bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for IgE immune complexes and distribution of IgG subclasses with anti‐IgE activity in patients with atopic dermatitis
Authors: CARINI C. , FRATAZZI C. , AIUTI F.
Published: April 2006

The anti‐IgE autoantibody was detected, using a radioimmunoassay, in 17 out of 35 (48.6%) of patients with atopic dermatitis. Significant increased levels of IgG‐anti‐IgE were seen in the patients studied compared with the control group. The specificity of the anti‐IgE autoantibody was confirmed by competitive inhibition assay using IgG, IgM, IgE myeloma. A correlation was observed between the levels of IgG ‐anti‐IgE and serum IgE but not between the IgG subclasses with anti‐IgE activity and the clinical status. These data demonstrate that the IgG subclass distribution with anti‐IgE activity belongs mostly to the IgG1 and IgG4 subclasses compared with the controls. Moreover, ultracentrifugation analysis indicated that the IgG‐anti‐IgE in the serum samples from the patients with atopic dermatitis was present in the form of an immune complex with self‐IgE. These observations may suggest that the anti‐IgE complexes may play a broader role in the modulation of the immune response and that this autoantibody may mask recognition of IgE in conventional assays.
Tumour necrosis factor stimulates human skin mast cells to release histamine and tryptase
Authors: VAN OVERVELD F. J. , JORENS Ph. G. , RAMPART M. , DE BACKER W. , VERMEIRE P. A.
Published: April 2006

Besides its effects on tumour cells, tumour necrosis factor (TNF) also acts on a variety of other cells, thus enhancing inflammatory and immune processes. In view of the prominent role of the mast cell in such processes, the aim of the present study was to assess the effects of recombinant TNF‐α on human mast cells. Mast cells from the infant foreskin obtained during circumcision were dispersed by an enzymatic technique using collagenasc and hyaluronidase. Cells thus obtained were pooled, washed and separated by Percoll gradient centrifugation. Mast cells, with a purity of 70 90% were incubated for 60 min with 10‐11 to 10‐7MrTNF‐α. Histamine and tryptase levels were assessed in the cell supernatant by spectrofluorometry and radioimmunoassay (RIA) respectively. A concentration dependent release of histamine was observed, which reached a maximum of 11·5 ± 2·2 nmol/106 cells at 10‐8 MrTNF. Release of tryptase was also concentration dependent and reached a maximum of 293±105 mU/106 cells (10‐8 rTNF). rTNF‐α thus appears to be a direct stimulus for mast cells to degranulale and to release both histamine and tryptase.
Genetic ‘risk’ for atopy is associated with delayed postnatal maturation of T‐cell competence
Authors: HOLT P. G. , CLOUGH J. B. , HOLT B. J. , BARON‐HAY M. J. , ROSE A. H. , ROBINSON B. W. S. , THOMAS W. R.
Published: April 2006

Recent in vitro studies suggest that IgE production in adults is co‐ordinately regulated by negative signals from γIFN‐producing CD4+ T‐helper‐1 (TH‐1) and positive signals from IL‐4 producing (TH‐2) T‐cells. Additionally, seroepidemiological evidence has pinpointed infancy as the period of maximum lifetime risk for T‐cell sensitization to ubiquitous environmental antigens. The present study sought to elucidate the relationship between these observations, by examination of CD4+ T‐cell function in normal children and those genetically at‘high risk’ for atopy, spanning the age range (up to 4 years) in which IgE responses to environmental allergens is typically manifest. Immunocompetent T‐cell precursor frequencies (determined by cloning at limiting dilution) were markedly reduced in ‘high risk’ children relative to normals (0.53.0.29 vs 0.26.0.19; P= 0.0025). Consistent with reports from other laboratories employing bulk T‐cell culture techniques, the γIFN producing capacity of CD4+ T‐cell clones from both groups of children were markedly reduced relative to adults, and was lowest in the high risk group (P<0.02). IL‐4 production by CD4+ T‐cell clones from the normal children was within the adult range, but again was significantly lower in the high risk group (P<0.00005). This indicates that initial immune responses to environmental allergens in early childhood occur against a background of maturational ‘deficiency’ in CD4+ T‐cell function, and suggests the possibility that variations in the rate of postnatal maturation of T‐cell competence may be a contributing factor in the development of differing patterns of immunological responsiveness to environmental allergens.
Characterization of a monoclonal antibody (P40) against the 68 kD major allergen of Penicillium notatum
Authors: SHEN H.‐D. , CHOO K.‐B. , CHEN J.‐H. , LIN W.‐L. , CHANG Z.‐N. , HAN S.‐H.
Published: April 2006

A monoclonal antibody (MoAb P40) against the 68 kD major allergen of penicillium notatum (P. notatum) was obtained by immunizing the mouse with a crude extract of P. notatum. Analysed by two‐dimensional gel electrophoresis and immunoblotting, P40 reacted with two different isoforms of the 68 kD component of P. notatum with pIs of 5.4 and 5.5. In addition to P. notatum, P40 showed positive ELISA activity to Aspergillus fumigatus (A. fumigatus) but not to components of six other fungi including Alternaria porri, Cladosporium cladosporoides, Aureobasidium pullulans, Fusarium solani, Rhizopus arrhizus and Candida albicans. Analysed by ELISA, MoAb P40 also showed positive activity to two (P. frequentans and P. roseopurpureum) of the 10 other Penicillium species and two (A. terreus and A. flavus) of the four other Aspergillus species tested. SDS‐PAGE and immunoblotting studies demonstrated P40 positive reactivity to components with MW of about 67 kD in all these Penicillium and Aspergillus species with positive ELISA activity to P40. Furthermore, immunoblotting activity of MoAb P40 to the 67 kD component of A. niger was also observed. The epitope of the 68 kD allergen of P. notatum recognized by MoAb P40 was resistant to treatment of periodate oxidation with concentration of NaIO4 up to 20 mm. This MoAb may thus be useful in the characterization and purification of the 68 kD allergen from crude extracts, and in the molecular cloning of allergen genes.
Monoclonal antibodies bind identically to both spores and hyphae of Aspergillus fumigatus
Authors: REIJULA K. E. , KURUP V. P. , KUMAR A. , FINK J. N.
Published: April 2006

