both systems revealed that ELItest consistently measured lower values, especially at higher concentrations of specific IgE. The slope of the linear regression line of a log/log plot of measured IgE concentrations was significantly lower than 1 in birch, cat and wasp; the y‐intersect was significantly lower than 0 in all analysed allergens.

Conclusion
These results suggest that the ELItest system for the measurement of specific IgE is not quite as reproducible and sensitive as the CAP system but slightly more specific, and that higher concentrations of specific IgE are measured lower in the ELItest. One potential reason might be that the amount of allergen bound to a paper ring might be smaller than that bound to a cellulose polymer, but further experiments are necessary to prove this hypothesis.
Role of sensory neuropeptides in post‐allergic propranolol‐induced bronchoconstriction in guinea pigs in vivo
Authors: FUJIMURA M. , TSUJIURA M. , MYOU S. , ISHIURA Y. , MATSUDA T.
Published: April 2006

Background
Administration of propranolol can provoke bronchoconstriction in asthmatic patients. We hypothesized that such bronchoconstriction may result from the inflammatory mediators released by an allergic reaction. We recently developed a guinea pig model for propranolol‐induced bronchoconstriction (PIB). Neuropeptides which are released from C‐fibre nerve endings have been postulated to induce bronchial hyperresponsiveness through neurogenic inflammation.

Objective
The purpose of this study was to examine whether sensory neuropeptides are involved in the development of PIB after allergic reaction.

Methods
Propranolol at a concentration of 10mg/ml was inhaled 20min after antigen challenge in passively sensitized, anaesthetized and artificially ventilated guinea pigs. The animals were treated intravenously with a NK1 and NK2 dual antagonist, FK224, in a dose of 1 or 10 mg/kg or vehicle or a selective NK1 antagonist, FK888, in a dose of 1 or 10mg/kg or vehicle 10min before or 15min after antigen challenge.

Results
Propranolol inhaled 20 min after antigen challenge caused bronchoconstriction. FK224 or FK888 administered 15 min after antigen challenge as well as 10 min before antigen challenge did not reduce the PIB.

Conclusion
We conclude that neuropeptides such as neurokinin A and substance P do not directly contribute to the development of PIB after allergic reaction.