bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of isoform‐specific T‐cell epitopes in the major timothy grass pollen allergen, Phl p 5
Authors: Würtzen P. A. , Bufe A. , Wissenbach M. , Madsen H. O. , Ipsen H. , Arnved J. , Van Neerven R. J. J. 
Published: March 2003

BackgroundThe involvement of CD4+ T cells in the pathophysiology of atopic disease is well established.ObjectiveTo gain further insight into the activation requirements for allergen‐specific T cells, we characterized epitope specificity, HLA restriction and T‐cell receptor (TCR) usage for T cells specific to Phl p 5, the group 5 major allergen of the grass Phleum pratense.MethodsTo identify the T‐cell epitopes of Phl p 5, three Phl p 5‐specific T‐cell lines (TCLs) and 15 T‐cell clones (TCCs) generated from the peripheral blood of three grass‐allergic patients were tested with recombinant truncated Phl p 5a fragments and synthetic Phl p 5b peptides representing these two different recombinant Phl p 5 isoallergens. Additional activation experiments with HLA‐subtyped antigen‐presenting cells and flow cytometry analysis with TCR V‐specific mAb were performed to further characterize the activation requirements for Phl p 5‐specific TCCs.ResultsAt least nine distinct T‐cell specificities were identified and the T‐cell epitopes recognized differed considerably among the three patients. Most of the epitopes found were isoform‐specific, whereas three epitopes were shared between Phl p 5a and 5b. Several human leucocyte antigen (HLA) class II molecules were involved in the recognition of Phl p 5. Different HLA restriction specificities were even found among TCCs specific to the same epitope region. All TCCs were TCR‐α/β positive, and an overrepresentation of TCR Vβ 3.1+ clones among TCCs specific to Phl p 5 appear to exists as 31% (4/13) of the TCCs expressed TCR Vβ 3.1 (compared with 5% TCR Vβ 3.1+ T cells in human peripheral blood) with no correlation with epitope specificity or HLA restriction.ConclusionThe T‐cell reactivity of the three grass‐allergic patients investigated shows that isoallergen‐specific T‐cell epitopes are found throughout the peptide backbone of Phl p 5a and Phl p 5b, and dominant T‐cell epitopes of Phl p 5 were not identified. This indicates that a mixture of at least full‐length rPhl p 5a and rPhl p 5b may be required to target the total Phl p 5‐specific T‐cell response of atopic patients.
Exposure to systemic prednisolone for 4 hours reduces ex vivo synthesis of GM‐CSF by bronchoalveolar lavage cells and blood mononuclear cells of mild allergic asthmatics
Authors: Cotter T. P. , Hood P. P. , Costello J. F. , Sampson A. P. 
Published: March 2003