Immunoelectron microscopy (IEM) was used to determine the binding of six monoclonal antibodies (MoAbs) produced against Aspergillus fumigatus antigens present on or within the conidia and hyphae of the fungus. Antigen‐antibody complexes were demonstrated in EM using labelled colloidal gold particles (15 nm). Three out of 6 MoAbs (C9, F12 and H10) reacted only with the cytoplasmic components of A. fumigatus while the remaining three (B12, F6G5 and D6E6) showed reactivity to both cytoplasm and cell wall of the conidia and hyphae. The results indicate that IEM is of considerable value in determining and selecting monoclonal antibodies having specific reactivity with diverse antigenic components.
Comparative efficacy of house dust mite extermination products
Authors: SCHOBER G. , KNIEST F. M. , KORT H. S. M. , DE SAINT GEORGES GRIDELET D. M. O. G. , VAN BRONSWIJK J. E. M. H.
Published: April 2006

The acaricidal efficacy of nine marketed products, i.e. Acardust, Acarosan (foam and powder), Actelic 50, Artilin 3A (spirit and water base), liquid nitrogen, Paragerm AK, and Tymasil, and of intensive vacuum‐cleaning have been compared on four different test surfaces: mattress, tufted carpet, gypsum board and rough wooden board, all covered with artificial house dust. They were inoculated with the house dust mite, Dermatophagoides pteronyssinus or the house‐dust fungus Aspergillus repens for evaluation of the fungistatic claims of some products. The acaricidal activity of Tymasil did not surpass that of vacuuming; its fungistatic effect was not apparent. The other products showed complete to almost complete eradication on at least one of the substrates tested. Taking into account the results of acaricidal efficacy as well as the data on safety and practicality acquired earlier, Acarosan powder was considered first choice for carpet treatment. Acarosan and liquid nitrogen, were found to be effective in the treatment of mattress, pillow, upholstered furniture and heavy curtains. On wooden surfaces Acarosan was found to be both effective and safe, while Acardust, Actelic 50, Artilin 3A (both fungistatic as well as acaricidal), liquid nitrogen and Paragerm also passed the efficiency test.
How allergenic are hypoallergenic infant formulae?
Authors: RUGO E. , WAHL R. , WAHN U.
Published: April 2006

In a comparative study six different protein hydrolysates, marketed as ‘hypoallergenic’ infant formulae were investigated by skin prick tests, RAST, RAST inhibition and titrated provocation tests. Whey hydrolysates containing a high percentage of larger peptides were found to have the highest capacity to induce positive skin tests, provocation tests and. To bind to human serum IgE antibodies of cow's milk allergic children. Casein hydrolysates appeared to have the least residual allergenic activity. We recommend that ‘hypoallergenic’ formulae should be tested in each case, before being prescribed to cow's milk sensitive children.
Egg and milk allergy in adults: comparison between fresh foods and commercial allergen extracts in skin prick test and histamine release from basophils
Authors: NORGAARD A. , SKOV P. S. , BINDSLEV‐JENSEN C.
Published: April 2006

The ability of skin prick test (SPT) and histamine release from basophils (HR) to diagnose clinical type I allergy to egg and milk was investigated as compared with double‐blind, placebo‐controlled food challenge (DBPCFC) in 17 adults suspected of type I egg and/or milk allergy. In both SPT and HR, commercial allergen extracts commonly used for SPT were compared with fresh, standardized foods. With commercial extracts the overall sensitivities of SPT and HR were 0.75 and 0.56 respectively, and none of the tests showed concordance with DBPCFC. With fresh, standardized foods the overall sensitivities of SPT and HR were 1.00 and 0.89 respectively, and both tests now showed a significant concordance with DBPCFC (P<0.05). Specificity was only slightly improved in SPT, and unchanged in HR. Thus, the use of fresh, standardized foods significantly improved the outcome of both tests, as regards to sensitivity and concordance with DBPCFC. The diagnostic ability of SPT and HR appear to be strongly influenced by the allergen quality.
Purification and characterization of major inhalant allergens from soybean hulls
Authors: GONZÁLEZ R. , POLO F. , ZAPATERO L. , CARAVACAJ F. , CARREIRA J.
Published: April 2006

Proteins responsible for respiratory allergy to soybean have been purified from an extract of soybean hulls. The purification procedure combined size exclusion and reverse‐phase HPLC. Two pure glycoproteins (S1 and S2) exhibiting IgE‐binding ability, as demonstrated by immunoblotting and ELISA techniques, were obtained. Both proteins displayed low molecular weight values on SDS‐PAGE (S1, 7.0 kD; S2 7.5 kD). Protein S1 showed charge microheterogeneity, rendering two bands at pH 6.1–6.2 on IEF, whereas S2 showed a single bund at pH 6.8. Amino acid composition analyses revealed a strong homology between S1 and S2 and, as a characteristic feature, a high percentage of hydrophobic residues, mainly leucine and isoleucine. Concerning the allergenic activity, both proteins were recognized by the specific IgE from 95% of patients who suffered asthma attacks during the asthma outbreaks of 1987 and 1988 in Cartagena (Spain), caused by soybean dust. Besides, proteins S1 and S2 were able to, separately, inhibit up to 75% the binding of specific IgE to the whole extract. Moreover, purified proteins totally crossreacted, even though protein S2 seemed to be slightly more active in all the immunochemical techniques employed. Results presented allow us to conclude that both proteins are isoallergens and to name them as Gly m IA (protein S2) and Gly m IB (protein S1), according to the IUIS‐allergen nomenclature system.
Antigen‐specific tachycardia and hypotension in rodents
Authors: LEI H.‐Y. , CHEN H.‐I. , CHAN S.‐H. , LEIR S.‐S. , LIN S.‐B. , WING L.‐Y. CHEN
Published: April 2006