BackgroundIn acute severe asthma, the earliest clinical effects of glucocorticosteroids occur from 4 to 5 h after systemic administration, but the mechanisms are unclear. In persistent asthma, corticosteroids are thought to suppress airway inflammation by modulating the expression of adhesion molecules, enzymes, and leucotactic cytokines, including granulocyte‐macrophage colony stimulating factor (GM‐CSF). GM‐CSF is also overexpressed in the airways of symptomatic asthmatics.ObjectivesTo examine the early effects of systemic corticosteroids on cytokine expression, we investigated whether ex vivo synthesis of GM‐CSF is suppressed in the bronchoalveolar lavage (BAL) cells and peripheral blood mononuclear cells (PBMCs) of normal and mild allergic asthmatic subjects obtained 4 h after a single intravenous dose of prednisolone.MethodsIn a randomized, double‐blind, placebo‐controlled study, BAL cells and PBMCs were obtained from mild atopic asthmatic patients (n = 9) and normal subjects (n = 9) 4 h after an intravenous bolus dose of 80 mg prednisolone, and cultured for 0–18 h in the presence or absence of lipopolysaccharide (LPS; 10 μg/mL). Enzyme immunoassay was used to assess GM‐CSF levels in BAL cell and PBMC culture supernatants, and in BAL fluid.ResultsAfter placebo, GM‐CSF synthesis tended to be higher in BAL cells from asthmatics than in normals. LPS stimulation significantly increased median (interquartile range) GM‐CSF synthesis by BAL cells ex vivo from 16.4 (23 to 74) to 35.8 (3–148) pg/106 cells in normals (P < 0.05), and from 59 (9 to 204) to 134 (24–288) pg/106 cells in asthmatics (P < 0.01). After intravenous prednisolone, the rise in GM‐CSF production induced in BAL cells by LPS was completely abolished in both subject groups. In PBMCs of placebo‐treated asthmatics (but not normals), LPS stimulated median GM‐CSF synthesis from 164 (110 to 300) to 314 (235–485) pg/106 cells (P = 0.02), and this was blocked by intravenous prednisolone.ConclusionsLPS‐stimulated GM‐CSF synthesis ex vivo is abolished in BAL cells of mild asthmatic and normal subjects, and in PBMCs of asthmatics, obtained 4 h after a single intravenous dose of prednisolone. Suppression of GM‐CSF synthesis in airway and blood leucocytes may contribute to the early clinical efficacy of systemic glucocorticoids in acute allergic asthma.
The use of phage‐peptide libraries to define the epitope specificity of a mouse monoclonal anti‐Der p 1 antibody representative of a major component of the human immunoglobulin E anti‐Der p 1 response
Authors: Furmonaviciene  , Tighe  , Clark  , Sewell  , Shakib  
Published: March 2003

BackgroundMore than 80% of individuals who are sensitive to the dust mite Dermatophagoides pteronyssinus produce immunoglobulin (Ig) E antibodies to Der p 1, the most significant domestic allergen. There is therefore considerable interest in elucidating the interaction between human IgE and Der p 1, as a basis for developing strategies for therapeutic intervention.ObjectivesWe have therefore sought to determine the Der p 1 epitope recognized by a mouse monoclonal anti‐Der p 1 antibody (mAb 2C7) representative of a major component of the human IgE anti‐Der p 1 response.MethodsM13 15mer and T7 9mer bacteriophage‐peptide display libraries were screened with mAb 2C7. Mimotope sequences were defined and compared with the native Der p 1 sequence and with those of three homologous molecules, namely chymopapain, papain and actinidin. The sequence of a candidate epitope was then located in the three‐dimensional model of Der p 1 and the corresponding sequences in the homologous molecules were studied for accessibility in the three‐dimensional structure.ResultsWe have demonstrated that it is possible to isolate phage clones with peptide inserts specific for mAb 2C7. Examination of the sequences obtained and the location of the corresponding epitope within the three‐dimensional model of Der p 1 has shown that mAb 2C7 recognizes a conformational epitope comprising the sequence Leu147‐Gln160. The relevance of the identified epitope was established by showing that native Der p 1 can block the binding of specific phage clones to mAb 2C7. Similar sequences were identified within the three‐dimensional structures of chymopapain, papain and actinidin, thereby providing a structure‐based explanation for immunological cross‐reactivity.ConclusionThe identification of the Der p 1 sequence Leu147‐Gln160 as a potential epitope recognized by a major component of the human IgE anti‐Der p 1 response may provide therapeutic opportunities for disrupting the interaction between IgE and this important allergen.
Airway responsiveness to acetaldehyde in patients with asthma: relationship to methacholine responsiveness and peak expiratory flow variation
Authors: Prieto  , Sánchez‐Toril  , Brotons  , Soriano  , Casañ  , Belenguer  
Published: March 2003