Tachycardia and hypotension, two cardiovascular responses to anaphylaxis, were specifically induced by antigen in mice and rats, respectively, Intravenous injection of poly (Glu60Ala30Tyr10) (GAT) elicited tachycardia within 30–40 sec in GAT‐primed B6 mice. Moreover, a minute amount of GAT (0.2μg) was enough to sensitize the mice to subsequent GAT‐induced tachycardia. Challenging doses ranging from 100 ng to 500 μg could elicit tachycardia. The kinetics of tachycardia induction was different from that of antibody production or delayed‐type hypersensitivity. Tachycardia was induced from day 6 after immunization, while delayed‐type hypersensitivity developed as early as day 4, and anti‐GAT antibodies were undetectable on day 6 and would not reach a maximum until day 8. Specific antigen‐induced hypotension was also observed in rats. Furthermore, cardiovascular changes in both species could be passively transferred by heat‐treated (56°C, 30 min) sera from immunized animals. These benchmarks of antigen‐induced cardiovascular changes in mice or rats could be used as models to study the immune control of cardiovascular changes in anaphylactic responses.
Paracetamol anaphylaxis
Authors: LEUNG R. , PLOMLEY R. , CZARNY D.
Published: April 2006

We describe four cases of anaphylaxis to paracetamol without co‐existing aspirin intolerance and review the Australian experience in paracetamol anaphylaxis. Paracetamol sensitivity may differ in the mechanism from sensitivity to aspirin and other non‐steroidal anti‐inflammatory drugs as previously believed.
Lability in complement receptor mobilization of granulocytes in patients with bronchial hyperreactivity
Authors: LUNDAHL J. , SKEDINGER M. , HED J. , JOHANSSON S. G. O. , ZETTERSTRÖM O.
Published: April 2006

The recruitment of activated granulocytes to bronchial mucosa seems to be involved in the prolonged inflammatory response observed in asthma and probably associated with bronchial hyperresponsiveness. We studied the patients with bronchial hyperresponsiveness with respect to the expression of complement receptor type 1 (CR1), its variability and readiness to become mobilized on peripheral granulocytes from patients with bronchial hyperresponsiveness. CR1 expression and its hourly variation was significantly (P < 0.02, P < 0.01 respectively) higher in the patient group compared with the control group. In addition the ability to mobilize CR1 spontaneously at +37°C correlated to the variation of CR1 expression that occurred during a 4 hr period. These findings indicate that granulocytes from patients with bronchial hyperresponsiveness have a higher degree of variation in CR1 expression that correlates to their ability to mobilize CR1 and may reflect a more pronounced lability.
Stability of Candida albicans allergens during storage
Authors: SAVOLAINEN J. , VIANDER M. , EINARSSON R. , NIEMINEN E. , KOIVIKKO A.
Published: April 2006

Stability of Candida albicans allergens was studied under various storage conditions Lyophilized extract was reconstituted with human serum albumin glycerol‐free and in the presence of 10% or 50% glycerol and stored at various temperatures for different time periods. All extracts were tested at the same time with immunoblotting using C albicans allergic patient sera and galactosidase‐labelled anti‐IgE. The highest number of detected allergens in the immunoblotting pattern was found in the presence of 50% glycerol at +6 C. The most important allergen of $$ the 46 kD protein allergen was stable up to 10 weeks at +6 $$ C in the presence of 50% glycerol but thereafter began to lose its IgE‐binding capacity. After 30 weeks more than 50% of the IgE binding had disappeared. The 27 kD protein, another important allergen, was also labile but retained the allerg$$ better than the 46kD one. The 29kD protein allergen was stable at all storage conditions, except +3$$C tested even after one year. More than 6 months storage at +6C or higher temperature is however unacceptable even in the presence of the 50% glycerol. These findings have particular importance in the diagnosis and treatment of allergic diseases
Presence of IgG4 on the membrane of human basophils. Histamine release is induced by monoclonal antibodies directed against the Fab but not the Fc region of the IgG4 molecule
Authors: JIMENO L. , LOMBARDERO M. , CARREIRA J. , DEL PRADO J. MOSCOSO
Published: April 2006

We have studied the possible role of human IgG4 as an anaphylactic antibody. For that purpose, we have determined the induction of histamine release (HR) from human basophils by anti‐IgE and anti‐IgG4 monoclonal antibodies (MoAbs) recognizing different epitopes located at the Fe and Fab regions of the IgG4 molecule. The results show that anti‐IgG4 (Fab) MoAb was able to induce HR in 93% of donors tested, with no differences between atopics and non‐atopics. That HR is calcium dependent and is accompanied by the synthesis and release of leukotriene C4. In contrast, no HR could be induced by anti‐IgG4(Fc) MoAbs in any individual, even in the presence of D2O or after a second challenge with a polyclonal goat anti‐mouse IgG antibody. The results obtained suggest the presence of IgG4 on the basophil membrane and that the epitope recognized by the anti‐IgG4 (Fc) MoAbs is probably hidden in cell‐bound IgG4. This was demonstrated by immunofluorescence techniques: IgG4 bound to the basophil membrane could be detected with anti‐IgG4(Fab) but not with anti‐IgG4(Fc) MoAbs. In addition, we found that nine donors were unresponsive to an anti‐IgE stimulus, while they released histamine efficiently after challenge with anti‐IgG4(Fab), suggesting the existence of different receptors for both immunoglobulins.
Monoclonal antibodies to Lepidoglyphus destructor: delineation of crossreactivity between storage mites and house dust mites
Authors: HÄRFAST B. , VAN HAGE‐HAMSTEN M. , ANSOTEGUI I.J. , JOHANSSON E. , JEDDI‐TEHRANI M. , JOHANSSON S.G.O.
Published: April 2006

We have developed monoclonal antibodies (MoAbs) to the storage mite Lepidoglyphus destructor (Ld). Employing these anti‐Ld MoAbs (Ld‐MoAbs) in ELISA and ELISA inhibition techniques we have analysed the reaction pattern of Ld‐MoAbs to both non‐pyroglyphid and pyroglyphid mites. The storage mite Glycyphagus domesticus (Gd) exhibited most efficient inhibition, followed by Acarus siro (As), Tyrophagus putrescentiae (Tp), Dermatophagoides pteronyssinus (Dpt) and Euroglyphus maynei (Em). Of the two pyroglyphid species, Dpt showed at least 1000 times less inhibition than Gd. Two of the MoAbs immunoprecipitated a band of 39 kD whereas the third reacted weakly, with a high‐molecular band of approximately 110 kD. The Ld extract was also subjected to various reagents and conditions and the antigen was heat stable, it was not affected by low pH, or sensitive to dimethyl sulphoxide (DMSO) or paraformaldehyde. After exposure of the extract to various reagents, such as protease trypsin and periodate, we conclude that the epitopes recognized by Ld‐MoAbs were of carbohydrate rather than of protein nature. It would thus seem that MoAbs recognize the carbohydrate part of a glycoprotein.
House dust mite sensitivity: interaction of genetics and allergen dosage
Authors: YOUNG R. P. , HART B. J. , MERRETT T. G. , READ A. F. , HOPKIN J. M.
Published: April 2006

Pairwise analysis of siblings from 21 families showed that house dust mite (HDM) sensitive children were exposed to higher concentrations of Der p I allergen in their mattress (P= 0.005) and bedding (P= 0.04), but not bedroom floor (P= 0.33), than their atopic sibling who was not sensitive to HDM antigens. There was no difference in the exposure to HDM numbers/100 mg of dust in the mattress (P= 0.61) or bedroom floors (P= 0.09). In contrast, pairwise analysis of siblings from 15 families showed that HDM sensitive children were not exposed to significantly different concentrations of Der p I in the mattress (P= 0.96), bedding (P= 0.11) or bedroom floor (P= 0.70) nor HDM numbers/100 mg of dust in the mattress (P= 0.12) and bedroom floor (P= 0.98) than their non‐atopic siblings. These findings were identical when absolute allergen load was compared in these pairs. Genetic linkage studies in these families suggest the tendency to atopic IgE responses is conferred by a putative atopy locus on chromosome 11q. These results together suggest that differences in allergen levels in beds, among siblings with a comparable genetic tendency to atopy, play a significant role in determining the development of HDM allergy.
The effect of azelastine on the early allergic response
Authors: SHIN M.‐H. , BAROODY F. , PROUD D. , KAGEY‐SOBOTKA A. , LICHTENSTEIN L. M. , NACLERIO R. M.
Published: April 2006

To study the effect of azelastine on the immediate reaction to nasal allergen challenge, we performed a double blind, placebo‐controlled cross‐over clinical trial. Thirteen subjects with seasonal allergic rhinitis underwent nasal challenge with antigen 4 hr after a single oral 2 mg dose of azelastine. The response was monitored by counting the number of sneezes and by measuring the levels of histamine, prostaglandin D2, immunoreactive sulphidopeptide leukotrienes, kinins and TAME‐esterase activity in recovered nasal lavages. After a single dose of azelastine, there was a significant reduction in sneezing (10 vs 2, P= 0.01) and in the median levels of recovered TAME‐esterase activity (63.1 vs 17.5 c.p.m. × 10‐3, P= 0.01), immunoreactive sulphidopeptide leukotrienes (7.5 vs 2.1 ng/ml, P= 0.03) and kinins (1370 vs 251 pg/ml, P= 0.03), with no significant reduction in the median levels of histamine (3.7 vs 1.2 ng/ml, P= 0.2) and prostaglandin D2 (70 vs 70 pg/ml, P= 0.2) compared to placebo (numbers represent total increase over diluent challenge). These results suggest that azelastine does not inhibit mast cell activation but affects the consequences of released histamine, namely sneezing, increased vascular permeability and the generation of kinins. The results further suggest that other cells, in addition to mast cells, might be responsible for the generation of leukotrienes during the early allergic response, and that azelastine reduces their ability to generate this mediator or that inhibition of leukotriene release from mast cells occurs at lower drug concentrations.
Comparison of mechanisms of IL‐3 induced histamine release and IL‐3 priming effect on human basophils
Authors: OKAYAMA Y. , BEGISHVILI T. B. , CHURCH M. K.
Published: April 2006

We have investigated the mechanisms by which inlerleukin‐3 (IL‐3) induces histamine release and primes basophils for increased hislamine release in response to anti‐IgE‐ and N‐formyl‐methionyl‐leucyl‐phenylalanine (fMLP). The responsiveness of basophils from atopic donors was variable, only 5/11 subjects showing release of > 10% to IL‐3 in the range 0.1–100 ng/ml. IL‐3‐induced histamine release required both extracellular Ca2+ and cell membrane IgE, removal of membrane IgE by lactate stripping or desensitization of basophils by incubation with anti‐IgE in a Ca2+‐free medium blocking IL‐3‐induced hislamine release.