BackgroundAlthough airway hyperresponsiveness to inhaled acetaldehyde has been documented in Japanese patients with asthma, the response to this bronchoconstrictor agent has never been studied in Caucasians.ObjectivesThe objectives of the study were to determine differences in airway responsiveness to acetaldehyde between asthmatic and healthy subjects, and to examine the relationship between acetaldehyde responsiveness and the variability of peak expiratory flow (PEF).MethodsThe response to methacholine and acetaldehyde challenges was measured in 81 non‐smoking adults (61 asthmatics and 20 normal controls). Subjects recorded PEF morning and evening for 14 days. The response to both bronchoconstrictor agents was measured by the PC20 (provocative concentration required to produce a 20% fall in FEV1). PEF variation was expressed as amplitude percentage mean, and as low percentage best (lowest PEF expressed as a percentage of the best PEF recorded).ResultsThe two types of challenge yielded a similarly high level of sensitivity (100% for methacholine and 92% for acetaldehyde) and specificity (90 and 100%, respectively) to distinguish between asthma and controls. Asthmatic subjects were on average 265‐fold less sensitive to acetaldehyde than to methacholine. PC20 acetaldehyde correlated weakly but significantly with both indices of PEF variation (amplitude percentage mean: ρ = − 0.36, P = 0.004; low percentage best: ρ = 0.42, P = 0.001).ConclusionsThese results indicate that airway hyperresponsiveness to acetaldehyde is a sensitive and specific indicator for separating asthmatic and normal subjects. Airway responsiveness to methacholine or acetaldehyde and PEF variation are not reflecting the same pathophysiological process in the airways.
Dose‐related efficacy and tolerability of fluticasone propionate nasal drops 400 μg once daily and twice daily in the treatment of bilateral nasal polyposis: a placebo‐controlled randomized study in adult patients
Authors: Penttilä  , Poulsen  , Hollingworth  , Holmström  
Published: March 2003

BackgroundTopical corticosteroids are the accepted medical adjunct to surgery in patients suffering from nasal polyposis. Fluticasone propionate (FP) is a potent, topically active corticosteroid which has been formulated as nasal drops specifically for the treatment of polyposis.ObjectivesTo evaluate dose‐related efficacy and tolerability of FP nasal drops (FPND) in the treatment of mild to moderate bilateral polyposis; in a double‐blind, placebo‐controlled, multicentre international study.MethodsAdult patients (n = 142) with bilateral nasal polyps were randomized to receive either FPND 400 μg once daily (o.d.), FPND 400 μg twice daily (b.i.d.) or placebo for 12 weeks. The majority then entered a further 12 week open period during which all patients received FPND 400 μg o.d. The primary efficacy endpoint was the physicians' visual assessment of polyp size. Secondary clinical endpoints were nasal blockage and overall rhinitis (0–3 scores), peak nasal inspiratory flow (PNIF), olfactory function tests, and requirement for polypectomy. The patients also kept twice daily records of symptom scores, peak nasal inspiratory flow (PNIF) and use of rescue medication.ResultsAt the end of the 12 week randomized treatment period, polyp size was reduced significantly by FPND 400 μg b.i.d. as compared with placebo (P = 0.006). Clinical assessments of nasal blockage and overall rhinitis showed significant improvements at several stages of treatment with both doses of FPND. Clinic PNIF was also improved significantly by both doses of FPND in comparison with placebo, and FPND 400 μg b.i.d. was significantly more effective than 400 μg o.d. (P = 0.045). Patient diary card scores supported the clinical assessments. Two patients on placebo required polypectomy and all treatments were well tolerated with a similar incidence of adverse events.ConclusionFPND 400 μg once or twice daily is an effective and well‐tolerated treatment for bilateral nasal polyposis.
Detection of immunoglobulin antibodies in the sera of patients using purified latex allergens
Authors: Kurup  , Yeang  , Sussman  , Bansal  , Beezhold  , Kelly  , Hoffman  , Williams  , Fink  
Published: March 2003