IL‐3 also primed basophils for histamine release by anti‐IgE and fMLP in the same concentration range as it evoked histamine release. When IL‐3 and either anti‐IgE or fMLP were combined, the result was additive or supra‐additive depending on the basophil donor. Unlike IL‐3‐evoked histamine release, IL‐3 priming of basophils for fMLP‐induced histamine release was shown to be independent of the presence of both cell surface IgE and of extracellular calcium. The protein kinase C (PKC) inhibitor, staurosporine (10 nM), inhibited anti‐IgE induced histamine release, but neither IL‐3 induced histamine release nor IL‐3 priming of IgE‐ and fMLP‐induced histamine release. Pertussis toxin (1.0 μg/ml) inhibited fMLP‐induced histamine release but not anti‐IgE‐induced histamine release, IL‐3‐evoked histamine release or IL‐3 priming. These results indicate that IL‐3 modulates mediator release from human basophils by two mechanisms; a direct release of histamine which involves cell surface IgE and the influx of extracellular calcium but which is unlikely to proceed by the same mechanism as cross‐linkage of IgE, and a priming effect which is independent of IgE and extracellular Ca2+ and which enhances the secretory effects of a wide range of unrelated secretagogues.
Expression of CD35 (CR1) and CD11b (CR3) on circulating neutrophils and eosinophils from allergic asthmatic children
Authors: BERENDS C. , HOEKSTRA M. O. , DIJKHUIZEN B. , DE MONCHY J. G. R. , GERRITSEN J. , KAUFFMAN H. F.
Published: April 2006

Complement receptors on neutrophils and eosinophils play a role in activation and adhesion. During asthmatic reactions these receptors have been found elevated on circulating granulocytes. In the present study we compared the expression of CD35 (complement receptor type 1) and CD11b (complement receptor type 3) on neulrophils and eosinophils from asthmatic and non‐asthmatic children. This was done in whole blood samples using depolarised light scattering for the discrimination of neutrophils and eosinophils. The non‐stimulated expression as well as the upregulaled expression of receptors by the chemotactic peptide N‐formylmethionyl‐teucyl‐phenylalanine (fMLP) were studied. The results showed that without prior stimulation only the expression of CD35 on neutrophils was significantly elevated in children with asihma (P < 0.05). After up‐regulation with fMLP. the CD11b expression on neutrophils (P < 0.005, fMLP: 0.002 μm) and eosinophils (P < 0.05, fMLP: 0.02 μM) was significantly higher in asthmatic children than in the controls. These results indicate that the inducible expression of CD11b on neutrophils and eosinophils from allergic asthmatic children is primed in rivo.
Spontaneous histamine release in whole blood in patients before and after 4 months of specific immunotherapy
Authors: WANTKE F. , GÖTZ M. , JARISCH R.
Published: April 2006

Spontaneous histamine release (SHR) in whole blood was assessed before and after 4 months of specific immunotherapy (SIT) for allergic rhinoconjunctivitis in 32 patients. Spontaneous histamine release was significantly enhanced (P < 0.05) in patients prior to immunotherapy compared with 20 controls. Spontaneous histamine release decreased significantly in patients after 4 months of specific immunotherapy (P < 0.04) and almost reached the same values as spontaneous histamine release in controls. Clinical success of treatment after 4 months was seen in 15 patients (improvement 50%), 10 of whom showed a significant decrease in spontaneous histamine release. Decrease of spontaneous histamine release after 4 months indicates the efficacy of specific immunotherapy already at an early stage of treatment. Assessment of spontaneous histamine release appears to be a useful and easily performable method for monitoring success of treatment of patients during specific immunotherapy.
Pistachio nut hypersensitivity: identification of pistachio nut allergens
Authors: PARRA F. M. , CUEVAS M. , LEZAUN A. , ALONSO M. D. , BERISTAIN A. M. , LOSADA E.
Published: April 2006

Type I hypersensitivity to pistachio nut antigens was demonstrated in three patients by means of immediate skin‐test reuctivily. specific IgF determination by a lluoro‐immunoassay (CAP). CAP‐inhibition and leucocyte histamine release. Sensitization to other dried fruils and pollens was observed in the patients. The CAP‐inhibition studies revealed signilicLint crossreactivity between pistachio and cashew nut belonging to the Anacardiaceae family, and between pistachio nut and other dried fruits belonging to taxonomically unrelated botanical families. No relevant crossallergenicity was observed between pistachio nut and Lolium and Olca pollens.

Sodium dodecyi sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) of a pistachio nut extract followed by immunoblotting analysis identified four IgE‐binding bands with molecular weights of 34. 41. 52 and 60 kD.
Effects of the platelet activating factor antagonists BN 52021 and BN 50730 on antigen‐induced bronchial hyperresponsiveness and eosinophil infiltration in lung from sensitized guinea‐pigs
Authors: LAGENTE V. , BOICHOT E. , CARRE C. , GUINOT PH. , MENCIA‐HUERTA J. M. , BRAQUET P.
Published: April 2006