BackgroundLatex allergy is largely an occupational allergy due to sensitization to natural rubber latex allergens present in a number of health care and household products. Although several purified allergens are currently available for study, information on the usefulness of these purified, native or recombinant allergens in the demonstration of specific immunoglobulin (Ig) E in the sera of patients is lacking.ObjectiveTo evaluate the purified latex allergens and to demonstrate specific IgE antibody in the sera of health care workers and spina bifida patients with clinical latex allergy.MethodsTwo radioallergosorbent and an enzyme‐linked immunosorbent assay (ELISA) using latex proteins Hev b 1, 2, 3, 4, 6 and 7 along with two glove extracts and Malaysian nonammoniated latex (MNA) were evaluated to demonstrate IgE in the sera of health care workers and spina bifida with latex allergy and controls with no history of latex allergy.ResultsELISA using the purified latex allergens demonstrated specific IgE in 32–65% health care workers and 54–100% of spina bifida patients with latex allergy. The corresponding figures for RAST were 13–48 and 23–85 for RAST‐1 and 19–61 and 36–57 for RAST‐2. These results were comparable with the results obtained with glove extracts and crude rubber latex proteins.ConclusionsWhen used simultaneously, latex proteins Hev b 2 and Hev b 7 reacted significantly with specific serum IgE in 80% of health care workers and 92% of spina bifida patients with latex allergy by ELISA technique, while this combination gave lower positivity when the RASTs were used. By the addition of Hev b 3, specific IgE was detected in all spina bifida patients with latex allergy. Both RASTs failed to show specific IgE in the control subjects, while the ELISA showed significant latex‐specific IgE in 22% of controls.
Inhibition of CD23 processing correlates with inhibition of IL‐4‐stimulated IgE production in human PBL and hu‐PBL‐reconstituted SCID mice
Authors: Mayer  , Bolognese  , Al‐Mahdi  , Cook  , Flamberg  , Hansbury  , Khandekar  , Appelbaum  , Faller  , Marshall  
Published: March 2003

BackgroundCD23, the low affinity serum immunoglobulin E (IgE) receptor, is upregulated on B cells following interleukin (IL)‐4 stimulation and is concomitantly cleaved to generate soluble CD23 (sCD23) fragments with cytokine‐like activity.ObjectiveCompounds that selectively inhibit the proteolytic release of CD23 to generate sCD23 were assessed for their ability to inhibit IgE production in order to evaluate the contribution of sCD23 in the production of human IgE as well as the ability of such compounds to block IgE production.MethodsIgE production was measured in IL‐4‐stimulated human peripheral blood lymphocytes (PBL) and PBL‐reconstituted SCID mice in the presence of a broad‐spectrum matrix metalloprotease (MMP) inhibitor, a compound selective for inhibition of CD23 processing over MMPs and an anti‐CD23 mAb, MHM6.ResultsThe two compounds were equipotent in inhibiting IgE production without inhibition of IgG production by IL‐4/anti‐CD40‐stimulated PBL. Soluble CD23 release was also shown to precede IgE accumulation in the cell‐free medium. Addition of compound at later times other than day 0 in the 14 day assay resulted in progressively less inhibition of both IgE and sCD23, and exactly paralleled the effect of an anti‐CD23 mAb, MHM6 on IgE levels. Both compounds also inhibited the release of CD23 from human RPMI 8866 cells adoptively transferred i.p. to mice. Doses required for inhibition of CD23 correlated well with the doses required for inhibition of IgE production in IL‐4‐challenged hu‐PBL‐SCID mice. IgE was selectively inhibited over total IgG in the SCID mice as well.ConclusionsInhibition of CD23 processing alone is sufficient to inhibit IL‐4‐stimulated IgE production both in vitro and in vivo.
Cloning of the minor allergen Api g 4 profilin from celery (Apium graveolens) and its cross‐reactivity with birch pollen profilin Bet v 2
Authors: Scheurer  , Wangorsch  , Haustein  , Vieths  
Published: March 2003

BackgroundProfilin is a panallergen that is recognized by IgE from about 20% of birch pollen‐ and plant food‐allergic patients. A subgroup of celery‐allergic patients shows IgE‐reactivity with this minor allergen. To investigate the IgE‐binding potential and cross‐reactivity of celery profilin at the molecular level, this study was aimed at the cloning and immunological characterization of this allergen.ObjectivesCloning, expression and purification of profilin from celery tuber to characterize its immunological properties and its cross‐reactivity with birch pollen profilin.MethodsCloning of celery profilin was performed by polymerase chain reaction using degenerated primers and a 5′RACE method for the identification of the unknown 5′‐end of the cDNA. Expression was carried out in Escherichia coli BL21 (DE3) using a modified vector pET‐30a. The recombinant profilin was purified by affinity chromatography on poly l‐proline coupled to sepharose. Immunological characterization was performed by immunoblotting, EAST and IgE‐inhibition experiments.ResultsThe coding region of the cDNA of celery profilin was identified as a 399‐bp open reading frame, coding for a protein of 133 amino acids with a calculated molecular weight of 14.3 kDa. The deduced amino acid sequence of the corresponding protein showed high identity with other plant profilins (71–82%) recently described as allergens. Celery profilin was isolated as highly pure nonfusion protein. The IgE‐reactivity of celery profilin was similar to that of natural protein. Seven of 17 celery‐allergic patients tested presented specific IgE‐antibodies to the recombinant protein tested by immunoblotting. Inhibition experiments showed high cross‐reactivity of IgE with both profilins from celery and birch pollen. Moreover, the biological activity of recombinant celery profilin was demonstrated by a histamine release assay.ConclusionsCelery profilin is an important allergenic compound in celery and shows high homology to birch pollen profilin, Bet v 2. According to the revised IUIS allergen nomenclature, we suggest naming the celery profilin Api g 4. In addition to the cross‐reacting major allergens Api g 1 and Bet v 1, birch pollinosis and associated allergies to celery can therefore additionally be explained by the cross‐reactivity between homologous profilins. Moreover, recombinant Api g 4 may be used for target‐specific diagnosis and structural analyses.
Production and characterization of monoclonal antibodies to serine proteinase allergens in Penicillium and Aspergillus species
Authors: Lin W.‐L. , Chou H. , Tam M. F. , Huang M.‐H. , Han S.‐H. , Shen H.‐D. 
Published: March 2003

BackgroundAlkaline and/or vacuolar serine proteinases are major allergens in prevalent airborne Penicillium and Aspergillus species.ObjectiveThe object of this study is to generate and characterize monoclonal antibodies against these serine proteinase allergens.MethodsBALB/c mice were immunized individually with the Penicillium citrinum culture medium or the crude extract and culture medium preparations of Aspergillus fumigatus. Hybridoma cells that secrete monoclonal antibodies against serine proteinase allergens were selected by immunoblotting. Antigens in three different Penicillium (P. citrinum, P. notatum and P. oxalicum) and two different Aspergillus species (A. fumigatus, and A. flavus) recognized by these monoclonal antibodies were analysed by sodium dodecyl sulphate and two‐dimensional polyacrylamide gel electrophoresis immunoblotting and N‐terminal amino acid sequence analysis.ResultsFour (PCM8, PCM10, PCM16 and PCM39) and one (FUM20) monoclonal antibodies against serine proteinase allergens were generated after fusion of NS‐1 cells with spleen cells obtained from BALB/c mice immunized with antigens from P. citrinum and A. fumigatus, respectively. Immunoblotting results showed that PCM8 reacted with an alkaline serine proteinase allergen in P. citrinum and P. notatum. PCM10 and PCM39 reacted with the alkaline serine proteinase in two Penicillium (P. citrinum, P. notatum) and two Aspergillus species (A. fumigatus, and A. flavus) tested. PCM16 reacted with the alkaline serine proteinase allergen in P. citrinum, A. fumigatus and A. flavus but not with that in P. notatum. MoAb FUM20 reacted with the alkaline serine proteinase allergen in two Aspergillus species (A. fumigatus and A. flavus) but not with that in two different Penicillium species (P. citrinum, P. notatum) tested. Among these five monoclonal antibodies generated, only PCM39 and FUM20 can react with the vacuolar serine proteinase allergen in P. notatum, P. oxalicum and in A. fumigatus. The 35 kDa P. citrinum component that reacted with FUM20 has an N‐terminal amino acid sequence of DSPSVEKNAP.ConclusionFive monoclonal antibodies against different epitopes of the serine proteinase major allergens in prevalent Penicillium and Aspergillus species were generated in the present study. Antibodies obtained may be useful in the characterization and standardization of serine proteinase allergens in crude fungal extracts.
Early introduction of inhaled steroids in wheezing children presenting in primary care. A pilot study
Authors: Baxter‐Jones A. D. G. , Helms P. J. 
Published: March 2003