The involvement of platelet activating factor (PAF) in antigen‐induced bronchial hyperresponsiveness was investigated by the use of the PAF antagonists BN 52021 and BN 50730, in a guinea‐pig model where sensilization and challenge were performed by aerosol. Male Hartley guinea‐pigs were sensitized by two aerosol exposures at 48 hr intervals to a 0.9% NaCl solution (saline) containing 2 mg/ml ovalbumin lor 30 min. Fifteen to 20 days later, guinea‐pigs were challenged by exposure to five successive aerosols ofincreasing concentrations of ovalbumin (OA) or respectively, 10 μg ml. KM) μ/ml, 1 mg/ml. 5 mg/ml and 10 mg/ml for 15 min each, orsaline alone. Three to four hr and 18‐24 hr after the aerosol challenge the guinea‐pigs were prepared for recording of bronchopulmonary response and aerosol administrations were then generated with an ultrasonic nebulizer. The bronchopulmonary responses induced by successive I‐min aerosol bursts of acetylcholine(ACh) was assessed. As compared with saline‐challenged guinea‐pigs, an enhanced bronchopulmonary response to aerosol administration of cumulative doses of ACh was observed, 3 4 hr and 18 24 hr post‐ovalbumin challenge. When the sensitized guinea‐pigs were pretreated I hr before ovalbumin exposure with BN 52021 or BN 50730 (25 mg/kg, per ox), a significant inhibition of the increase in the bronchopulmonary response to ACh was observed, both at 3–4 hr and 18–24 hr. Furthermore, when guinea‐pigs were treated 3–4 hr after the ovalbumin exposure with BN 52021 or BN 50730, a significant inhibition of the hyperresponsiveness to ACh was recorded at 18 24 hr. A marked accumulation of eosinophils in the peribronchial regions was observed on histological preparations of lung specimens collected 4 hr or 24 hrafler ovalbumin exposure. Pretreatment of the guinea‐pigs by BN 50730 or BN 52021 did not modify the eosinophil accumulation in the peribronchial area. No significant difference in the number of eosinophils collected in the bronchoalveolar lavage fluid is observed, 24 hr posl‐ovalbumin challenge, under the pretreatment with BN 52021 or BN 50730. Pretrealment of guinea‐pigs by BN 50730 or BN 52021 significantly reduced the PAF‐induced (100μg/ml) increase in eosinophil number in the peribronchial area. By contrast, they did not inhibit the eosinophilia induced by aerosol administration of LTB4 (5 μg/ml). These results suggest that the bronchial hyperresponsiveness observed in this study is associated with eosinophil accumulation in the lung. The potent inhibition of the bronchial hyperresponsiveness by the two unrelated antagonists of PAF suggests that the lipid mediator is involved in its triggering and duration, but not in the eosinophil infiltration.
Neutrophil–induced human bronchial hyper responsiveness in vitro—pharmacological modulation
Authors: HUGHES J. M. , McKAY K. O. , JOHNSON P. R. , TRAGOULIAS S. , BLACK J. L. , ARMOUR C. L.
Published: April 2006

Although it has been postulated that inflammatory cells cause the bronchial hyper‐responsiveness which is diagnostic of asthma [1], until recently there has been little direct evidence of such a link. We have recently shown that calcium ionophore‐activated human neutrophils and eosinophils can induce a state of human airway hyperresponsiveness in vitro [2]. In this study we have shown that the anti‐inflammatory agent nedocromil sodium, 10‐7m, inhibited the hyperresponsiveness induced by products released from ionophore activated neutrophils but did not inhibit the release of leukotriene B4 from the same cells. Neutrophil‐induced bronchial hyperresponsiveness was also inhibited by pre‐treatment of the bronchial tissues with a thromboxane A2 and prostaglandin receptor antagonist, GR32191,10‐7m. These findings indicate that cyclooxygenase products are involved in bronchial hyperresponsiveness induced by inflammatory cell products in vitro and that their release can be inhibited by nedocromil sodium.
Lack of increased numbers of low‐density eosinophils in the circulation of asthmatic individuals
Authors: BRUIJNZEEL P. L. B. , JNR. J‐CHR VIRCHOW , RIHS S. , WALKER C. , VERHAGEN J.
Published: April 2006

The density distribution pattern of eosinophils over discontinuous isotonic Percoll gradients from the blood of normal, asymptomatic allergic and non‐allergic asthmatic individuals was investigated. There was a completely identical distribution pattern between the investigated groups. Analysis of the expression of surface markers for complement receptors CR1 and CR3 and immunoglobulin G receptor on eosinophils derived from the density bands 1.080. 1.085 and 1.090 g/ml supported this finding since they did not reveal differences in expression between the bands within one group but also not between the three groups. Eosinophils of the various density bands were further purified and stimulated in vitro to produce leukotriene C4 (LTC4) by the calcium ionophore A23187 or serum treated zymosan. Equal amounts of LTC4 were synthesized by the eosinophils of the various density bands within one group. However, it appeared that the eosinophils of all density bands of allergic and non‐allergic asthmatics synthesized significantly more LTC4 than the eosinophils from normal individuals (five‐to tenfold). Probably this indicates in vivo priming of the eosinophils in asthmatic individuals which is not reflected by a change in density. Control experiments, dealing with possible artifacts due to the isolation procedure or the patient selection, to find differences in distribution patterns over discontinuous Percoll density gradients of the eosinophils of asthmatic compared to normal individuals failed to show such a difference. Therefore, the density distribution pattern of eosinophils over these gradients does not reflect cell activation, whereas LTC4 formation clearly does. This could mean that LTC4 formation is a more sensitive parameter for cell activation than density distribution or cell surface marker expression.
Increase in non‐specific bronchial responsiveness after repeated inhalation of low doses of allergen
Authors: IHRE E. , ZETTERSTROM O.
Published: April 2006

The development of bronchial hyperresponsiveness (BHR) in asthma is considered to be caused by inflammation of the airway. In IgE‐mediated allergy BHR is related to the occurrence of late phase reactions. We have previously shown that exposure to low doses of allergen can cause isolated late reactions. These findings are potentially of clinical importance, since exposure to low, subclinical allergen doses may lead to bronchial inflammation and increasing bronchial responsiveness without necessarily causing immediate bronchoconstriction. This study was performed to investigate whether repeated exposure to low doses of allergen could induce a change in BHR. The trial comprised two groups of five and eight patients with a history of allergic asthma. They were submitted to a series of allergen inhalations for 5–7 days. They were given the same low allergen dose (1–10 biological units) each day. Before and after the allergen exposure period histamine challenges were performed. After the week of allergen inhalation the bronchial responsiveness was increased in 11 of 13 patients.
The main allergen of Olea europaea (Ole e I) is also present in other species of the oleaceae family
Authors: OBISPO T. M. , MELERO J. A. , CARPIZO J. A. , ARREIRA J. C. , LOMBARDERO M.
Published: April 2006

Three major pollen allergens from Fraxinus excelsior, Ligustrurn vulgare and Syringa vulgaris belonging to the Oleaceae family were purified. Monoclonal antibodies previously raised against the main allergen of Olea europaea (Ole e I) were used for their purification by affinity chromatography. The three new purified allergens were able to bind human IgE from serum of olive‐allergic patients in a way analogous to Ole e I. Crossed radioimmunoelectrophoresis of the four allergens, using anti‐olive extract rabbit serum, showed a unique immunoprecipitation arc with the same characteristics. The four purified proteins had similar molecular weights on SIX‐PAGE and the N‐terminal sequences for the first 20 amino acids were identical. Furthermore, the concentration of the allergens could be determined using a two‐site solid phase assay previously developed for the allergen Ole e I. Our results indicate that the four purified proteins share, to a great extent, antigenic and allergenic epitopes leading to cross‐reactivities which could cause common clinical manifestations. We propose for the newly purified allergens the nomenclature of Fra e I, Lig v I and Syr v I.
Steroid sparing effect of nedocromil sodium in asthmatic patients on high doses of inhaled steroids
Authors: WONG C. S. , COOPER S. , BRITTON J. R. , TATTERSFIELD A. E.
Published: April 2006

Nedocromil sodium is a non‐steroidal prophylactic agent developed for the management of asthma. We have assessed the steroid sparing potential of inhaled nedocromil sodium 4 mg four limes daily in a randomized, double blind, placebo controlled study in 69 asthmatic subjects controlled on inhaled beclomethasone dipropionate in the dose range 1000 2000 μg daily. Following a 4 week run‐in period subjects added nedocromil sodium or placebo by metered dose inhaler to their usual medication for a further 4 weeks. The dose of inhaled steroid was then reduced at fortnightly intervals according to a predetermined schedule, with monitoring of asthma severity, symptom scores, bronchodilator use and peak flow recordings. Sixty subjects entered the steroid reduction phase and achieved median (range) % decreases in steroid dose of 80 (17‐100)% with nedocromil sodium compared to 65 (0‐100)% with placebo (P = 0.34) with 14 patients in the nedocromil sodium group and 10 in the placebo group being withdrawn completely from inhaled steroids. Subjective global assessment scores were significantly better with nedocromil sodium (mean 2.14) than with placebo (2.93; P<0.02) though there was no difference between individual daily symptom scores. In this study therefore in asthmatic patients controlled on high doses of inhaled steroids, nedocromil sodium was well tolerated but the smalt differences in steroid sparing effect between nedocromil and placebo were not statistically significant.
Monoclonal antibodies specific for guinea pig eosinophil major basic protein: their use in an ELISA, immunocytochemistry and flow cytometry
Authors: HUNT T. C. , SUMMERS J. A. , CAMPOS M. G. , RIMMER S. J. , STURTON G. , PALFAI S. , CHURCH M. K.
Published: April 2006

Eosinophil, major basic protein (M BP), purified from guinea pig eosinophil granules was used to raise five monoclonal antibodies (MoAbs). Their reactivity with MBP was confirmed by immunoblotting and indirect ELISA. Two of the MoAbs were used to develop a sensitive and specific antigen capture (sandwich) ELISA for guinea pig eosinophil M BP which gives an accurate and reproducible standard curve over the range of 10‐10000 ng/ml. The specificity of the ELISA for MBP was confirmed and its suitability for testing biological samples ascertained by measurement of MBP in bronchoalveolar lavage fluid (BALF) and plasma from guinea pigs sensitized and challenged with ovalbumin. The ELISA was also capable of detecting MBP in culture supernatants from purified eosinophil preparations challenged with calcium ionophore in vitro. One of the monoclonal could be used to strongly and specifically stain guinea pig eosinophils in immunocytochemistry, whilst all five could be used to visualize eosinophils in suspension in BALF or peritoneal lavage fluid by flow cytometry. There was no staining of other guinea pig leucocyte types, nor crossreactivity with human eosinophils by immunocytochemistry or Row cytometry.
Exposure to house dust mite allergen of children admitted to hospital with asthma
Authors: SPORIK R. , PLATTS‐MILLS T. A. E. , COGSWELL J. J.
Published: April 2006

Eighty‐two children admitted to hospital with exacerbations of asthma were studied to determine how many were exposed to house dust mites at the time of admission and displayed immediate hypersensitivity to house dust mites. The concentration of house dust mite allergen (Der p I) was measured in dust obtained from the child's mattress, bedroom floor and living room floor. Sixty‐two (75%) children admitted had been exposed to > 10 μg Der p I/g. Sixty‐seven (82%) children were sensitive to house dust mite(RAST 1 +, or weal 3 mm): 49 (60%) children were both exposed and sensitive. In contrast in a control group of 44 children, 31 (70%)(n.s.) were exposed to > 10μg Der p I/g, 10 (23%) (P < 0.001) were sensitive to house dust mite, and 7 (16%) (P < 0.001) were both exposed and sensitive. Seventy‐three homes were revisited 6 months after the child's initial admission. During the preceding month 14 children had been readmitted, 12 were fully investigated; of these 10 were both sensitive to house dust mite and still exposed to > 10 μg Der p I/g. In contrast, of the remaining 62 children who were not readmitted, only 19 were both sensitive and still exposed to > 10μg Der p I/g (P < 0.001). In conclusion, the majority of children admitted to hospital with exacerbations of asthma were exposed to house dust mite allergen and were house dust mite sensitive. Further the results suggest that continued exposure to higher concentrations of mite allergen may be associated with the risk of readmission.
The renin angiotensin system and hymenoptera venom anaphylaxis
Authors: HERMANN K. , RING J.
Published: April 2006