BackgroundRecurrent wheezing illness in childhood is a frequent reason for consultation in both primary and secondary care and whilst there is widespread support for the earlier introduction of inhaled corticosteroids(ICS) concerns remain about potential adverse systemic effects.ObjectiveWe sought to identify the feasibility and size of study required to assess the benefits, adverse effects and cost‐effectiveness of early introduction of ICS.MethodsWe entered 86 children (mean age 4.5 years) with at least two consecutive wheezing episodes into a pragmatic randomized controlled trial. All children were already being prescribed β2‐agonist and those allocated to ICS were prescribed twice daily beclomethasone dipropionate 200 μg or budesonide 200 μg. At entry and 3 and 6 months age appropriate lung function (FEV1, Tptef:Te), respiratory symptoms, quality of life of children and carers, growth and sexual maturation, bone mass and bone turnover, and health care costs were assessed.ResultsDrop out rate was 8% (7 of 86). No significant differences were found in FEV1 or Tptef:Te at 6 months. Respiratory symptoms and health‐related quality of life of children and carers improved equally in both groups. No significant differences were found in growth rates or in bone mass. In the ICS group markers of bone turnover (Pyd:Cr) fell (271 vs 237 nmol/mmol) in contrast to a rise in those on β2 alone (255 vs 319 nmol/mmol) P < 0.05. Combined health care costs were higher in the ICS group.ConclusionsThe low drop out rate provided evidence that the protocol was practical and acceptable. In order to confirm these results, and using the same study design, power estimates indicated that 260 children would need to be randomized.
Dietary factors associated with physician‐diagnosed asthma and allergic rhinitis in teenagers: analyses of the first Nutrition and Health Survey in Taiwan
Authors: Huang S.‐L. , Lin K.‐C. , Pan W.‐H. 
Published: March 2003

The occurrence of asthma and allergy are related to lifestyle factors, and dietary pattern may be one of the contributing factors.To examine the possible association between dietary intake and the prevalence of asthma and allergic rhinitis in teenagers.In a population‐based cross‐sectional survey, the relationship was sought between food frequency and physician‐diagnosed asthma and allergic rhinitis in 1166 adolescents aged 13–17.The prevalence was 4.0% for asthma and 12.4% for rhinitis. Living in an urbanized area was a significant predictor of asthma and rhinitis. In univariate analysis, higher frequencies of oily fish, butcher's meat, liver and deep‐fried foods were associated with asthma. Relevant food frequency variables were dichotomized at the 75th percentile for multivariate logistic regression analysis, which included adjustment for two levels of urbanization. Asthma was associated with intakes of liver (OR = 2.32, 95%CI 1.11–4.80), deep‐fried foods (OR = 2.13, 95%CI 1.06–4.30) and butcher's meat (OR 1.84, 95%CI 0.89–3.80). In a similar analysis, allergic rhinitis was associated with liver (OR = 1.67, 95%CI 1.06–2.63). No protective effect was demonstrated for any of the food items examined.Protein‐rich and fat‐rich foods of animal origin were associated with a higher prevalence of asthma in teenagers.