Components of the renin angiotensin system, namely renin, angiotensinogen, angiotensin I and II and aldosterone were measured in plasma of patients with hymenoptera venom anaphylaxis (n= 50) and healthy non‐allergic controls (n= 25). Patients with a history of anaphylactic reactions to hymenoptera venom who did not undergo immunotherapy showed significantly reduced renin, angiotensinogen, angiotensin I and angiotensin II in plasma as compared with controls (P<0.05). There was no difference in the aldosterone concentration between patients and controls. Angiotensin I, angiotensin II, renin and angiotensinogen levels were the same in male and female patients. There was also no difference in the angiotensin I, II, renin or angiotensinogen levels between young and older patients. A significant inverse correlation between the severity of clinical symptoms and the plasma levels of renin (r=‐0.382, P<0.001), angiotensinogen (r=‐0.567, P<0.0001), angiotensin I (r= ‐0.656, P<0.0001) and angiotensin II (r = 0.0762, P < 0.0001) was found: the lower the levels the more severe the clinical symptoms. No correlation was found for aldosterone. Hymenoptera venom allergic patients with repeated anaphylactic reactions during hyposensitization did not tolerate the sting of a living insect (n= 6). In these patients, renin, angiotensinogen, angiotensin I and II remained significantly lower than in healthy non‐allergic controls. Patients with successful immunotherapy (n= 27) who tolerated the sting of a living insect had renin, angiotensin I and II significantly higher than patients without immunotherapy. These findings suggest a possible role of the renin angiotensin system in hymenoptera venom anaphylaxis.
Treatment of steroid‐dependent asthma with recombinant interferon‐gamma
Authors: BOGUNIEWICZ M. , SCHNEIDER L. C. , MLLGROM H. , NEWELL D. , KELLY N. , TAM P. , IZU A. E. , JAFFE H. S. , BUCALO L. R. , LEUNG D. Y. M.
Published: April 2006

We have recently reported that treatment of patients with severe atopic dermatitis with recombinanl interferon‐gamma (rIFN‐y) resulted in clinical improvement as well as a reduction of circulating eosinophils. Since IgE‐dependent late phase allergic reactions and eosinophilic infiltration are thought to play an important role in the pathogenesis of asthma, we conducted a two centre randomized double‐blind placebo‐controlled trial of rIFN‐y in the treatment of steroid‐dependent asthma. Patients were treated with daily subcutaneous injections of either 0.05 mg/m2 rIFN‐y (n= 9) or placebo (n=11) for 90 days. All patients completed the study without significant drug toxicity noted. Oral prednisone dose, forced expiratory volume in 1 second (FEV), peak expiratory flow rates (PEFR) and circulating eosinophil counts were monitored throughout the trial. There was no significant difference between the two treatment groups in per cent reduction from baseline of daily prednisone (P=0.51). There was also no significant difference between the two treatment groups in per cent change from baseline in FEV1 (P= 0.54)or in PEFR (P=0.75). Total circulating eosinophil counts decreased by 31% in the rIFN‐y group and increased by 8.5% in the placebo group (P= 0.09). We conclude that this treatment regimen was not effective in patients with steroid‐dependent asthma.
Occupational allergy to bumble bee venom
Authors: KOCHUYT A. M. , VAN HOEYVELD E. , STEVENS E. A. M.
Published: April 2006

The clinical profile of anaphylactic reactions to bumble bees is described and successful immunotherapy with honey bee venom in seven bumble bee allergic patients is reported. The cause of the high frequency of sensitization to pollen in these patients is discussed.
In vitro allergenicity of cows' milk substitutes
Authors: DEAN T. P. , ADLER B. R. , RUGE F. , WARNER J. O.
Published: April 2006

There are numerous alternatives to cows' milk formula for allergic children. We have investigated the allergenicity of several of these using RAST and RAST inhibition on serum from 16 patients with a known history of cows' milk protein intolerance (CMPI) and 16 atopic controls. A RAST grade of ≥3 for cows' milk was present in all those with CMPI, whilst all the controls gave RAST of ≤1 ‐ Modified cows' milk formula, goats' infant formula, sheep and goats' milk produced similar results to cows' milk. Only two patients had RAST ≥3 for soya milk and the soy/beef hydrolysate gave positive results in only three patients. One had positive RAST to Nutramigen and two to Pregestimil. Of the whey hydrolysates investigated, Pepti‐junior gave seven positive RASTs whilst we were unable to hind Alfare to the sepharose in sufficient quantities to interpret the results which were negative in all cases. RAST inhibition data on pooled sera from the same patients agreed with the RAST results. The inhibition curves showed high inhibition with goats', sheep, modified cows' milk formula and the casein formula. AL110 (50%). Soy and soy/beef hydrolysate showed a much lower inhibition pattern. Casein hydrolysates showed low inhibition while the whey hydrolysate produced higher inhibition. We have shown that despite claims of low allergenicity. some of these alternative formulae are antigenically recognized in vitro by some cows' milk intolerant patients.